Posts Tagged ‘Cardiovascular disease’

Artificial Intelligence and Cardiovascular Disease

Reporter and Curator: Dr. Sudipta Saha, Ph.D.


Cardiology is a vast field that focuses on a large number of diseases specifically dealing with the heart, the circulatory system, and its functions. As such, similar symptomatologies and diagnostic features may be present in an individual, making it difficult for a doctor to easily isolate the actual heart-related problem. Consequently, the use of artificial intelligence aims to relieve doctors from this hurdle and extend better quality to patients. Results of screening tests such as echocardiograms, MRIs, or CT scans have long been proposed to be analyzed using more advanced techniques in the field of technology. As such, while artificial intelligence is not yet widely-used in clinical practice, it is seen as the future of healthcare.


The continuous development of the technological sector has enabled the industry to merge with medicine in order to create new integrated, reliable, and efficient methods of providing quality health care. One of the ongoing trends in cardiology at present is the proposed utilization of artificial intelligence (AI) in augmenting and extending the effectiveness of the cardiologist. This is because AI or machine-learning would allow for an accurate measure of patient functioning and diagnosis from the beginning up to the end of the therapeutic process. In particular, the use of artificial intelligence in cardiology aims to focus on research and development, clinical practice, and population health. Created to be an all-in-one mechanism in cardiac healthcare, AI technologies incorporate complex algorithms in determining relevant steps needed for a successful diagnosis and treatment. The role of artificial intelligence specifically extends to the identification of novel drug therapies, disease stratification or statistics, continuous remote monitoring and diagnostics, integration of multi-omic data, and extension of physician effectivity and efficiency.


Artificial intelligence – specifically a branch of it called machine learning – is being used in medicine to help with diagnosis. Computers might, for example, be better at interpreting heart scans. Computers can be ‘trained’ to make these predictions. This is done by feeding the computer information from hundreds or thousands of patients, plus instructions (an algorithm) on how to use that information. This information is heart scans, genetic and other test results, and how long each patient survived. These scans are in exquisite detail and the computer may be able to spot differences that are beyond human perception. It can also combine information from many different tests to give as accurate a picture as possible. The computer starts to work out which factors affected the patients’ outlook, so it can make predictions about other patients.


In current medical practice, doctors will use risk scores to make treatment decisions for their cardiac patients. These are based on a series of variables like weight, age and lifestyle. However, they do not always have the desired levels of accuracy. A particular example of the use of artificial examination in cardiology is the experimental study on heart disease patients, published in 2017. The researchers utilized cardiac MRI-based algorithms coupled with a 3D systolic cardiac motion pattern to accurately predict the health outcomes of patients with pulmonary hypertension. The experiment proved to be successful, with the technology being able to pick-up 30,000 points within the heart activity of 250 patients. With the success of the aforementioned study, as well as the promise of other researches on artificial intelligence, cardiology is seemingly moving towards a more technological practice.


One study was conducted in Finland where researchers enrolled 950 patients complaining of chest pain, who underwent the centre’s usual scanning protocol to check for coronary artery disease. Their outcomes were tracked for six years following their initial scans, over the course of which 24 of the patients had heart attacks and 49 died from all causes. The patients first underwent a coronary computed tomography angiography (CCTA) scan, which yielded 58 pieces of data on the presence of coronary plaque, vessel narrowing and calcification. Patients whose scans were suggestive of disease underwent a positron emission tomography (PET) scan which produced 17 variables on blood flow. Ten clinical variables were also obtained from medical records including sex, age, smoking status and diabetes. These 85 variables were then entered into an artificial intelligence (AI) programme called LogitBoost. The AI repeatedly analysed the imaging variables, and was able to learn how the imaging data interacted and identify the patterns which preceded death and heart attack with over 90% accuracy. The predictive performance using the ten clinical variables alone was modest, with an accuracy of 90%. When PET scan data was added, accuracy increased to 92.5%. The predictive performance increased significantly when CCTA scan data was added to clinical and PET data, with accuracy of 95.4%.


Another study findings showed that applying artificial intelligence (AI) to the electrocardiogram (ECG) enables early detection of left ventricular dysfunction and can identify individuals at increased risk for its development in the future. Asymptomatic left ventricular dysfunction (ALVD) is characterised by the presence of a weak heart pump with a risk of overt heart failure. It is present in three to six percent of the general population and is associated with reduced quality of life and longevity. However, it is treatable when found. Currently, there is no inexpensive, noninvasive, painless screening tool for ALVD available for diagnostic use. When tested on an independent set of 52,870 patients, the network model yielded values for the area under the curve, sensitivity, specificity, and accuracy of 0.93, 86.3 percent, 85.7 percent, and 85.7 percent, respectively. Furthermore, in patients without ventricular dysfunction, those with a positive AI screen were at four times the risk of developing future ventricular dysfunction compared with those with a negative screen.


In recent years, the analysis of big data database combined with computer deep learning has gradually played an important role in biomedical technology. For a large number of medical record data analysis, image analysis, single nucleotide polymorphism difference analysis, etc., all relevant research on the development and application of artificial intelligence can be observed extensively. For clinical indication, patients may receive a variety of cardiovascular routine examination and treatments, such as: cardiac ultrasound, multi-path ECG, cardiovascular and peripheral angiography, intravascular ultrasound and optical coherence tomography, electrical physiology, etc. By using artificial intelligence deep learning system, the investigators hope to not only improve the diagnostic rate and also gain more accurately predict the patient’s recovery, improve medical quality in the near future.


The primary issue about using artificial intelligence in cardiology, or in any field of medicine for that matter, is the ethical issues that it brings about. Physicians and healthcare professionals prior to their practice swear to the Hippocratic Oath—a promise to do their best for the welfare and betterment of their patients. Many physicians have argued that the use of artificial intelligence in medicine breaks the Hippocratic Oath since patients are technically left under the care of machines than of doctors. Furthermore, as machines may also malfunction, the safety of patients is also on the line at all times. As such, while medical practitioners see the promise of artificial technology, they are also heavily constricted about its use, safety, and appropriateness in medical practice.


Issues and challenges faced by technological innovations in cardiology are overpowered by current researches aiming to make artificial intelligence easily accessible and available for all. With that in mind, various projects are currently under study. For example, the use of wearable AI technology aims to develop a mechanism by which patients and doctors could easily access and monitor cardiac activity remotely. An ideal instrument for monitoring, wearable AI technology ensures real-time updates, monitoring, and evaluation. Another direction of cardiology in AI technology is the use of technology to record and validate empirical data to further analyze symptomatology, biomarkers, and treatment effectiveness. With AI technology, researchers in cardiology are aiming to simplify and expand the scope of knowledge on the field for better patient care and treatment outcomes.






























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Hypertriglyceridemia: Evaluation and Treatment Guideline

Reporter and Curator: Dr. Sudipta Saha, Ph.D.


Severe and very severe hypertriglyceridemia increase the risk for pancreatitis, whereas mild or moderate hypertriglyceridemia may be a risk factor for cardiovascular disease. Individuals found to have any elevation of fasting triglycerides should be evaluated for secondary causes of hyperlipidemia including endocrine conditions and medications. Patients with primary hypertriglyceridemia must be assessed for other cardiovascular risk factors, such as central obesity, hypertension, abnormalities of glucose metabolism, and liver dysfunction. The aim of this study was to develop clinical practice guidelines on hypertriglyceridemia.

The diagnosis of hypertriglyceridemia should be based on fasting levels, that mild and moderate hypertriglyceridemia (triglycerides of 150–999 mg/dl) be diagnosed to aid in the evaluation of cardiovascular risk, and that severe and very severe hypertriglyceridemia (triglycerides of >1000 mg/dl) be considered a risk for pancreatitis. The patients with hypertriglyceridemia must be evaluated for secondary causes of hyperlipidemia and that subjects with primary hypertriglyceridemia be evaluated for family history of dyslipidemia and cardiovascular disease.

The treatment goal in patients with moderate hypertriglyceridemia should be a non-high-density lipoprotein cholesterol level in agreement with National Cholesterol Education Program Adult Treatment Panel guidelines. The initial treatment should be lifestyle therapy; a combination of diet modification, physical activity and drug therapy may also be considered. In patients with severe or very severe hypertriglyceridemia, a fibrate can be used as a first-line agent for reduction of triglycerides in patients at risk for triglyceride-induced pancreatitis.

Three drug classes (fibrates, niacin, n-3 fatty acids) alone or in combination with statins may be considered as treatment options in patients with moderate to severe triglyceride levels. Statins are not be used as monotherapy for severe or very severe hypertriglyceridemia. However, statins may be useful for the treatment of moderate hypertriglyceridemia when indicated to modify cardiovascular risk.











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Changes in Levels of Sex Hormones and N-Terminal Pro–B-Type Natriuretic Peptide as Biomarker for Cardiovascular Diseases

Reporter and Curator: Dr. Sudipta Saha, Ph.D.


Considerable differences exist in the prevalence and manifestation of atherosclerotic cardiovascular disease (CVD) and heart failure (HF) between men and women. Premenopausal women have a lower risk of CVD and HF compared with men; however, this risk increases after menopause. Sex hormones, particularly androgens, are associated with CVD risk factors and events and have been postulated to mediate the observed sex differences in CVD.


B-type natriuretic peptides (BNPs) are secreted from cardiomyocytes in response to myocardial wall stress. BNP plays an important role in cardiovascular remodelling and volume homeostasis. It exerts numerous cardioprotective effects by promoting vasodilation, natriuresis, and ventricular relaxation and by antagonizing fibrosis and the effects of the renin-angiotensin-aldosterone system. Although the physiological role of BNP is cardioprotective, pathologically elevated N-terminal pro–BNP (NT-proBNP) levels are used clinically to indicate left ventricular hypertrophy, dysfunction, and myocardial ischemia. Higher NT-proBNP levels among individuals free of clinical CVD are associated with an increased risk of incident CVD, HF, and cardiovascular mortality.


BNP and NT-proBNP levels are higher in women than men in the general population. Several studies have proposed the use of sex- and age-specific reference ranges for BNP and NT-proBNP levels, in which reference limits are higher for women and older individuals. The etiology behind this sex difference has not been fully elucidated, but prior studies have demonstrated an association between sex hormones and NT-proBNP levels. Recent studies measuring endogenous sex hormones have suggested that androgens may play a larger role in BNP regulation by inhibiting its production.


Data were collected from a large, multiethnic community-based cohort of individuals free of CVD and HF at baseline to analyze both the cross-sectional and longitudinal associations between sex hormones [total testosterone (T), bioavailable T, freeT, dehydroepiandrosterone (DHEA), SHBG, and estradiol] and NT-proBNP, separately for women and men. It was found that a more androgenic pattern of sex hormones was independently associated with lower NT-proBNP levels in cross-sectional analyses in men and postmenopausal women.


This association may help explain sex differences in the distribution of NT-proBNP and may contribute to the NP deficiency in men relative to women. In longitudinal analyses, a more androgenic pattern of sex hormones was associated with a greater increase in NT-proBNP levels in both sexes, with a more robust association among women. This relationship may reflect a mechanism for the increased risk of CVD and HF seen in women after menopause.


Additional research is needed to further explore whether longitudinal changes in NT-proBNP levels seen in our study are correlated with longitudinal changes in sex hormones. The impact of menopause on changes in NT-proBNP levels over time should also be explored. Furthermore, future studies should aim to determine whether sex hormones directly play a role in biological pathways of BNP synthesis and clearance in a causal fashion. Lastly, the dual role of NTproBNP as both

  • a cardioprotective hormone and
  • a biomarker of CVD and HF, as well as
  • the role of sex hormones in delineating these processes,

should be further explored. This would provide a step toward improved clinical CVD risk stratification and prognostication based on

  • sex hormone and
  • NT-proBNP levels.














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Announcement 11AM- 5PM: Live Conference Coverage  from Mediterranean Diet and Lifestyle: A Symposium on Diet and Human Health @S.H.R.O. and Temple University October 19, 2018

Reporter: Stephen J. Williams, Ph.D.


 The Sbarro Health Research Organization, in collaboration with the Consulate General of Italy in Philadelphia will sponsor a symposium on the Mediterranean Diet and Human Health on October 19, 2018 at Temple University in Philadelphia, PA.  This symposium will discuss recent finding concerning the health benefits derived from a Mediterranean-style diet discussed by the leaders in this field of research.

Mediterranean Diet

The description of the Mediterranean Diet stems from the nutritionist Ancel Keys, who in 1945, in the wake of the US Fifth Army, landed in Southern Italy, where he observed one of the highest concentrations of centenarians in the world. He also noticed that cardiovascular diseases, widespread in the USA, were less frequent there. In particular, among the Southern Italians, the prevalence of “wellness” diseases such as hypertension and diabetes mellitus, was particularly associated with fat consumption, suggesting that the main factor responsible for the observations was the type of diet traditionally consumed among people facing the Mediterranean Sea, which is low in animal fat, as opposed to the Anglo-Saxon diet. The link between serum cholesterol and coronary heart disease mortality was subsequently demonstrated by the Seven Countries Study. Later, the concept of Mediterranean Diet was extended to a diet rich in fruits, vegetables, legumes, whole grains, fish and olive oil as the main source of lipid, shared among people living in Spain, Greece, Southern Italy and other countries facing the Mediterranean basin …

Prof. Antonino De Lorenzo, MD, PhD.



The Symposium will be held at:

Biolife Science Building, Room 234

Temple University, 1900 North 12th street

Philadelphia, PA 19122


For further information, please contact:

Ms. Marinela Dedaj – Sbarro Institute,  Office #: 215-204-9521


11:00 Welcome

Prof. Antonio Giordano, MD, PhD.

Director and President of the Sbarro Health Research Organization, College of Science and Technology, Temple University



Fucsia Nissoli Fitzgerald

Deputy elected in the Foreign Circumscription – North and Central America Division


Consul General, Honorable Pier Attinio Forlano

General Consul of Italy in Philadelphia


11:30 The Impact of Environment and Life Style in Human Disease

Prof. Antonio Giordano MD, PhD.


12.00 The Italian Mediterranean Diet as a Model of Identity of a People with a Universal Good to Safeguard Health?

Prof. Antonino De Lorenzo, MD, PhD.

Director of the School of Specialization in Clinical Nutrition, University of Rome “Tor Vergata”


12:30 Environment and Health

Dr. Iris Maria Forte, PhD.

National Cancer Institute “Pascale” Foundation | IRCCS · Department of Research, Naples, Italy


13:00 Lunch


2:30 Mediterranean Diet, Intangible Heritage and Sustainable Tourism?

Prof. Fabio Parasecoli, PhD.

Nutrition and Food Department, New York University


3.00 Italy as a Case Study: Increasing Students’ Level of Awareness of the Historical, Cultural, Political and Culinary Significance of Food

Prof. Lisa Sasson

Nutrition and Food Department, New York University


3:30 Italian Migration and Global Diaspora

Dr. Vincenzo Milione, PhD

Director of Demographics Studies, Calandra Institute, City University of New York


4:00 Pasta Arte: New Model of Circular Agricultural Economy: When an Innovated Tradition Takes Care of You and of the Environment

Dr. Massimo Borrelli

CEO and Founder of Arte


4:15 Conclusions

Prof. Antonio Giordano, MD, PhD.


Coordinator of the Symposium, Dr. Alessandra Moia, PhD.


Prof. Antonio Giordano, MD, PhD.

Professor of Molecular Biology at Temple University in Philadelphia, PA where he is also Director of the Sbarro Institute for Cancer Research and Molecular Medicine. He is also Professor of Pathology at the University of Siena, Italy. He has published over 500 articles, received over 40 awards for his contributions to cancer research and is the holder of 17 patents.


Prof. Antonino De Lorenzo, MD, PhD.

Full Professor of Human Nutrition and Director of the Specialization School in Food Science at the University of Rome “Tor Vergata”. He is the Coordinator of the Specialization Schools in Food Science at the National University Council and Coordinator of the PhD. School of “Applied Medical-Surgical Sciences” Director of UOSD “Service of Clinical Nutrition, Parenteral Therapy and Anorexia”. He also serves as President of “Istituto Nazionale per la Dieta Mediterranea e la Nutrigenomica”.


Dr. Iris Maria Forte, PhD.

Iris Maria Forte is an oncology researcher of INT G. Pascale Foundation of Naples, Italy. She majored in Medical Biotechnology at the “Federico II” University of Naples, earned a PhD. in “Oncology and Genetics” at the University of Siena in 2012 and a Master of II level in “Environment and Cancer” in 2014. Iris Maria Forte has worked with Antonio Giordano’s group since 2008 and her research interests include both molecular and translational cancer research. She published 21 articles mostly focused in understanding the molecular basis of human cancer. She worked on different kinds of human solid tumors but her research principally focused on pleural mesothelioma and on cell cycle deregulation in cancer.


Prof. Fabio Parasecoli, PhD.

Professor in the Department of Nutrition and Food Studies. He has a Doctorate in Agricultural Sciences (Dr.sc.agr.) from Hohenheim University, Stuttgart (Germany), MA in Political Sciences from the Istituto Universitario Orientale, Naples (Italy), BA/MA in Modern Foreign Languages and Literature from the Università La Sapienza, Rome (Italy). His research explores the intersections among food, media, and politics. His most recent projects focus on Food Design and the synergies between Food Studies and design.


Prof. Lisa Sasson, MS

Dietetic Internship Director and a Clinical Associate Professor in the department. She has interests in dietetic education, weight and behavior management, and problem-based learning. She also is a private practice nutritionist with a focus on weight management. She serves as co-director of the Food, Nutrition and Culture program in Florence Italy, the New York State Dietetic Association and the Greater New York Dietetic Association (past president and treasurer).


Dr. Vincenzo Milione, PhD.

Director of Demographic Studies for The John D. Calandra Italian American Institute, Queens College, City University of New York. He has conducted social science research on Italian Americans. His research has included the educational and occupational achievements; Italian language studies at the elementary and secondary levels, high school non-completion rates; negative media portrayals of ethnic populations including migration studies and global diaspora.


Dr. Massimo Borrelli

Agricultural entrepreneur, Manager of the Italian Consortium for Biogas (CIB) and delegate for the Bioeconomy National Department of Confagricoltura. He developed A.R.T.E based on a model of agricultural circular economy, beginning and ending in the ground. He constructed the first biogas plant in the territory creating a new way to make agriculture, investing in research and development, experimentation and most of all, in people. In a few short years, he succeeded to close the production chain producing goods characterized by their high quality and usage of renewable energy.


Dr. Alessandra Moia, PhD.

Vice-President for Institutional and International Relations of the Istituto Nazionale per la Dieta Mediterranea e la Nutrigenomica (I.N.D.I.M.). Has managed relations with the academic institutions to increase awareness and develops projects for the diffusion of the Mediterranean Diet. She served as Director of Finance for the National Institute of Nutrition, for the Ministry of Agriculture and Forestry.


About the Sbarro Health Research Organization

The Sbarro Health Research Organization (SHRO) is non-profit charity committed to funding excellence in basic genetic research to cure and diagnose cancer, cardiovascular diseases, diabetes and other chronic illnesses and to foster the training of young doctors in a spirit of professionalism and humanism. To learn more about the SHRO please visit www.shro.org

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Ralph’s Story: An Entertainer at Heart

Patient was diagnosed with heart disease and pulmonary hypertension in January 2016 and had a triple-bypass operation at age 69. Interview was conducted six months post-surgery.

Author: Gail S. Thornton, M.A.

Co-Editor: The VOICES of Patients, HealthCare Providers, Caregivers and Families: Personal Experience with Critical Care and Invasive Medical Procedures


Evergreen, Colorado, an idyllic, peaceful community with an elevation of 8,000 feet west of Denver, offers its residents and visitors a beautiful place for arts and culture, summer and winter sporting activities, and scenic beauty. In fact, Ralph Nichols has lived in the town for more than 20 years.

“This past September [2015] was, particularly, challenging for me, where winter begins quite early for us. It became increasingly painful and difficult to breathe in the freezing temperatures. It seemed that my lungs were inflamed and I couldn’t even stand the cold weather. I thought it might be the beginning of a bad cold, and I wasn’t overly concerned that there was anything terribly wrong.”

At that time, Ralph went to his family physician who performed the usual routine examination with no significant results.

“Many years ago, I developed a mild case of scleroderma, a chronic connective tissue disease. I thought that perhaps my symptoms were the result of some type of inflammation in my body that could be managed with prescription medications.”

Scleroderma is known as an autoimmune disease, which adds an inappropriate amount of collagen to various parts of the body, such as the joints, skin, and later stages, various organs, such as the lungs, in Ralph’s case. Scleroderma can cause the organs to shut down and, eventually, cause death.

“I never let this condition stop me from doing anything as it is life-long condition. It was always something I had to tolerate and work through.”



Image SOURCE: Photographs courtesy of Ralph Nichols and Gabriela Contreras.  Top left: Ralph today. Top right: Ralph recovering one month after surgery. Bottom left and center: Ralph with his medical team. Bottom right: Ralph in rehabilitation center.

Over the brutal Colorado winter, Ralph’s symptoms were getting worse. He had no idea that his life would dramatically change over the next few months. He went to see his family physician again. During this physical examination, Ralph was referred to pulmonary and cardiovascular specialists for a routine electrocardiogram, echocardiogram and stress test in order to further diagnose his symptoms. He had always been relatively healthy and fit and never been seriously ill or hospitalized.

“On the outside, Ralph was the picture of good health,” said his wife, Gabriela. “On the inside, his body was telling him that something was wrong.”

Three months later in December 2015, Ralph met with Dr. Alexandra Smart, a pulmonologist, who ordered a chest x-ray and other diagnostic tests, including a right heart catheterization. At that point, Ralph’s medical team grew. It was then determined that Ralph needed to see other cardiovascular specialists and undergo more tests. In January 2016, he met with Dr. Sameer Mehta, cardiologist at Cardiac & Thoracic Surgery Associates, in Lakewood, Colorado, who reviewed his tests to date, listened to Ralph’s symptoms, and told him he needed both a right and left heart cardiac catheterization.

 “They gave me sedation for the catheterization procedure and went through my neck with a camera to see what was going on with my lungs and heart. We were all singing together on the way to the operating room. During the procedure, my cardiologist found more than he had anticipated.”

The result was not good. Ralph had major blockages in two main arteries that supply blood to his heart muscle compounded by the fact that his lungs were affected by scleroderma.

“The catheterization was alarming. It showed that my arteries were in bad shape. They were both clogged with atherosclerotic plaque; one of them was 99 percent blocked and the other was 85 percent blocked.”

His cardiologist believed that the blockages would not respond to medications quickly or a stent.

“Even though my father had major heart disease and died two years later of cancer at the age of 56, I thought that I would be immune to this particular experience. After all, I was in good health, exercised regularly, lived a reasonable lifestyle and had a great diet.”

 Preparing for Life-Saving and Life-Changing Surgery

Unfortunately, surgery was the next step. Ralph was referred to Dr. Mehta’s colleague, Dr. Patrick D. Rudersdorf, cardiothoracic surgeon at Cardiac & Thoracic Surgery Associates.

“I didn’t leave the hospital that day as expected. Instead, I got a visit from Dr. Rudersdorf and couldn’t believe what he was telling me. My only chance to live was having triple bypass surgery which needed to be done immediately. The doctor met with me that same day to explain the procedure, answer my questions and talk through the details of the rehabilitation period after the surgery.”

Dr. Rudersdorf reassured Ralph that he was doing the right thing and calmed my fears.

“He said that I needed this life-saving surgery because I was at high risk for having a major heart attack. I was shocked, at first, at the thought of the intensity of surgery on my body. It’s a situation that no one likes to be in, but I had to make a decision about alleviating the ongoing pain and pressure in my chest along with shortness of breath due to diseased heart arteries. Coronary bypass surgery was my answer to feeling better — and it essentially gave me my life back.”

Dr. Rudersdorf moved his previously planned morning surgery to another day to accommodate me first thing in the morning. Ralph underwent triple bypass surgery at St. Anthony Hospital in Lakewood, Colorado. The procedure was complex and took eight hours. He was in the hospital for a total of 31 days.

“It was an ordeal that I thought I’d never have to experience. I had no time to call anyone, or time to even contemplate life and death…or even being scared.  My wife Gabriela spent the entire time in the hospital, supported by our dearest friends, Norma Delaney and Garret Annofsky, in addition to keeping family and friends in other parts of the United States and Mexico updated as well. Once the surgery was over, the medical team woke me up and said the procedure was successful, but I was far from being out of the woods.”

Ralph had some complications because of a condition called pulmonary hypertension, a type of high blood pressure that affects the arteries in the lungs and the right side of the heart. According to the Mayo Clinic’s web site, in one form of pulmonary hypertension, tiny arteries in the lungs, called pulmonary arterioles, and capillaries become narrowed, blocked or destroyed. This makes it harder for blood to flow through the lungs, and raises pressure within the lungs’ arteries. As the pressure builds, the heart’s lower right chamber (right ventricle) must work harder to pump blood through the lungs, eventually causing the heart muscle to weaken and fail. http://www.mayoclinic.org/diseases-conditions/pulmonary-hypertension/home/ovc-20197480

“The pulmonary hypertension limited some of the medications that the doctors would have used during my recovery. It was a tough few days for me in intensive care, hooked up to about 18 monitors. The medical team had to stop and re-start my heart four different times because of atrial fibrillation — finally getting both parts of the heart to dance together in the same rhythm.”

Ralph’s heart was beating abnormally fast and irregular and not functioning the way it should. The doctors restore regular rhythm to the heart by sending an electrical shock to the heart, which is called electrical cardioversion or chemically using antiarrhythmia medications, which is called pharmacologic or chemical cardioversion.

“The doctors shocked my heart first chemically with medications when I was awake. This procedure was the scariest. I was sitting up in bed and felt my heart stop, then the medical team flushed the medication out with saline in order to restart my heart. That procedure was not successful, so that is why the doctors had to shock my heart three more times electrically.

“The reason the doctors stopped my heart was to correct the atrial fibrillation and to get my heart into regular sinus rhythm, which is a wave mode of the heart where everything is synchronized. The doctors did not want me to continue to experience atrial fibrillation because if continued, I would not be able to regain my strength.”

Ralph was finally moved from intensive care to intermediate care after five days and the medical team kept him in intermediate care another 12 days until his heart and lungs got stronger.

“From there, I didn’t go home but instead went to Evergreen Life Center for rehabilitation for two weeks to learn how to walk, climb stairs so that I could access my home on my own, and develop my strength again. The rehab team would let me leave only after making sure I had oxygen in my home.”

After that, Ralph started another phase of his rehabilitation at St. Anthony Cardiac Rehabilitation and Wellness Center. For the next three months, he took part in cardiac rehabilitation three days a week. He passed that with flying colors. Now, he is in another phase of rehabilitation, building his lung capacity two days a week.

Ralph didn’t have the means or even the will to communicate with friends during this tumultuous time, except Gabriela and several close friends who were always at the hospital and rehabilitation center who gave him the strength to continue.

“I finally returned home after many weeks with an enormous feeling of gratitude for each and every one of my friends, as well as the St. Anthony’s hospital team of doctors, nurses, and therapists, who supported me and Gabriela during this exceptional adventure that has certainly changed my life.”

Surely, this experience has been a life-changing experience for Ralph.

 Coronary Artery Bypass Facts

 Coronary artery bypass grafting (CABG, often pronounced “cabbage”) is a surgical treatment for blocked coronary arteries. Coronary arteries supply blood to the heart muscle and when blockages in these arteries form, chest pain, shortness of breath and heart attacks can occur. Catheter procedures performed by interventional cardiologists address the blockages themselves with stents. Coronary bypass surgery performed by cardiac surgeons reroutes the blood around the blockages to supply better blood supply to the heart muscle and is a better treatment option, although more invasive, for certain patients and more durable for most patients.


Life for Ralph Today

Today, Ralph is regaining his strength both in mind and body. He visits the cardiovascular and pulmonary rehabilitation center three times a week for the past few months and walks on their treadmill, lifts weights and pedals the bicycle for one hour, supervised by the therapists. He also sees his medical team for regular check-ups every month, eats healthier with no fat and no salt, and takes a cocktail of medicines daily for his heart and lungs, including amiodarone, furosemide, pitavastatin, and aspirin.

“Almost six months after my surgery, although I am not in the best shape of my life, however, I am in the best spiritual place than ever before. This is a huge milestone for me. I continue to improve my strength, which will make my heart more resilient. There is nothing that I can’t do now, and I am doing everything I can to experience a normal life as far as work and regaining my strength. I find it necessary to move to a warmer climate and lower altitude in order to continue to improve.”

Ralph also is the former lead singer of The Letterman and The Sandpipers, two American easy-listening bands during the 1960-70-80s. He is an entertainer at heart with over 3,000 professional appearances to his credit. He has been performing and recording for over 50 years, traveled the world extensively and performed before members of the Vatican with Pope Pius XII and Royalty with Prince Rainier and Princess Grace Kelly, as well as notables such as Frank and Nancy Sinatra, Tony Bennett, Ronald Reagan, Merv Griffin, Danny Thomas, Shirley Bassey, Rosalind Russell and Bob Hope.

Ralph and his vocal group were dubbed by Billboard Magazine as “the greatest romantic vocal group of all time.” He is also a member of the Vocal Group Hall of Fame, a prestigious honor. He is a true legend as his group has sold more than 20 million recordings, performed live thousands of times, and whose recording of the song “Love” was left by NASA astronauts in a time capsule on the moon.

“I enjoy each and every day and appreciate all that life has to offer.”

Ralph’s next step is to get back to singing and his solo entertainment business, which he holds dear to his heart. That should be a task that he can easily accomplish.


Editor’s note:

We would like to thank Gabriela Contreras, a global communications consultant and patient advocate, for the tremendous help and support that she provided in scheduling time to talk with Ralph Nichols.

Ralph Nichols provided his permission to publish this interview on July 30, 2016.







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A Patient’s Perspective: On Open Heart Surgery from Diagnosis and Intervention to Recovery


No evidence to change current transfusion practices for adults undergoing complex cardiac surgery: RECESS evaluated 1,098 cardiac surgery patients received red blood cell units stored for short or long periods



ACC/AHA Guidelines for Coronary Artery Bypass Graft Surgery


On Devices and On Algorithms: Arrhythmia after Cardiac SurgeryPrediction and ECG Prediction of Paroxysmal Atrial Fibrillation Onset



Editor’s note:

I wish to encourage the e-Reader of this Interview to consider reading and comparing the experiences of other Open Heart Surgery Patients, voicing their private-life episodes in the ER that are included in this volume.

I also wish to encourage the e-Reader to consider, if interested, reviewing additional e-Books on Cardiovascular Diseases from the same Publisher, Leaders in Pharmaceutical Business Intelligence (LPBI) Group, on Amazon.com.

  •  Perspectives on Nitric Oxide in Disease Mechanisms, on Amazon since 6/2/12013


  • Cardiovascular, Volume Two: Cardiovascular Original Research: Cases in Methodology Design for Content Co-Curation, on Amazon since 11/30/2015


  • Cardiovascular Diseases, Volume Three: Etiologies of Cardiovascular Diseases: Epigenetics, Genetics and Genomics, on Amazon since 11/29/2015


  • Cardiovascular Diseases, Volume Four: Regenerative and Translational Medicine: The Therapeutics Promise for Cardiovascular Diseases, on Amazon since 12/26/2015




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Praluent FDA Approved

Larry H. Bernstein, MD, FCAP, Curator



PRALUENT® (alirocumab) is now approved for additional LDL-C lowering on top of maximally tolerated statin therapy in patients with HeFH or clinical ASCVD1
PRALUENT is a PCSK9 (Proprotein Convertase Subtilisin/Kexin Type 9) inhibitor antibody indicated as adjunct to diet and maximally tolerated statin therapy for the treatment of adults with heterozygous familial hypercholesterolemia or clinical atherosclerotic cardiovascular disease, who require additional lowering of LDL-C.

The effect of PRALUENT on cardiovascular morbidity and mortality has not been determined.

The recommended starting dose of PRALUENT is 75 mg administered subcutaneously once every 2 weeks, since the majority of patients achieve sufficient LDL-C reduction with this dosage. If the LDL-C response is inadequate, the dosage may be increased to the maximum dosage of 150 mg administered every 2 weeks.

Measure LDL-C levels within 4 to 8 weeks of initiating or titrating PRALUENT to assess response and adjust the dose, if needed.

PRALUENT is a human monoclonal antibody that binds to PCSK91
PRALUENT efficacy was investigated in 5 double-blind, placebo-controlled trials with 3499 patients enrolled: 36% with HeFH and 54% non-FH with clinical ASCVD.
All patients were receiving a maximally tolerated dose of statin with or without other lipid-modifying therapies
3 studies used an initial dose of 75 mg Q2W as part of an up-titration regimen with criteria-based up-titration to 150 mg Q2W at week 12 for patients who did not achieve their prespecified target LDL-C at week 8
2 studies with 150 mg Q2W dose only
Clinical ASCVD is defined in the ACC/AHA guidelines2 as acute coronary syndromes or a history of any of the following: myocardial infarction, stable or unstable angina, coronary or other arterial revascularization, transient ischemic attack or stroke, or peripheral arterial disease presumed to be of atherosclerotic origin.
All studies met their primary efficacy endpoint measured at week 241
All trials were at least 52 weeks in duration with the primary efficacy endpoint measured at week 24 (mean percent change in LDL-C from baseline)
The first and only FDA-approved PCSK9 inhibitor with 2 doses that allows you to adjust the dose based on your patients’ LDL-C lowering needs1
MyPRALUENT™: Comprehensive support for you and your patients
MyPRALUENT is designed to help meet your needs and your patients’ needs
Speak with a MyPRALUENT Care Specialist at 1-844-PRALUENT (1-844-772-5836), option 1
PRALUENT is contraindicated in patients with a history of a serious hypersensitivity reaction to PRALUENT. Reactions have included hypersensitivity vasculitis and hypersensitivity reactions requiring hospitalization.

Hypersensitivity reactions (e.g., pruritus, rash, urticaria), including some serious events (e.g., hypersensitivity vasculitis and hypersensitivity reactions requiring hospitalization), have been reported with PRALUENT treatment. If signs or symptoms of serious allergic reactions occur, discontinue treatment with PRALUENT, treat according to the standard of care, and monitor until signs and symptoms resolve.

The most commonly occurring adverse reactions (≥5% of patients treated with PRALUENT and occurring more frequently than with placebo) are nasopharyngitis, injection site reactions, and influenza.

Local injection site reactions including erythema/redness, itching, swelling, and pain/tenderness were reported more frequently in patients treated with PRALUENT (7.2% versus 5.1% for PRALUENT and placebo, respectively). Few patients discontinued treatment because of these reactions (0.2% versus 0.4% for PRALUENT and placebo, respectively), but patients receiving PRALUENT had a greater number of injection site reactions, had more reports of associated symptoms, and had reactions of longer average duration than patients receiving placebo.

Neurocognitive events were reported in 0.8% of patients treated with PRALUENT and 0.7% of patients treated with placebo. Confusion or memory impairment were reported more frequently by those treated with PRALUENT (0.2% for each) than in those treated with placebo (<0.1% for each).

Liver-related disorders (primarily related to abnormalities in liver enzymes) were reported in 2.5% of patients treated with PRALUENT and 1.8% of patients treated with placebo, leading to treatment discontinuation in 0.4% and 0.2% of patients, respectively. Increases in serum transaminases to greater than 3 times the upper limit of normal occurred in 1.7% of patients treated with PRALUENT and 1.4% of patients treated with placebo.

The most common adverse reactions leading to treatment discontinuation in patients treated with PRALUENT were allergic reactions (0.6% versus 0.2% for PRALUENT and placebo, respectively) and elevated liver enzymes (0.3% versus <0.1%).

PRALUENT is a human monoclonal antibody. As with all therapeutic proteins, there is a potential for immunogenicity with PRALUENT.

Please see full Prescribing Information.


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Troponin T elevation has 20-fold increased ESRD rate

Larry H. Bernstein, MD, FCAP, Curator


Troponin T Predicts End-Stage Renal Disease, Death

NEPHROLOGY 10.23.2015

Salynn Boyles


The presence of elevated cardiac troponin T in the blood was found to be an early independent predictor of end-stage renal disease and death in both African Americans and whites with hypertension or a family history of high blood pressure, researchers said.

Cardiac troponin T (cTnT) testing is routinely used to diagnose myocardial infarction or assess heart muscle damage in this setting, and several studies suggest that minimally elevated cTnT levels are associated with greater all-cause death risk in older people and in patients with chronic kidney failure.

The newly published study is the first to suggest a role for cTnT as a predictor of end-stage renal disease in patients with hypertension, nephrologistLaTonya Hickson, MD, and colleagues from the Mayo Clinic in Rochester, Minn., wrote in the journal Mayo Clinic Proceedings.

At 10-year follow-up, the estimated cumulative incidence of end-stage renal disease (ESRD) was 27.4% among study participants with abnormal cTnT levels (0.01 ng/mL or higher), compared to 1.3% among participants with lower cTnT levels.

“As patient populations grow older with increasing multimorbidity, identifying those at the highest risk of death or end-stage renal disease could improve patient management strategies,” the researchers wrote.

In an interview with MedPage Today, Hickson noted that cTnT predicted long-term ESRD and death independently of traditional risk factors for cardiovascular disease or end-stage renal disease

Several other recent studies have shown that a highly sensitive assay for cTnT, not yet clinically available in the U.S., can predict death and cardiovascular disease in community-dwelling older people.

For example, an analysis of 4,221 older people participating in the NHLBI’s Cardiovascular Health Study found that elevated levels of cTnT measured with such an assay were associated with a higher risk for cardiovascular death and heart failureindependent of other biomarkers.

Hickson said the idea for examining cTnT as a potential predictor of ESRD stemmed from earlier research by the Mayo team showing an increased risk for death associated with the highest cTnT levels among patients with kidney failure undergoing transplants.

“We now use this marker at Mayo to assess these patients,” she said.

The study included just over 3,000 participants in the National Heart, Lung, and Blood Institute’s GENOA study, originally designed to identify genetic determinants of hypertension in various racial groups.

Just under half (45%) of the participants were white residents of the Rochester, Minn., area and the rest were African American residents of Jackson, Miss. At baseline, a total of 71.3% were hypertensive and 32.5% had evidence of abnormal kidney function (glomerular filtration rate of less than 60 mL/min per 1.73 m2). About half (51.6%) had high sensitivity C-reactive protein levels greater than 3 mg/L.

Just 2.2% (66 of 3,050) had abnormal cTnT levels of 0.01 ng/mL or higher.

In addition to the 20-fold increase seen in risk for ESRD among those with abnormal cTnT, their estimated cumulative incidence of death at 10 years was 47%, compared to 7.3% among those with normal cTnT.

“Abnormal cTnT levels were strongly associated with ESRD and death,” the researchers wrote. “This effect was attenuated but was still highly significant after adjustment for demographic characteristics, estimated glomerular filtration rate, and traditional risk factors for ESRD,” they wrote, with an adjusted hazard ratio of 2.81 (95% CI 1.3-5.9) for ESRD and 3.46 (95% CI 2.3-5.1) for death.

Potential study limitations cited by the researchers included the lack of baseline measurement of urine protein excretion rates in addition to electrocardiogram and echocardiogram studies, “which may have provided insight into the presence of left ventricular hypertrophy, a condition previously associated with abnormal cTnT levels.”

They also noted that limiting the cohort to people with hypertension or belonging to hypertensive families may impact the study’s generalizability.

The researchers concluded that as patient populations grow older and develop more health issues, identifying those with the greatest risk for ESRD will become more important.

“Unfortunately, abnormal cTnT levels, measured with the standard assay, are relatively uncommon and thus do not improve risk prediction enough to support routine use,” they wrote. “Further study is needed to determine whether there is a particular patient group in which cTnT screening would meaningfully improve discrimination between the low- and high-risk patients for these sequelae.”

Funding for this research was provided by the Mayo Foundation, the National Institues of Health, and a Mary Kathryn and Michael B. Panitch Career Development Award.

Co-author Allan S. Jaffe reported receiving consulting fees from “most of the major diagnostic companies.” The other researchers reported no relevant relationships with industry.

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Writer and Curator: Larry H. Bernstein, MD, FCAP



The previous discussion concerned genomics, metabolomics, and cancer. The discussion that follows is concerned with the expanding filed of proteomics, which has implication for disease discovery, pharmaceutical targeting, and diagnostics.

The human proteome – a scientific opportunity for transforming diagnostics, therapeutics, and healthcare

Marc Vidal, Daniel W Chan, Mark Gerstein, Matthias Mann, Gilbert S Omenn, et al.
Clinical Proteomics 2012, 9:6  http://www.clinicalproteomicsjournal.com/content/9/1/6

A National Institutes of Health (NIH) workshop was convened in Bethesda, MD on September 26–27, 2011, with representative scientific leaders in the field of proteomics and its applications to clinical settings. The main purpose of this workshop was to articulate ways in which the biomedical research community can capitalize on recent technology advances and synergize with ongoing efforts to advance the field of human proteomics. This executive summary and the following full report describe the main discussions and outcomes of the workshop.

Proteomics Pioneer Award 2013: Professor Amos Bairoch, University of Geneva, Switzerland

Eupa Open Proteomics 2 (2014) 34  http://dx.doi.org/10.1016/j.euprot.2013.12.002

Amos Bairoch has always been fascinated by computer science, genetics and biochemistry. His fi rst project, as a PhD student, was the development of PC/Gene, a MS-DOS based software package for the analysis of protein and nucleotide sequences. While working on this project, he realized that there was no single resource for protein sequences, and started to develop the first annotated protein sequence database, which became Swiss-Prot and was first released in July 1986. In 1988, he created PROSITE, a database of protein families and domains, and a little later ENZYME, an enzyme nomenclature database.

Amos Bairoch led the Swiss-Prot group from its creation in 1988 until 2009. During this period, Swiss-Prot became the primary protein sequence resource in the world and has been a key research instrument for both bioinformaticians and laboratory-based scientists, particularly in the field of proteomics.

Since 2009, Amos Bairoch’s group is developing neXtProt, a knowledgebase
specifically dedicated to human proteins.neXtProt has been chosen as the reference protein database for the HUPO Human Proteome Projects.

For his major contributions in the field of proteomic databases, Amos Bairoch received the Friedrich Miescher Award from the Swiss Society of Biochemistry in 1993, the Helmut Horten Foundation Incentive Award in 1995, the Pehr Edman award and the European Latsis Prize in 2004, the Otto Naegeli prize in 2010, and the HUPO Distinguished Achievement Award in Proteomic Sciences in 2011.

National Heart, Lung, and Blood Institute Clinical Proteomics Working Group Report

CB Granger, JE Van Eyk, SC Mockrin and N. Leigh Anderson
Circulation. 2004;109:1697-1703

The National Heart, Lung, and Blood Institute (NHLBI) Clinical Proteomics Working Group was charged with identifying opportunities and challenges in clinical proteomics and using these as a basis for recommendations aimed at directly improving patient care. The group included representatives of clinical and translational research, proteomic technologies, laboratory medicine, bioinformatics, and 2 of the NHLBI Proteomics Centers, which form part of a program focused on innovative technology development. This report represents the results from a one-and-a-half-day meeting on May 8 and 9, 2003. For the purposes of this report, clinical proteomics is defined as the systematic, comprehensive, large-scale identification of protein patterns (“fingerprints”) of disease and the application of this knowledge to improve patient care and public health through better assessment of disease susceptibility, prevention of disease, selection of therapy for the individual, and monitoring of treatment response.

The -omics era: Proteomics and lipidomics in vascular research

Athanasios Didangelos, Christin Stegemann, Manuel Mayr
Atherosclerosis 221 (2012) 12– 17

The retention of proatherogenic low-density lipoprotein (LDL) particles on the subendothelial extracellular matrix (ECM) is a hallmark of atherosclerosis. Apolipoprotein B (apoB)-containing lipoprotein particles are trapped in the arterial intima by proteoglycans in atherosclerosis-prone areas and eventually become modified, commonly by aggregation and oxidation. The initial accumulation of proatherogenic lipoproteins initiates an inflammatory response, which results in the release of proteolytic enzymes and induces the dedifferentiation of vascular smooth muscle cells (SMCs) resulting in alterations of their matrix producing properties. The precise mechanisms responsible for the accumulation of certain matrix components and subsequent lipoprotein retention on the vessel wall are not fully elucidated. Undoubtedly, ECM remodeling contributes to the formation of atherosclerotic lesions and the lipid composition of apolipoproteins influences their binding properties to the matrix. An unbiased discovery approach, which is not limited to known molecules of presumed importance, will be invaluable for the identification of novel, previously unknown mediators of disease. Although descriptive, the detailed examination of atherosclerotic plaques using advanced proteomics and lipidomics techniques can generate novel insights and form the basis for further mechanistic investigations.

The Revolution in Proteomics Ionization –
CaptiveSpray nanoBooster™
Bruker, LC-MS Source

Bruker CaptiveSpray principle:

Stable and robust nanoflow LC/MS is still a challenge in proteomics analysis. The Bruker CaptiveSpray source is a revolutionary ion source with a patented design that provides provides easy operation just as simple normal flow electrospray.

CaptiveSpray delivers nanospray sensitivity, resists plugging, and provides reproducible uninterrupted flow for even the most complex proteomics samples.

CaptiveSpray nanoBooster brings your MS to the next performance level and provides even higher flexibility.

  • Boost nanoflow sensitivity
    • Push up ID rates
    • Enabling Glycoanalysis
    • Supercharging capability

CaptiveSpray provides a vortex gas that sweeps around the emitter spray tip to desolvate and to focus the Taylor cone into the MS inlet capillary. The vacuum seal to the MS ion guide draws all of the sample ions into the MS increasing the efficiency of sample transfer from the spray tip into the mass spectrometer. The direct connection to the inlet capillary eliminates the need for any source adjustment making the CaptiveSpray source truly Plug-and-Play.

CaptiveSpray Illustration

CaptiveSpray Illustration

CaptiveSpray Illustration

Structure elucidation

Structure elucidation

Structure elucidation

Tissue Proteomics for the Next Decade? Towards a Molecular Dimension in Histology

R Longuespee, M Fleron, C Pottier, F Quesada-Calvo, Marie-Alice Meuwis, et al.
OMICS A Journal of Integrative Biology 2014; 18(9)

Currently, sampling methods, biochemical procedures, and MS instrumentations allow scientists to perform ‘‘in depth’’ analysis of the protein content of any type of tissue of interest. This article reviews the salient issues in proteomics analysis of tissues. We first outline technical and analytical considerations for sampling and biochemical processing of tissues and subsequently the instrumental possibilities for proteomics analysis such as shotgun proteomics in an anatomical context. Specific attention concerns formalin fixed and paraffin embedded (FFPE) tissues that are potential ‘‘gold mines’’ for histopathological investigations. In all, the matrix assisted laser desorption/ionization (MALDI) MS imaging, which allows for differential mapping of hundreds of compounds on a tissue section, is currently the most striking evidence of linkage and transition between ‘‘classical’’ and ‘‘molecular’’ histology. Tissue proteomics represents a veritable field of research and investment activity for modern biomarker discovery and development for the next decade.

A transcriptome-proteome integrated network identifies ERp57 as a hub that mediates bone metastasis

N Santana-Codina, R Carretero, R Sanz-Pamplona1, T Cabrera, et al.
The American Society for Biochemistry and Molecular Biology
MCP  Apr 26, 2013; Manuscript M112.022772
E-mail: asierra@idibell.cat

Bone metastasis is the most common distant relapse in breast cancer. The identification of key proteins involved in the osteotropic phenotype would represent a major step toward the development of new prognostic markers and therapeutic improvements. The aim of this study was to characterize functional phenotypes that favor bone metastasis in human breast cancer.
We used the human breast cancer cell line MDA-MB-231 and its osteotropic BO2 subclone to identify crucial proteins in bone metastatic growth. We identified 31 proteins, 15 underexpressed and 16 overexpressed, in BO2 cells compared to parental cells. We employed a network-modeling approach in which these 31 candidate proteins were prioritized with respect to their potential in metastasis formation, based on the topology of the protein–protein interaction network and differential expression. The protein–protein interaction network provided a framework to study the functional relationships between biological molecules by attributing functions to genes whose functions had not been characterized.
The combination of expression profiles and protein interactions revealed an endoplasmic reticulum-thiol oxidoreductase, ERp57, functioning as a hub which retained 4 downregulated nodes involved in antigen presentation associated with the human major histocompatibility complex class I molecules, including HLA-A, HLA-B, HLA-E and HLA-F. Further analysis of the interaction network revealed an inverse correlation between ERp57 and vimentin, which influences cytoskeleton reorganization. Moreover, knockdown of ERp57 in BO2 cells confirmed its bone organ-specific prometastatic role. Altogether, ERp57 appears as a multifunctional chaperone that can regulate diverse biological processes to maintain the homeostasis of breast cancer cells and promote the development of bone metastasis.

Tandem-repeat protein domains across the tree of life

Kristin K. Jernigan and Seth R. Bordenstein
PeerJ 3:e732; 2015 http://dx.doi.org:/10.7717/peerj.732

Tandem-repeat protein domains, composed of repeated units of conserved stretches of 20–40 amino acids, are required for a wide array of biological functions. Despite their diverse and fundamental functions, there has been no comprehensive assessment of their taxonomic distribution, incidence, and associations with organismal lifestyle and phylogeny.
In this study, we assess for the first time the abundance of armadillo (ARM) and tetratricopeptide (TPR) repeat domains across all three domains in the tree of life and compare the results to our previous analysis on ankyrin (ANK) repeat domains in this journal. All eukaryotes and a majority of the bacterial and archaeal genomes analyzed have a minimum of one TPR and ARM repeat. In eukaryotes, the fraction of ARM-containing proteins is approximately double that of TPR and ANK-containing proteins, whereas bacteria and archaea are enriched in TPR-containing proteins relative to ARM- and ANK-containing proteins.
We show in bacteria that phylogenetic history, rather than lifestyle or pathogenicity, is a predictor of TPR repeat domain abundance, while neither phylogenetic history nor lifestyle predicts ARM repeat domain abundance. Surprisingly, pathogenic bacteria were not enriched in TPR-containing proteins, which have been associated within virulence factors in certain species. Taken together, this comparative analysis provides a newly appreciated view of the prevalence and diversity of multiple types of tandem-repeat protein domains across the tree of life.
A central finding of this analysis is that tandem repeat domain-containing proteins are prevalent not just in eukaryotes, but also in bacterial and archaeal species.

Detection of colorectal adenoma and cancer based on transthyretin and C3a-desArg serum levels

Anne-Kristin Fentz, Monika Sporl, Jorg Spangenberg, Heinz Joachim List, et al.
Proteomics Clin. Appl. 2007, 1, 536–544

Colorectal cancer is the second leading cause of cancer death, and it develops from benign colorectal adenomas in over 95% of patients. Early detection of these cancer precursors by screening tests and their removal can potentially eradicate more than 95% of colorectal cancers before they develop.
To discover sensitive and specific biomarkers for improvement of pre-clinical diagnosis of colorectal adenoma and cancer, we analysed in two independent studies (n = 87 and n = 83 patients) serum samples from colorectal cancer (stage III), colorectal adenoma and control patients using SELDI-TOF-MS. Extensive statistical analysis was performed to establish homogeneous patient groups based on their clinical data.
Two biomarkers that were each able to distinguish control patients from either colorectal adenoma or colorectal cancer patients (p,0.001) were identified as transthyretin (pre-albumin) and C3adesArg by MS/MS and were further validated by antibody-based assays (radial immunodiffusion, ELISA). A combination of both proteins clearly indicated the presence of colorectal adenoma or carcinoma. Using a cut-off of  >0.225 g/L for transthyretin and >1974 ng/mL for C3a-desArg, we found a sensitivity and specificity for colorectal adenoma of 96% and 70%, respectively.

The essential biology of the endoplasmic reticulum stress response for structural and computational biologists

Sadao Wakabayashi, Hiderou Yoshida
CSBJ Mar 2013; 6(7), e201303010   http://dx.doi.org/10.5936/csbj.201303010

The endoplasmic reticulum (ER) stress response is a cytoprotective mechanism that maintains homeostasis of the ER by upregulating the capacity of the ER in accordance with cellular demands. If the ER stress response cannot function correctly, because of reasons such as aging, genetic mutation or environmental stress, unfolded proteins accumulate in the ER and cause ER stress-induced apoptosis, resulting in the onset of folding diseases, including Alzheimer’s disease and diabetes mellitus. Although the mechanism of the ER stress response has been analyzed extensively by biochemists, cell biologists and molecular biologists, many aspects remain to be elucidated. For example, it is unclear how sensor molecules detect ER stress, or how cells choose the two opposite cell fates (survival or apoptosis) during the ER stress response. To resolve these critical issues, structural and computational approaches will be indispensable, although the mechanism of the ER stress response is complicated and difficult to understand holistically at a glance. Here, we provide a concise introduction to the mammalian ER stress response for structural and computational biologists.

Sequence co-evolution gives 3D contacts and structures of protein complexes

Thomas A Hopf, Charlotta P I Schärfe, João P G L M Rodrigues, et al.
eLife 2014;3:e03430   http://dx.doi.org:/10.7554/eLife.03430

Protein–protein interactions are fundamental to many biological processes. Experimental screens have identified tens of thousands of interactions, and structural biology has provided detailed functional insight for select 3D protein complexes. An alternative rich source of information about protein interactions is the evolutionary sequence record. Building on earlier work, we show that analysis of correlated evolutionary sequence changes across proteins identifies residues that are close in space with sufficient accuracy to determine the three-dimensional structure of the protein complexes. We evaluate prediction performance in blinded tests on 76 complexes of known 3D structure, predict protein–protein contacts in 32 complexes of unknown structure, and demonstrate how evolutionary couplings can be used to distinguish between interacting and non-interacting protein pairs in a large complex. With the current growth of sequences, we expect that the method can be generalized to genome-wide elucidation of protein–protein interaction networks and used for interaction predictions at residue resolution.
S-Glutathionylation of Cryptic Cysteines Enhances Titin Elasticity by Blocking Protein Folding

Jorge Alegre-Cebollada, P Kosuri, D Giganti, E Eckels, JA Rivas-Pardo, et al.
Cell, Mar 13, 2014; 156: 1235–1246. http://dx.doi.org/10.1016/j.cell.2014.01.056

The giant elastic protein titin is a determinant factor in how much blood fills the left ventricle during diastole and thus in the etiology of heart disease. Titin has been identified as a target of S-glutathionylation, an end product of the nitric-oxide-signaling cascade that increases cardiac muscle elasticity. However, it is unknown how S-glutathionylation may regulate the elasticity of titin and cardiac tissue.
Here, we show that mechanical unfolding of titin immunoglobulin (Ig) domains exposes buried cysteine residues, which then can be S-glutathionylated. S-glutathionylation of cryptic cysteines greatly decreases the mechanical stability of the parent Ig domain as well as its ability to fold. Both effects favor a more extensible state of titin. Furthermore, we demonstrate that S-glutathionylation of cryptic cysteines in titin mediates mechanochemical modulation of the elasticity of human cardiomyocytes.
We propose that posttranslational modification of cryptic residues is a general mechanism to regulate tissue elasticity.
Encounter complexes and dimensionality reduction in protein–protein association

Dima Kozakov, Keyong Li, David R Hall, Dmitri Beglov, Jiefu Zheng, et al.
eLife 2014;3:e01370 http://dx.doi.org:/10.7554/eLife.01370.001

An outstanding challenge has been to understand the mechanism whereby proteins associate. We report here the results of exhaustively sampling the conformational space in protein–protein association using a physics-based energy function. The agreement between experimental intermolecular paramagnetic relaxation enhancement (PRE) data and the PRE profiles calculated from the docked structures shows that the method captures both specific and non-specific encounter complexes. To explore the energy landscape in the vicinity of the native structure, the nonlinear manifold describing the relative orientation of two solid bodies is projected onto a Euclidean space in which the shape of low energy regions is studied by principal component analysis. Results show that the energy surface is canyon-like, with a smooth funnel within a two dimensional subspace capturing over 75% of the total motion. Thus, proteins tend to associate along preferred pathways, similar to sliding of a protein along DNA in the process of protein-DNA recognition.

Cardiovascular Proteomics: Evolution and Potential

  1. Kent Arrell, Irina Neverova and Jennifer E. Van Eyk
    Circ Res. 2001;88:763-773 http://dx.doi.org:/doi:/10.1161/hh0801.090193

The development of proteomics is a timely one for cardiovascular research. Analyses at the organ, subcellular, and molecular levels have revealed dynamic, complex, and subtle intracellular processes associated with heart and vascular disease. The power and flexibility of proteomic analyses, which facilitate protein separation, identification, and characterization, should hasten our understanding of these processes at the protein level. Properly applied, proteomics provides researchers with cellular protein “inventories” at specific moments in time, making it ideal for documenting protein modification due to a particular disease, condition, or treatment. This is accomplished through the establishment of species- and tissue-specific protein databases, providing a foundation for subsequent proteomic studies. Evolution of proteomic techniques has permitted more thorough investigation into molecular mechanisms underlying cardiovascular disease, facilitating identification not only of modified proteins but also of the nature of their modification. Continued development should lead to functional proteomic studies, in which identification of protein modification, in conjunction with functional data from established biochemical and physiological methods, has the ability to further our understanding of the interplay between proteome change and cardiovascular disease.

Advances in Proteomic Technologies and Its Contribution to the Field of Cancer

Mehdi Mesri

Advances in Medicine  2014, Article ID 238045, 25 pages http://dx.doi.org/10.1155/2014/238045

Systematic studies of the cancer genome have generated a wealth of knowledge in recent years. These studies have uncovered a number of new cancer genes not previously known to be causal targets in cancer. Genetic markers can be used to determine predisposition to tumor development, but molecularly targeted treatment strategies are not widely available for most cancers. Precision care plans still must be developed by understanding and implementing basic science research into clinical treatment. Proteomics is continuing to make major strides in the discovery of fundamental biological processes as well as more recent transition into an assay platform capable of measuring hundreds of proteins in any biological system. As such, proteomics can translate basic science discoveries into the clinical practice of precision medicine. The proteomic field has progressed at a fast rate over the past five years in technology, breadth and depth of applications in all areas of the bioscience. Some of the previously experimental technical approaches are considered the gold standard today, and the community is now trying to come to terms with the volume and complexity of the data generated. Here I describe contribution of proteomics in general and biological mass spectrometry in particular to cancer research, as well as related major technical and conceptual developments in the field.

Chemoproteomics reveals Toll-like receptor fatty acylation

Nicholas M Chesarino, Jocelyn C Hach, James L Chen, Balyn W Zaro, et al.
BMC Biology 2014, 12:91 http://www.biomedcentral.com/1741-7007/12/91

Background: Palmitoylation is a 16-carbon lipid post-translational modification that increases protein hydrophobicity. This form of protein fatty acylation is emerging as a critical regulatory modification for multiple aspects of cellular interactions and signaling. Despite recent advances in the development of chemical tools for the rapid identification and visualization of palmitoylated proteins, the palmitoyl proteome has not been fully defined. Here we sought to identify and compare the palmitoylated proteins in murine fibroblasts and dendritic cells.
Results: A total of 563 putative palmitoylation substrates were identified, more than 200 of which have not been previously suggested to be palmitoylated in past proteomic studies. Here we validate the palmitoylation of several new proteins including Toll-like receptors (TLRs) 2, 5 and 10, CD80, CD86, and NEDD4. Palmitoylation of TLR2, which was uniquely identified in dendritic cells, was mapped to a transmembrane domain-proximal cysteine. Inhibition of TLR2 S-palmitoylation pharmacologically or by cysteine mutagenesis led to decreased cell surface expression and a decreased inflammatory response to microbial ligands. Conclusions: This work identifies many fatty acylated proteins involved in fundamental cellular processes as well as cell type-specific functions, highlighting the value of examining the palmitoyl proteomes of multiple cell types. Spalmitoylation of TLR2 is a previously unknown immunoregulatory mechanism that represents an entirely novel avenue for modulation of TLR2 inflammatory activity.

Comparative Proteomics and Network Analysis Identify PKC Epsilon Underlying Long-Chain Fatty Acid Signaling

T Yonezawa, R Kurata, A Tajima, X Cui, H Maruta, H Nakaoka, K Nakajima and H Inokio
J Proteomics Bioinform 2014: 7:11 http://dx.doi.org/10.4172/jpb.1000337

Long-chain fatty acid possesses myriad roles in the biological function of the cells, not only as an energy substrate but also as substrates for cell membrane synthesis and as precursors for intracellular signaling molecules. However, little is known about the biological pathways that are stimulated by long-chain fatty acid. In order to identify the pathway of long-chain fatty acid, we performed 2-dimensional gel electrophoresis in the cells treated with or without oleate, and then analyzed 648 protein spots using PDQuest software and narrowed down 22 significant changing spots by statistical criterion. We also tried to determine these spots by MALDI-QIT-TOF-MS and SWISSPROT database query. We identified 11 proteins and predicted the biological network using available data sets from protein-protein interaction database. This prediction indicated that several protein kinase Cs (PKCs) underlie long chain fatty acid signaling. Indeed, oleate stimulated predicted PKC pathways. In expression array, oleate significantly up-regulated only PKC epsilon, but not other PKCs, in transcriptional levels. Collectively, our proteomics and network analysis implicates that PKC epsilon pathway plays an important role in long-chain fatty acid signaling.
Editorial: The art of proteomics translation

Translational Proteomics 2013; 1: 1–2 http://dx.doi.org/10.1016/j.trprot.2013.03.001

Over the years, the difficulties of transferring fundamental proteomics discoveries to clinical applications have caused a lot of frustration to proteomics researchers and clinicians alike, in both academia and industry. One of the reasons for this barrier is the lack of understanding between basic scientists and physicians: they have been trained using opposing concepts. Whilst the former want to control and understand all variables, the latter need rapid actions on patients, rather than absolute certainties. Both disciplines are difficult to con-dense into a single scientist and therefore interdisciplinary associations need to be fostered. Translational research has often been viewed as a two-way street: bedside to bench, and back to bedside. We should perhaps look at it as a roundabout, with the patient and his disease in the center, surrounded by a constant, iterative inter-play between basic, translational and clinical scientists, from both the public and private sectors. Proteomics research needs more than just a translation road bridge from discoveries to cures. Rather, it requires networks of road junctions to fill all the gaps and to allow cross-fertilization and synergies. Translational research and translational proteomics are more than just interesting concepts and hot keywords, they are supposed to improve the quality of people’s lives. With the launch of Translational Proteomics, we want to help the scientific and medical communities overcome the challenges on the long path from discovery to patient care. By focusing on connecting basic proteomics research to its ultimate clinical applications, the Journal will provide a space for publications detailing proteomics experiments, from early discovery to validation and the bedside.

Structural Basis of Diverse Membrane Target Recognitions by Ankyrins

C Wang, Z Wei, K Chen, F Ye, C Yu, V Bennett, and M Zhang
eLife 2014;  http:dx.doi.org:/10.7554/eLife.04353

Ankyrin adaptors together with their spectrin partners coordinate diverse ion channels and cell adhesion molecules within plasma membrane domains and  thereby promote physiological activities including fast signaling in the heart and  nervous system. Ankyrins specifically bind to numerous membrane targets through  their 24 ankyrin repeats (ANK repeats), although the mechanism for the facile and  independent evolution of these interactions has not been resolved. Here we report the structures of ANK repeats in complex with an inhibitory segment from the C-terminal regulatory domain and with a sodium channel Nav1.2 peptide, respectively, showing that the extended, extremely conserved inner groove spanning the entire ANK repeat solenoid contains multiple target binding sites capable of accommodating target protein with very diverse sequences via combinatorial usage of these sites. These structures establish a framework for understanding the evolution of ankyrins’ membrane targets, with implications for other proteins containing extended ANK repeat domains.

Fusion of Protein Aggregates Facilitates Asymmetric Damage Segregation

Miguel Coelho, Steven J. Lade, Simon Alberti, Thilo Gross, Iva M. Tolic
PLOS Biology June 2014; 12(6):e1001886

Asymmetric segregation of damaged proteins at cell division generates a cell that retains damage and a clean cell that supports population survival. In cells that divide asymmetrically, such as Saccharomyces cerevisiae, segregation of damaged proteins is achieved by retention and active transport. We have previously shown that in the symmetrically dividing Schizosaccharomyces pombe there is a transition between symmetric and asymmetric segregation of damaged proteins. Yet how this transition and generation of damage-free cells are achieved remained unknown. Here, by combining in vivo imaging of Hsp104-associated aggregates, a form of damage, with mathematical modeling, we find that fusion of protein aggregates facilitates asymmetric segregation. Our model predicts that, after stress, the increased number of aggregates fuse into a single large unit, which is inherited asymmetrically by one daughter cell, whereas the other one is born clean. We experimentally confirmed that fusion increases segregation asymmetry, for a range of stresses, and identified Hsp16 as a fusion factor. Our work shows that fusion of protein aggregates promotes the formation of damage-free cells. Fusion of cellular factors may represent a general mechanism for their asymmetric segregation at division.

Symmetric exchange of multi-protein building blocks between stationary focal adhesions and the cytosol

Jan-Erik Hoffmann, Y Fermin, R LO Stricker, K Ickstadt, E Zamir
eLife 2014;3:e02257. http://dx.doi.org:/10.7554/eLife.02257.001

How can the integrin adhesome get self-assembled locally, rapidly, and correctly as diverse cell-matrix adhesion sites? Here, we investigate this question by exploring the cytosolic state of integrin-adhesome components and their dynamic exchange between adhesion sites and cytosol. Using fluorescence cross-correlation spectroscopy (FCCS) and fluorescence recovery after photo-bleaching (FRAP) we found that the integrin adhesome is extensively pre-assembled already in the cytosol as multi-protein building blocks for adhesion sites. Stationary focal adhesions release symmetrically the same types of protein complexes that they recruit, thereby keeping the cytosolic pool of building blocks spatiotemporally uniform. We conclude a model in which multi-protein building blocks enable rapid and modular self-assembly of adhesion sites and symmetric exchange of these building blocks preserves their specifications and thus the assembly logic of the system.

Redox signaling via the molecular chaperone BiP protects cells against endoplasmic reticulum-derived oxidative stress

Jie Wang, Kristeen A Pareja, Chris A Kaiser, Carolyn S Sevier
eLife 2014;3:e03496. http://dx.doi.org:/10.7554/eLife.03496

Oxidative protein folding in the endoplasmic reticulum (ER) has emerged as a potentially significant source of cellular reactive oxygen species (ROS). Recent studies suggest that levels of ROS generated as a byproduct of oxidative folding rival those produced by mitochondrial respiration. Mechanisms that protect cells against oxidant accumulation within the ER have begun to be elucidated yet many questions still remain regarding how cells prevent oxidant-induced damage from ER folding events. Here we report a new role for a central well-characterized player in ER homeostasis as a direct sensor of ER redox imbalance. Specifically we show that a conserved cysteine in the lumenal chaperone BiP is susceptible to oxidation by peroxide, and we demonstrate that oxidation of this conserved cysteine disrupts BiP’s ATPase cycle. We propose that alteration of BiP activity upon oxidation helps cells cope with disruption to oxidative folding within the ER during oxidative stress.

Current perspectives on cadherin-cytoskeleton interactions and dynamics

Xuan Liang, Guillermo A Gomez, Alpha S Yap
Cell Health and Cytoskeleton 2015:7 11–24

Cells are linked together dynamically by adhesion molecules, such as the classical cadherins. E-cadherin, which mediates epithelial cell–cell interactions, plays fundamental roles in tissue organization and is often perturbed in diseases such as cancer. It has long been recognized that the biology of E-cadherin arises from cooperation between adhesion and the actin cytoskeleton. A major feature is the generation of contractile forces at junctions, yielding patterns of tension that contribute to tissue integrity and patterning. Here we discuss recent developments in understanding how cadherin junctions integrate signaling and cytoskeletal dynamics to sense and generate force.

N-glycosylation status of E-cadherin controls cytoskeletal dynamics through the organization of distinct β-catenin- and γ-catenin-containing AJs

Basem T Jamal, M Nita-Lazar, Z Gao, B Amin, J Walker, MA Kukuruzinska
Cell Health and Cytoskeleton 2009:1 67–80

N-glycosylation of E-cadherin has been shown to inhibit cell–cell adhesion. Specifically, our recent studies have provided evidence that the reduction of E-cadherin N-glycosylation promoted the recruitment of stabilizing components, vinculin and serine/threonine protein phosphatase 2A (PP2A), to adherens junctions (AJs) and enhanced the association of AJs with the actin cytoskeleton. Here, we examined the details of how N-glycosylation of E-cadherin affected the molecular organization of AJs and their cytoskeletal interactions. Using the hypoglycosylated E-cadherin variant, V13, we show that V13/β-catenin complexes preferentially interacted with PP2A and with the microtubule motor protein dynein. This correlated with dephosphorylation of the microtubule-associated protein tau, suggesting that increased association of PP2A with V13-containing AJs promoted their tethering to microtubules. On the other hand, V13/γ-catenin complexes associated more with vinculin, suggesting that they mediated the interaction of AJs with the actin cytoskeleton. N-glycosylation driven changes in the molecular organization of AJs were physiologically significant because transfection of V13 into A253 cancer cells, lacking both mature AJs and tight junctions (TJs), promoted the formation of stable AJs and enhanced the function of TJs to a greater extent than wild-type E-cadherin. These studies provide the first mechanistic insights into how N-glycosylation of E-cadherin drives changes in AJ composition through the assembly of distinct β-catenin- and γ-catenin-containing scaffolds that impact the interaction with different cytoskeletal components.

Mapping the dynamics of force transduction at cell-cell 4 junctions of epithelial clusters

Mei Rosa Ng, Achim Besser, Joan S. Brugge, Gaudenz Danuser
eLife 2014;10.7554/eLife.03282

Force transduction at cell-cell adhesions regulates tissue development, maintenance and adaptation. We developed computational and experimental approaches to quantify, with both subcellular and multi-cellular resolution, the dynamics of force transmission in cell clusters. Applying this technology to spontaneously-forming adherent epithelial cell clusters, we found that basal force fluctuations were coupled to E-cadherin localization at the level of individual cell-cell junctions. At the multi-cellular scale, cell-cell force exchange depended on the cell position within a cluster, and was adaptive to reconfigurations due to cell divisions or positional rearrangements. Importantly, force transmission through a cell required coordinated modulation of cell-matrix adhesion and actomyosin contractility in the cell and its neighbors. These data provide insights into  mechanisms that could control mechanical stress homeostasis in dynamic epithelial tissues, and highlight our methods as a resource for the study of mechanotransduction in cell-cell adhesions.

G-protein-coupled receptor signaling and polarized actin dynamics drive cell-in-cell invasion

Vladimir Purvanov, Manuel Holst, Jameel Khan, Christian Baarlink, Robert Grosse
eLife 2014;3:e02786.  http://dx.doi.org:/10.7554/eLife.02786

Homotypic or entotic cell-in-cell invasion is an integrin-independent process observed in carcinoma cells exposed during conditions of low adhesion such as in exudates of malignant disease. Although active cell-in-cell invasion depends on RhoA and actin, the precise mechanism as well as the underlying actin structures and assembly factors driving the process are unknown. Furthermore, whether specific cell surface receptors trigger entotic invasion in a signal-dependent fashion has not been investigated. In this study, we identify the G-protein-coupled LPA receptor 2 (LPAR2) as a signal transducer specifically required for the actively invading cell during entosis. We find that G12/13 and PDZ-RhoGEF are required for entotic invasion, which is driven by blebbing and a uropod-like actin structure at the rear of the invading cell. Finally, we provide evidence for an involvement of the RhoA-regulated formin Dia1 for entosis downstream of LPAR2. Thus, we delineate a signaling process that regulates actin dynamics during cell-in-cell invasion.

Cytoskeletal Basis of Ion Channel Function in Cardiac Muscle

Matteo Vatta, and Georgine Faulkner
Future Cardiol. 2006 Jul 1; 2(4): 467–476. http://dx.doi.org:/10.2217/14796678.2.4.467

The heart is a force-generating organ that responds to self-generated electrical stimuli from specialized cardiomyocytes. This function is modulated by sympathetic and parasympathetic activity.

In order to contract and accommodate the repetitive morphological changes induced by the cardiac cycle, cardiomyocytes depend on their highly evolved and specialized cytoskeletal apparatus. Defects in components of the cytoskeleton, in the long term, affect the ability of the cell to compensate at both functional and structural levels. In addition to the structural remodeling, the myocardium becomes increasingly susceptible to altered electrical activity leading to arrhythmogenesis. The development of arrhythmias secondary to structural remodeling defects has been noted, although the detailed molecular mechanisms are still elusive. Here I will review the current knowledge of the molecular and functional relationships between the cytoskeleton and ion channels and, I will discuss the future impact of new data on molecular cardiology research and clinical practice.

Structure and transport mechanism of the sodium/proton 2 antiporter MjNhaP1

Cristina Paulino, D Wöhlert , E Kapotova, Ö Yildiz & W Kühlbrandt
eLife 2014;  http://dx.doi.org/10.7554/eLife.03583

Sodium/proton antiporters are essential for sodium and pH homeostasis and play a major role in human health and disease. We determined the structures of the archaeal sodium/proton antiporter MjNhaP1 in two complementary states. The inward-open state was obtained by x-ray crystallography in the presence of sodium at pH8, where the transporter is highly active. The outward-open state was obtained by electron crystallography without sodium at pH4, where MjNhaP1 is inactive. Comparison of both structures reveals a 7° tilt of the 6-helix bundle. Na+  uptake measurements indicate non-cooperative transport with an activity maximum at pH7.5. We conclude that binding of a Na+ ion from the outside induces helix movements that close the extracellular cavity, open the cytoplasmic funnel, and result in a ~5 Å vertical relocation of the ion binding site to release the substrate ion into the cytoplasm.

Integrated control of transporter endocytosis and recycling by the arrestin-related protein Rod1 and the ubiquitin ligase Rsp5

Michel Becuwe, Sébastien Léon
eLife 2014; http://dx.doi.org/10.7554/eLife.03307

After endocytosis, membrane proteins can recycle to the cell membrane or be degraded in lysosomes. Cargo ubiquitylation favors their lysosomal targeting and can be regulated by external signals, but the mechanism is ill-defined. Here, we studied the post-endocytic trafficking of Jen1, a yeast monocarboxylate transporter, using microfluidics-assisted live cell imaging. We show that the ubiquitin ligase Rsp5 and the glucose-regulated arrestin related (ART) protein Rod1, involved in the glucose-induced internalization of Jen1, are  also required for the post-endocytic sorting of Jen1 to the yeast lysosome. This new step takes place at the trans-Golgi network (TGN), where Rod1 localizes dynamically upon triggering endocytosis. Indeed, transporter trafficking to the TGN after internalization is required for their degradation. Glucose removal promotes Rod1 relocalization to the cytosol and Jen1 deubiquitylation, allowing transporter recycling when the signal is only transient. Therefore, nutrient availability regulates transporter fate through the localization of the ART/Rsp5 ubiquitylation complex at the TGN.

  1. McKenney, W Huynh, ME. Tanenbaum, G Bhabha, and RD. Vale
    Science Express 19 June 2014 /10.1126/science.1254198

Cytoplasmic dynein is a molecular motor that transports a large variety of cargoes (e.g., organelles, mRNAs, and viruses) along microtubules over long intracellular distances. The dynactin protein complex is important for dynein activity in vivo, but its precise role has been unclear. Here, we found that purified mammalian dynein did not move processively on microtubules in vitro. However, when dynein formed a complex with dynactin and one of four different cargo-specific adapter proteins, the motor became ultra-processive, moving for distances similar to those of native cargoes in living cells. Thus, we propose that dynein is largely inactive in the cytoplasm and that a variety of adapter proteins activate processive motility by linking dynactin to dynein only when the motor is bound to its proper cargo.

Removal of surface charge–charge interactions from ubiquitin leaves the protein folded and very stable

Vakhtang V. Loladze And George I. Makhatadze
Protein Science (2002), 11:174–177

The contribution of solvent-exposed charged residues to protein stability was evaluated using ubiquitin as a model protein. We combined site-directed mutagenesis and specific chemical modifications to first replace all Arg residues with Lys, followed by carbomylation of Lys- amino groups. Under the conditions in which all carboxylic groups are protonated (at pH 2), the chemically modified protein is folded and very stable (dG= 18 kJ/mol). These results indicate that surface charge–charge interactions are not an essential fundamental force for protein folding and stability.

Phase Transitions of Multivalent Proteins Can Promote Clustering of Membrane Receptors

Sudeep Banjade and Michael K. Rosen
eLife 2014; http://dx.doi.org/10.7554/eLife.04123

Clustering of proteins into micrometer-sized structures at membranes is observed in many signaling pathways. Most models of clustering are specific to particular systems, and relationships between physical properties of the clusters and their molecular components are not well understood. We report biochemical reconstitution on supported lipid bilayers of protein clusters containing the adhesion receptor Nephrin, and its cytoplasmic partners, Nck and N-WASP. With Nephrin attached to the bilayer, multivalent interactions enable these proteins to polymerize on the membrane surface and undergo two-dimensional phase separation, producing micrometer-sized clusters. Dynamics and thermodynamics of the clusters are modulated by the valencies and affinities of the interacting species. In the presence of the Arp2/3 complex, the clusters assemble actin filaments, suggesting that clustering of regulatory factors could promote local actin assembly at membranes. Interactions between multivalent proteins could be a  general mechanism for cytoplasmic adaptor proteins to organize membrane receptors into micrometer-scale signaling zones.

The quantitative architecture of centromeric chromatin

Dani L Bodor, João F Mata, Mikhail Sergeev, Ana Filipa David, et al.
eLife 2014;3:e02137. http://dx.doi.org:/10.7554/eLife.02137

The centromere, responsible for chromosome segregation during mitosis, is epigenetically defined by CENP-A containing chromatin. The amount of centromeric CENP-A has direct implications for both the architecture and epigenetic inheritance of centromeres. Using complementary strategies, we determined that typical human centromeres contain ∼400 molecules of CENP-A, which is controlled by a mass-action mechanism. This number, despite representing only ∼4% of all centromeric nucleosomes, forms a ∼50-fold enrichment to the overall genome. In addition, although pre-assembled CENP-A is randomly segregated during cell division, this amount of CENP-A is sufficient to prevent stochastic loss of centromere function and identity. Finally, we produced a statistical map of CENP-A occupancy at a human neocentromere and identified nucleosome positions that feature CENP-A in a majority of cells. In summary, we present a quantitative view of the centromere that provides a mechanistic framework for both robust epigenetic inheritance of centromeres and the paucity of neocentromere formation.

Synaptic proteins promote calcium-triggered fast transition from point contact to full fusion

Jiajie Diao, Patricia Grob, Daniel J Cipriano, Minjoung Kyoung
eLife 2012;1:e00109. http://dx.doi.org:/10.7554/eLife.00109

The molecular underpinnings of synaptic vesicle fusion for fast neurotransmitter release are still unclear. Here, we used a single vesicle–vesicle system with reconstituted SNARE and synaptotagmin-1 proteoliposomes to decipher the temporal sequence of membrane states upon Ca2+-injection at 250–500 μM on a 100-ms timescale. Furthermore, detailed membrane morphologies were imaged with cryo-electron microscopy before and after Ca2+-injection. We discovered a heterogeneous network of immediate and delayed fusion pathways. Remarkably, all instances of Ca2+-triggered immediate fusion started from a membrane–membrane point-contact and proceeded to complete fusion without discernible hemifusion intermediates. In contrast, pathways that involved a stable hemifusion diaphragm only resulted in fusion after many seconds, if at all. When complexin was included, the Ca2+-triggered fusion network shifted towards the immediate pathway, effectively synchronizing fusion, especially at lower Ca2+-concentration. Synaptic proteins may have evolved to select this immediate pathway out of a heterogeneous network of possible membrane fusion pathways.

Cytoskeleton, cytoskeletal interactions, and vascular endothelial function

Jingli Wang, Michael E Widlansky
Cell Health and Cytoskeleton 2012:4 119–127

Far from being inert, the vascular endothelium is a critical regulator of vascular function. While the endothelium participates in autocrine, paracrine, and endocrine signaling, it also transduces mechanical signals from the cell surface involving key cell structural elements. In this review, we discuss the structure of the vascular endothelium and its relationship to traditional cardiovascular risk factors and clinical cardiovascular events. Further, we review the emerging evidence that cell structural elements, including the glycocalyx, intercellular junctions, and cytoskeleton elements, help the endothelium to communicate with its environment to regulate vascular function, including vessel permeability and signal transduction via nitric oxide bioavailability. Further work is necessary to better delineate the regulatory relationships between known key regulators of vascular function and endothelial cell structural elements.

Cellular prion protein is required for neuritogenesis: fine-tuning of multiple signaling pathways involved in focal adhesions and actin cytoskeleton dynamics

Aurélie Alleaume-Butaux, C Dakowski, M Pietri, S Mouillet-Richard, et al.
Cell Health and Cytoskeleton 2013:5 1–12

Neuritogenesis is a dynamic phenomenon associated with neuronal differentiation that allows a rather spherical neuronal stem cell to develop dendrites and axon, a prerequisite for the integration and transmission of signals. The acquisition of neuronal polarity occurs in three steps: (1) neurite sprouting, which consists of the formation of buds emerging from the postmitotic neuronal soma; (2) neurite outgrowth, which represents the conversion of buds into neurites, their elongation and evolution into axon or dendrites; and (3) the stability and plasticity of neuronal polarity. In neuronal stem cells, remodeling and activation of focal adhesions (FAs) associated with deep modifications of the actin cytoskeleton is a prerequisite for neurite sprouting and subsequent neurite outgrowth. A multiple set of growth factors and interactors located in the extracellular matrix and the plasma membrane orchestrate neuritogenesis by acting on intracellular signaling effectors, notably small G proteins such as RhoA, Rac, and Cdc42, which are involved in actin turnover and the dynamics of FAs. The cellular prion protein (PrPC), a glycosylphosphatidylinositol (GPI)-anchored membrane protein mainly known for its role in a group of fatal neurodegenerative diseases, has emerged as a central player in neuritogenesis. Here, we review the contribution of PrPC to neuronal polarization and detail the current knowledge on the signaling pathways fine-tuned by PrPC to promote neurite sprouting, outgrowth, and maintenance. We emphasize that PrPC-dependent neurite sprouting is a process in which PrPC governs the dynamics of FAs and the actin cytoskeleton via β1 integrin signaling. The presence of PrPC is necessary to render neuronal stem cells competent to respond to neuronal inducers and to develop neurites. In differentiating neurons, PrPC exerts a facilitator role towards neurite elongation. This function relies on the interaction of PrPC with a set of diverse partners such as elements of the extracellular matrix, plasma membrane receptors, adhesion molecules, and soluble factors that control actin cytoskeleton turnover through Rho-GTPase signaling. Once neurons have reached their terminal stage of differentiation and acquired their polarized morphology, PrPC also takes part in the maintenance of neurites. By acting on tissue nonspecific alkaline phosphatase, or matrix metalloproteinase type 9, PrPC stabilizes interactions between neurites and the extracellular matrix.

Broader implications: biological and clinical significance of microtubule acetylation

Sharon M Rymut, Thomas J Kelley
Cell Health and Cytoskeleton 2015:7 71–82

Microtubule acetylation is a key posttranslational modification that enhances organelle transport, drives cell signaling, and regulates cell cycle regulation. The optimal level of microtubule acetylation is regulated by the acetyltransferase alpha-tubulin-N-acetyltransferase 1and two deacetylases, histone deacetylase 6 and sirtuin-2. Alterations in microtubule acetylation levels have been associated with the pathophysiology of a number of diseases, including various forms of neurodegenerative conditions, cancer, and even cystic fibrosis. In this review, we will highlight the biological and clinical significance of microtubule acetylation and the potential of targeting this pathway for therapeutics.

Inositol-1,4,5-trisphosphate 1 (IP3)-mediated STIM1 oligomerization requires  intact mitochondrial Ca2+ uptake

  1. Deak, S. Blass, M. J. Khan, L. N. Groschner, M. Waldeck-Weiermair, et al.
    Journal of Cell Science 2014 advanced print

Mitochondria contribute to cell signaling by controlling store-operated Ca2+ entry (SOCE).  SOCE is activated by Ca2+ release from the endoplasmic reticulum (ER), whereupon the stromal  interacting molecule 1 (STIM1) forms oligomers, redistributes to ER-plasma membrane  junctions, and opens plasma membrane Ca2+ channels. Mechanisms by which mitochondria interfere with the complex process of SOCE are insufficiently clarified. In this study we used a shRNA approach to investigate the direct involvement of mitochondrial Ca2+ buffering in SOCE. We demonstrate that knock-down of two proteins that are essential for mitochondrial Ca2+ uptake, either the mitochondrial calcium uniporter (MCU) or uncoupling protein 2 (UCP2), results in decelerated STIM1 oligomerization and impaired SOCE following cell stimulation with an inositol-1,4,5-trisphosphate (IP3)-generating agonist. Upon artificially augmented cytosolic Ca2+-buffering or ER Ca2+ depletion by sarco/endoplasmic reticulum Ca2+-ATPase (SERCA) inhibitors, STIM1 oligomerization did not rely on intact mitochondrial Ca2+ uptake.  However, MCU-dependent mitochondrial sequestration of Ca2+ entering through the SOCE  pathway was essential to prevent slow deactivation of SOCE. Our findings show a stimulus specific contribution of mitochondrial Ca2+ uptake to the SOCE machinery likely by shaping cytosolic Ca2+ micro-domains.

Role of forkhead box protein A3 in age-associated metabolic decline

Xinran Ma, Lingyan Xu, Oksana Gavrilov, and Elisabetta Mueller
PNAS | September 30, 2014 | vol. 111 | no. 39 | 14289–14294

Aging is associated with increased adiposity and diminished thermogenesis, but the critical transcription factors influencing these metabolic changes late in life are poorly understood. We recently demonstrated that the winged helix factor forkhead box protein A3 (Foxa3) regulates the expansion of visceral adipose tissue in high-fat diet regimens; however, whether Foxa3 also contributes to the increase in adiposity and the decrease in brown fat activity observed during the normal aging process is currently unknown.
Here we report that during aging, levels of Foxa3 are significantlyand selectively up-regulated in brown and inguinal white fat depots, and that midage Foxa3-null mice have increased white fat browning and thermogenic capacity, decreased adipose tissue expansion, improved insulin sensitivity, and increased longevity. Foxa3 gain-of-function and loss-of-function studies in inguinal adipose depots demonstrated a cell-autonomous function for Foxa3 in white fat tissue browning. Furthermore, our analysis revealed that the mechanisms of Foxa3 modulation of brown fat gene programs involve the suppression of peroxisome proliferator activated receptor γ coactivtor 1 α (PGC1α) levels through interference with cAMP responsive element binding protein 1-mediated transcriptional regulation of the PGC1α promoter. Overall, our data demonstrate a role for Foxa3 in energy expenditure and in age-associated metabolic disorders.

Prediction of enzyme function by combining sequence similarity and protein interactions

Jordi Espadaler, Narayanan Eswa, Enrique Querol, Francesc X Avilés, et al.
BMC Bioinformatics 2008, 9:249 http://dx.doi.org:/10.1186/1471-2105-9-249

Background: A number of studies have used protein interaction data alone for protein function prediction. Here, we introduce a computational approach for annotation of enzymes, based on the observation that similar protein sequences are more likely to perform the same function if they share similar interacting partners.
Results: The method has been tested against the PSI-BLAST program using a set of 3,890 protein sequences from which interaction data was available. For protein sequences that align with at least 40% sequence identity to a known enzyme, the specificity of our method in predicting the first three EC digits increased from 80% to 90% at 80% coverage when compared to PSI-BLAST.
Conclusion: Our method can also be used in proteins for which homologous sequences with known interacting partners can be detected. Thus, our method could increase 10% the specificity of genome-wide enzyme predictions based on sequence matching by PSI-BLAST alone.

Plasma Transthyretin Indicates the Direction of both Nitrogen Balance and Retinoid Status in Health and Disease

Ingenbleek Yves and Bienvenu Jacques
The Open Clinical Chemistry Journal, 2008, 1, 1-12

Whatever the nutritional status and the disease condition, the actual transthyretin (TTR) plasma level is determined by opposing influences between anabolic and catabolic alterations. Rising TTR values indicate that synthetic processes prevail over tissue breakdown with a nitrogen balance (NB) turning positive as a result of efficient nutritional support and / or anti-inflammatory therapy. Declining TTR values point to the failure of sustaining NB as an effect of maladjusted dietetic management and / or further worsening of the morbid condition. Serial measurement of TTR thus appears as a dynamic index defining the direction of NB in acute and chronic disorders, serving as a guide to alert the physician on the validity of his therapeutic strategy. The level of TTR production by the liver also works as a limiting factor for the cellular bioavailability of retinol and retinoid derivatives which play major roles in the brain ageing process. Optimal protein nutritional status, as assessed by TTR values within the normal range, prevents the occurrence of vascular and cerebral damages while maintaining the retinoid-mediated memory, cognitive and behavioral activities of elderly persons.

Prof. Dr. Volker Haucke
Institut für Chemie-Biochemie
Takustrasse 6

Eukaryotic cells contain three major types of cytoskeletal filaments

Eukaryotic cells contain three major types of cytoskeletal filaments

major types of cytoskeletal filaments

major types of cytoskeletal filaments

Intermediate Filaments support the nuclear membrane and connect cells at cell junctions

Intermediate Filaments support the nuclear membrane and connect cells at cell junctions

microtubules (MTs; green) radiate from MTOCs (yellow) towards the cell periphery

microtubules (MTs; green) radiate from MTOCs (yellow) towards the cell periphery

Actin polymerization in vitro reveals a critical dependence of filament assembly on G-actin concentration via a 3-step nucleation mechanism

Actin polymerization in vitro reveals a critical dependence of filament assembly on G-actin concentration via a 3-step nucleation mechanism

Binding-proteins and receptors

Motor, visual and emotional deficits in mice after closed-head mild traumatic brain injury are alleviated by the novel CB2 inverse agonist SMM-189
Reiner, A., Heldt, S.A., Presley, C.S., (…), Gurley, S.N., Moore, B.M.
2015  International Journal of Molecular Sciences 16 (1), pp. 758-787

We have developed a focal blast model of closed-head mild traumatic brain injury (TBI) in mice. As true for individuals that have experienced mild TBI, mice subjected to 50-60 psi blast show motor, visual and emotional deficits, diffuse axonal injury and microglial activation, but no overt neuron
loss. Because microglial activation can worsen brain damage after a concussive event and because microglia can be
modulated by their cannabinoid type 2 receptors (CB2), we evaluated the effectiveness of the novel CB2 receptor inverse agonist SMM-189 in altering microglial activation and mitigating deficits after mild TBI. In vitro analysis indicated that SMM-189 converted human microglia from the pro-inflammatory M1 phenotype to the pro-healing M2 phenotype. Studies in mice showed that daily administration of SMM-189 for two weeks beginning shortly after blast greatly reduced the motor, visual, and emotional deficits otherwise evident after 50-60 psi blasts, and prevented brain injury that may contribute to these deficits. Our results suggest that treatment with the CB2 inverse agonist SMM-189 after a mild TBI event can reduce its adverse consequences by beneficially modulating microglial activation. These
findings recommend further evaluation of CB2 inverse agonists as a novel therapeutic approach for treating mild TBI.

The novel small leucine-rich protein chondroadherin-like (CHADL) is expressed in cartilage and modulates chondrocyte differentiation
Tillgren, V., Ho, J.C.S., Önnerfjord, P., Kalamajski, S.
2015  Journal of Biological Chemistry 290 (2), pp. 918-925

The constitution and biophysical properties of extracellular matrices can dramatically influence cellular phenotype during development, homeostasis, or pathogenesis. These effects can be signaled through a differentially regulated assembly of collagen fibrils, orchestrated by a family of collagen-associated small leucine-rich proteins (SLRPs). In this report, we describe the tissue-specific expression and function of a previously uncharacterized SLRP, chondroadherin-like (CHADL). We developed antibodies against CHADL and, by immunohistochemistry, detected CHADL expression mainly in skeletal tissues, particularly in fetal cartilage and in the pericellular space of adult chondrocytes. In situ hybridizations and immunoblots on tissue lysates confirmed this tissue-specific expression pattern. Recombinant CHADL bound collagen in cell culture and inhibited in vitro collagen fibrillogenesis. After Chadl shRNA knockdown, chondrogenic ATDC5 cells increased their differentiation, indicated by increased transcript levels of Sox9, Ihh, Col2a1, and Col10a1. The knockdown increased collagen II and aggrecan deposition in the cell layers.

Microarray analysis of the knockdown samples suggested collagen receptor-related changes, although other upstream effects could not be excluded. Together, our data indicate that the novel SLRP CHADL is expressed in cartilaginous tissues, influences collagen fibrillogenesis, and modulates chondrocyte differentiation. CHADL appears to have a negative regulatory role, possibly ensuring the formation of a stable extracellular matrix.

P53 protein-mediated Up-regulation of MAP kinase phosphatase 3 (MKP-3) contributes to the establishment of the cellular senescent phenotype through dephosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2)
Zhang, H., Chi, Y., Gao, K., Zhang, X., Yao, J.
2015  Source of the DocumentJournal of Biological Chemistry 290 (2), pp. 1129-1140

Growth arrest is one of the essential features of cellular senescence. At present, the precise mechanisms responsible for the establishment of the senescence-associated arrested phenotype are still incompletely understood. Given that ERK1/2 is one of the major kinases controlling cell growth and proliferation, we examined the possible implication of ERK1/2. Exposure of normal rat epithelial cells to etoposide caused cellular senescence, as manifested by enlarged cell size, a flattened cell body, reduced cell proliferation, enhanced ?-galactosidase activity, and elevated p53 and p21. Senescent cells displayed a blunted response to growth factor-induced cell proliferation, which was preceded by impaired ERK1/2 activation. Further analysis revealed that senescent cells expressed a significantly higher level of mitogenactivated protein phosphatase 3 (MKP-3, a cytosolic ERK1/2-targeted phosphatase), which was suppressed by blocking the transcriptional activity of the tumor suppressor p53 with pifithrin-?. Inhibition of MKP-3 activity with a specific inhibitor or siRNA enhanced basal ERK1/2 phosphorylation and promoted cell proliferation. Apart from its role in growth arrest, impairment of ERK1/2 also contributed to the resistance of senescent cells to oxidant-elicited cell injury. These results therefore indicate that p53-mediated up-regulation of MKP-3 contributes to the establishment of the senescent cellular phenotype through dephosphorylating ERK1/2. Impairment of ERK1/2 activation could be an important mechanism by which p53 controls cellular senescence.

Dynamics and interaction of Interleukin-4 receptor subunits in living cells
Gandhi, H., Worch, R., Kurgonaite, K., (…), Bökel, C., Weidemann, T.
2015  Biophysical Journal 107 (11), pp. 2515-2527

It has long been established that dimerization of Interleukin-4 receptor (IL-4R) subunits is a pivotal step for JAK/STAT signal transduction. However, ligand-induced complex formation at the surface of living cells has been challenging to observe. Here we report an experimental assay employing trisNTA dyes for orthogonal, external labeling of eGFP-tagged receptor constructs that allows the quantification of receptor heterodimerization by dual-color fluorescence cross-correlation spectroscopy. Fluorescence cross-correlation spectroscopy analysis at the plasma membrane shows that IL-4R subunit dimerization is indeed a strictly ligand-induced process.

Under conditions of saturating cytokine occupancy, we determined intramembrane dissociation constants (Kd,2D) of 180 and 480 receptors per ?m2 for the type-2 complexes IL-4:IL-4R?/IL-13R?1 and IL-13:IL-13R?1/IL-4R?, respectively. For the lower affinity type-1 complex IL-4:IL-4R?/IL-2R?, we estimated a Kd,2D of ?1000 receptors per ?m2. The receptor densities required for effective dimerization thus exceed the typical, average expression levels by several orders of magnitude. In addition, we find that all three receptor subunits accumulate rapidly within a subpopulation of early sorting and recycling endosomes stably anchored just beneath the plasma membrane (cortical endosomes, CEs). The receptors, as well as labeled IL-4 and trisNTA ligands are specifically trafficked into CEs by a constitutive internalization mechanism. This may compensate for the inherent weak affinities that govern ligand-induced receptor dimerization at the plasma membrane. Consistently, activated receptors are also concentrated at the CEs. Our observations thus suggest that receptor trafficking may play an important role for the regulation of IL-4R-mediated JAK/STAT signaling.

Role of mitochondria in nonalcoholic fatty liver disease
Nassir, F., Ibdah, J.A.
2015  International Journal of Molecular Sciences 15 (5), pp. 8713-8742

Nonalcoholic fatty liver disease (NAFLD) affects about 30% of the general population in the United States and includes a spectrum of disease that includes simple steatosis, non-alcoholic steatohepatitis (NASH), fibrosis and cirrhosis. Significant insight has been gained into our understanding of the pathogenesis of NALFD; however the key metabolic aberrations underlying lipid accumulation in hepatocytes and the progression of NAFLD remain to be elucidated. Accumulating and emerging evidence indicate that hepatic mitochondria play a critical role in the development and pathogenesis of steatosis and NAFLD. Here, we review studies that document a link between the pathogenesis of NAFLD and hepatic mitochondrial dysfunction with particular focus on new insights into the role of impaired fatty acid oxidation, the transcription factor peroxisome proliferator-activated receptor-? coactivator-1? (PGC-1?), and sirtuins in development and progression of NAFLD.

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Natural Products Chemistry

Writer and Curator: Larry H. Bernstein, MD, FCAP 



Natural products chemistry or pharmacognosy, the study of the physical, chemical, biochemical and biological properties of drugs, drug substances or potential drugs

or drug substances of natural origin as well as the search for new drugs from natural
sources, is an a tradition in medicine that reaches to a tradition thousands of years
old.  It has to some extent been supplanted by structural organic chemistry, metallo-organic chemistry, and synthetic organic chemistry of families of drugs.  In some
cases, drug failures may be attributed to the inherent failure in a family, and in others
there has been substitution of a drug compound by another with eaqual or greater
potency and less toxicity. A serious confounder has been that medications intended
for a specific effect has either an unfavorable interaction with another class of drugs,
or it has a metabolic reaction with another organ or pathway than the use intended.
That has been the huge impediment to pharmaceutical development.

However, it is important to remember that many of the medications in common use
were originally plant or natural derivatives, e.g., digoxin, Warfarin.

Thymoquinone, an extract of nigella sativa seed oil, blocked pancreatic cancer cell
growth and killed the cells by enhancing the process of programmed cell death
Steve Benowitz  steven.benowitz@jefferson.edu

Researchers at the Kimmel Cancer at Jefferson in Philadelphia have found that
thymoquinone, an extract of nigella sativa seed oil, blocked pancreatic cancer cell
growth and killed the cells by enhancing the process of programmed cell death.
According to Hwyda Arafat, M.D., Ph.D., associate professor of Surgery at
Jefferson Medical College of Thomas Jefferson University, nigella sativa helps treat
a broad array of diseases, including some immune and inflammatory disorders.
Previous studies also have shown anticancer activity in prostate and colon cancers,
as well as antioxidant and anti-inflammatory effects.

Using a human pancreatic cancer cell line, she and her team found that adding
thymoquinone killed approximately 80 percent of the cancer cells. They demonstrated
that thymoquinone triggered programmed cell death in the cells, and that a number of
important genes, including p53, Bax, bcl-2 and p21, were affected. The researchers
found that expression of p53, a tumor suppressor gene, and Bax, a gene that promotes
programmed cell death, was increased, while bcl-2, which blocks such cell death,
was decreased. The p21 gene, which is involved in the regulation of different phases
of the cell cycle, was substantially increased.

In addition, adding thymoquinone to pancreatic cancer cells reduced the production
and activity of enzymes called histone deacetylases (HDACs), which remove the
acetyl groups from the histone proteins, halting the gene transcription process.
Dr. Arafat notes that HDAC inhibitors are a “hot” new class of drugs that interfere
with the function of histone deacetylases, and is being studied as a treatment for
cancer and neurodegenerative diseases.

Extra Virgin Olive Oil Improves Learning and Memory in SAMP8 Mice
SA Farra, TO Price, LJ Dominguez, A Motisi, F Saianoe, et al.
Journal of Alzheimer’s Disease 28 (2012) 81–92

Polyphenols are potent antioxidants found in extra virgin olive oil (EVOO);
antioxidants have been shown to reverse age- and disease-related learning and
memory deficits. We examined the effects of EVOO on learning and memory
in SAMP8 mice, an age-related learning/memory impairment model
associated with increased amyloid- protein and brain oxidative damage.
We administered EVOO, coconut oil, or butter to 11 month old SAMP8
mice for 6 weeks. Mice were tested in T-maze foot shock avoidance
and one-trial novel object recognition with a 24 h delay. Mice which
received EVOO had improved acquisition in the T-maze and spent
more time with the novel object in one-trial novel object recognition
versus mice which received coconut oil or butter. Mice that received
EVOO had improve T-maze retention compared to the mice that received
butter. EVOO increased brain glutathione levels suggesting reduced
oxidative stress as a possible mechanism. These effects plus increased
glutathione reductase activity, superoxide dismutase activity, and
decreased tissue levels of 4-hydroxynoneal and 3-nitrotyrosine were
enhanced with enriched EVOO (3× and 5× polyphenols concentration).
Our findings suggest that EVOO has beneficial effects on learning
and memory deficits found in aging and diseases, such as those related
to the overproduction of amyloid- protein, by reversing oxidative damage
in the brain, effectsthat are augmented with increasing concentrations
of polyphenols in EVOO.

Synthetic analogues of flavonoids with improved activity against platelet activation
and aggregation as novel prototypes of food supplements
S Del Turco, S Sartini, G Cigni, C Sentieri, S Sbrana, et al.
Food Chemistry 175 (2015) 494–499 http://dx.doi.org/10.1016/j.foodchem.2014.12.005

We investigated the ability of quercetin and apigenin to modulate platelet activation
and aggregation, and compared the observed efficacy with that displayed by their
synthetic analogues 2-phenyl-4H-pyrido[1,2-a]pyrimidin-4-ones, 1–4, and 2,3-
diphenyl-4H-pyrido[1,2-a]pyrimidin-4-ones, 5–7. Platelet aggregation was
explored through a spectrophotometric assay on platelet-rich plasma (PRP)
treated with the thromboxane A2 mimetic U46619, collagen and thrombin in
presence/absence of various bioisosteres of flavonoids (12.5–25–50–100 lM).
The platelet density, (mean platelet component, MPC), was measured by the
Advia 120 Hematology System as a marker surrogate of platelet activation. The
induced P-selectin expression, which reflects platelet degranulation/activation,
was quantified by flow cytometry on PRP. Our synthetic compounds modulated
significantly both platelet activation and aggregation, thus turning out to be more
effective than the analogues quercetin and apigenin when tested at a
concentration fully consistent with their use in vivo. Accordingly, they might
be used as food supplements to increase the efficacy of natural flavonoids.

Polysaccharide Extracts From Sargassum Siliquosum J.G. Agardh Modulates
Production Of Pro-Inflammatory Cytokines In Lps-Induced Pbmc And Delays
Coagulation Time In-Vitro
RD Vasquez, RSP Garcia-Meim and JDA Ramos
Jour. Harmo. Res. Pharm., 2014, 3(3), 101-112  www.johronline.com

Sulfated polysaccharides from brown seaweeds exhibit various biological activities,
structural diversity, and are potential reagents for the development of therapeutic
drugs. This study aimed to determine the effect of aqueous and fucoidan extracts from
Sargassum siliquosum J. G. Agardh on viability of peripheral blood mononuclear
cells, production of pro-inflammatory cytokines and plasma coagulation using
in vitro
assays. Sulfate contents of the polysaccharides were quantified using Acid-Ashing Digestion Ion chromatography. Effect on viability of the extracts on
peripheral blood mononuclear cells was determined by MTT Assay. Estimation
of pro-inflammatory cytokines concentrations was done through Enzyme-Linked
Immunosorbent Assay, while anticoagulant activity was measured by Prothrombin
Time and Activated Partial Thromboplastin Time. Results revealed that both
extracts were non-cytotoxic to PBMCs, reduced significantly the production of
IL-1, IL-6,TNF-α and exhibited normal anticoagulant activity in PT assays and
prolonged APTT remarkably in dose-dependent manner. In conclusion, extracts
of the Sargassum siliquosum J.G. Agardh is a potential alternative source in
producing anti-inflammatory and anticoagulant substances in the future.

Purple corn anthocyanins inhibit diabetes-associated glomerular monocyte
activation and macrophage infiltration
Min-Kyung Kang, J Li, Jung-Lye Kim, Ju-Hyun Gong, Su-Nam Kwak, JHY Park, et al.
Am J Physiol Renal Physiol 303: F1060–F1069

Purple corn anthocyanins inhibit diabetes-associated glomerular monocyte activation
and macrophage infiltration. Diabetic nephropathy  (DN) is one of the major diabetic
complications and the leading cause of end- stage renal disease. In early DN, renal
injury and macrophage accumulation take place in the pathological environment
of glomerular vessels adjacent to renal mesangial cells expressing proinflammatory
mediators. Purple corn utilized as a daily food is rich in anthocyanins exerting
disease-preventive activities as a functional food. This study elucidated whether
anthocyanin-rich purple corn extract (PCA) could suppress monocyte activation and
macrophage infiltration. In the in vitro study, human endothelial cells and THP-1 monocytes were cultured in conditioned media of human mesangial cells exposed
to 33 mM glucose (HG-HRMC). PCA decreased the HG-HRMC-conditioned, media-induced expression of endothelial vascular cell adhesion molecule-1, E-selectin,
and monocyte integrins- and -2 through blocking the mesangial Tyk2 pathway. In the
in vivo animal study, db/db mice were treated with 10 mg/kg PCA daily for 8 wk. PCA
attenuated CXCR2 induction and the activation of Tyk2 and STAT1/3 in db/db mice.
Periodic acid-Schiff staining showed that PCA alleviated mesangial expansion-elicited renal injury in diabetic kidneys. In glomeruli, PCA attenuated the induction
of intracellular cell adhesion molecule-1 and CD11b. PCA diminished monocyte
chemoattractant protein-1 expression and macrophage inflammatory protein 2
transcription in the diabetic kidney, inhibiting the induction of the macrophage
markers CD68 and F4/80. These results demonstrate that PCA antagonized
the infiltration and accumulation of macrophages in diabetic kidneys through
disturbing the mesangial IL-8-Tyk-STAT signaling pathway. Therefore, PCA may
be a potential renoprotective agent treating diabetes-associated glomerulosclerosis.

Proximate analysis, phytochemical screening, and total phenolic and flavonoid
contentof Philippine bamboo Schizostachyum lumampao
JVV Tongco, RM Aguda and RA Razal.
Journal of Chemical and Pharmaceutical Research, 2014, 6(1):709-713

In Asia, bamboo has been widely cultivated as a fast growing non-timber forest
species. Flavonoids and phenolics were shown to reduce inflammation, promote
overall cardiovascular health and circulation, and even protect against certain kinds
of cancer. These studies necessitate the chemical characterization (e.g., proximate
analysis) and qualitative identification of phenolics.

The chemical composition of the leaves of Schizostachyum lumampao, known as
“buho” in the Philippines, was determined for its potential use as herbal tea with
potential health benefits, such as antioxidant properties. Proximate analysis using
standard AOAC methods showed that the air-dried leaves contain 10 % moisture, 30.5 % ash, 22.1 % crude protein, 1.6 % crude
fat, 28.7 % crude fiber, and 7.2 % total sugar (by difference). Using a variety of
reagents for qualitative phytochemical screening, saponins, diterpenes, triterpenes,
phenols, tannins, and flavonoids were detected in both the ethanolic and aqueous
leaf extracts, while phytosterols were only detected in the ethanolic extract. Using
UV-Vis spectrophotometry, the total phenolic content (in GAE) were 76.7 and
13.5 gallic acid equivalents per 100 g air-dried sample for the ethanolic and
aqueous extracts, respectively. The total flavonoid content were 70.2 and 17.86 mg
quercetin equivalents per 100 g air-dried sample for the ethanolic and aqueous
extracts, respectively. This preliminary study showed the total amount of phenolics
and flavonoids present in buho, the phytochemicals present, and its proximate

Ophiopogonin D: A new herbal agent against osteoporosis
Q Huang, B Gao, L Wang, Hong-Yang Zhang, Xiao-Jie Li, J Shi, Z Wang, et al.
Bone 74 (2015) 18–28

Excessive reactive oxygen species (ROS) play an important role in the development
of osteoporosis. Ophiopogonin D (OP-D), isolated from the traditional Chinese
herbal agent Radix Ophiopogon japonicus, is a potent anti-oxidative agent. We
hypothesized that OP-D demonstrates anti-osteoporosis effects via decreasing
ROS generation in mouse pre-osteoblast cell line MC3T3-E1 subclone 4 cells
and a macrophage cell line RAW264.7 cells. We investigated OP-D on osteogenic
and osteoclastic differentiation under oxidative status. Hydrogen peroxide (H2O2)
was used to establish an oxidative damage model. In vivo, we established a murine
ovariectomized (OVX) osteoporosis model. Then, we searched the molecular
mechanism of OP-D against osteoporosis. Our results revealed that OP-D
significantly promoted the proliferation of MC3T3-E1 cells and improved some
osteogenic markers. Moreover, OP-D reduced TRAP activity and the mRNA
expressions of osteoclastic genes in RAW264.7 cells. OP-D suppressed ROS
generation in both MC3T3-E1 and RAW264.7 cells. OP-D treatment reduced
the activity of serum bone degradation markers, including CTX-1 and TRAP.
Further research showed that OP-D displayed anti-osteoporosis effects via
reducing ROS through the FoxO3a-β-catenin signaling pathway. In summary,
our results indicated that the protective effects of OP-D against osteoporosis
are linked to a reduction in oxidative stress via the FoxO3a-β-catenin signaling
pathway, suggesting that OP-D may be a beneficial herbal agent in bone-related
disorders, such as osteoporosis.

Revealing the macromolecular targets of complex natural products
D Reker, AM Perna, T Rodrigues, P Schneider, M Reutlinger, et al.
Nature Chemistry Dec  2014; 6: 1072 – 1078

Natural products have long been a source of useful biological activity for the
development of new drugs. Their macromolecular targets are, however, largely
unknown, which hampers rational drug design and optimization. Here we present
the development and experimental validation of a computational method for the
discovery of such targets. The technique does not require three-dimensional
target models and may be applied to structurally complex natural products. The
algorithm dissects the natural products into fragments and infers potential
pharmacological targets by comparing the fragments to synthetic reference drugs
with known targets. We demonstrate that this approach results in confident
predictions. In a prospective validation, we show that fragments of the potent
antitumour agent archazolid A, a macrolide from the myxobacterium Archangium
gephyra, contain relevant information regarding its polypharmacology.
Biochemical and biophysical evaluation confirmed the predictions. The results
obtained corroborate the practical applicability of the computational approach to
natural product ‘de-orphaning’.

In vitro activity of Inula helenium against clinical Staphylococcus aureus strains
including MRSA
O’Shea S, Lucey B, Cotter L.
Br J Biomed Sci. 2009;66(4):186-9.

The present study aims to investigate the bactericidal activity (specifically
antistaphylococcal) of Inula helenium. The antimicrobial activity of the extract is
tested against 200 clinically significant Irish Staphylococcus aureus isolates
consisting of methicillin-resistant (MRSA) and -sensitive (MSSA) S. aureus
using a drop test method and a microbroth dilution method. The antibacterial
effect is evaluated by measuring the area of the inhibition zone against the
isolates. Results proved I. helenium to be 100% effective against the 200
staphylococci tested, with 93% of isolates falling within the ++ and +++ groups.
The minimum bactericidal concentration of I. helenium was examined on a subset
of isolates and values ranged from 0.9 mg/mL to 9.0 mg/mL. The extract was
equally effective against antibiotic-resistant and -sensitive strains. This plant
therefore possesses compounds with potent antistaphylococcal properties, which
in the future could be used to complement infection control policies and prevent
staphylococcal infection and carriage. This research supports other studies
wherein herbal plants exhibiting medicinal properties are being examined to
overcome the problems of antibiotic resistance and to offer alternatives in the
treatment and control of infectious diseases.

Inhibition of Proliferation of Breast Cancer Cells MCF7 and MDA-MB-231 by Lipophilic Extracts of Papaya (Carica papaya L. var. Maradol) Fruit
LE Gayosso-García Sancho, EM Yahia, P García-Solís, GA González-Aguilar
Food and Nutrition Sciences, 2014, 5, 2097-2103

Several epidemiological studies have suggested that carotenoids have
antineoplasic activities. The objective of this study was to determine the
antiproliferative effect of rich carotenoid lipophilic extracts of papaya fruit
pulp (Carica papaya L., cv Maradol) in breast cancer cells, MCF-7 (estrogen
receptor positive) and MDA-MB-231 (estrogen receptor negative), and in
non-tumoral mammary epithelial cells MCF-12F. Antiproliferative effect
was evaluated using the methyl-thiazolydiphenyl-tetrazolium bromide
(MTT) assay and testing lipophilic extracts from different papaya fruit
ripening stages (RS1, RS2, RS3, RS4), at different times (24, 48 and
72 h). Papaya lipophilic extracts do not inhibit cell proliferation of MCF-12F
and MDA-MB-231 cells. However, MCF-7 cells showed a significant
reduction in proliferation at 72 h with the RS4 papaya extract. Results
suggested that lipophilic extracts had different action mechanisms on
each type of cells and therefore, more studies were required to elucidate
such mechanisms.

In vitro cytotoxic activity of silver nano particle biosynthesized from Colpomenia
sinuosa and Halymenia poryphyroides using DLA and EAC cell lines
Vishnu Kiran M and Murugesan S
World J Pharm Sci 2014; 2(9): 926-930.

This study was conducted to investigate the invitro cytotoxic activity of silver
nanoparticles biosynthesized

from Colpomenia sinuosa and Halymenia poryphyroides using DLA and EAC
cell lines by tryphan blue dye  exclusion technique and MTT assay using Mouse L929 cell lines (Lungs fibroblast). The results of the trypan blue dye exclusion assay indicates that the silver nano particles biosynthesized from
Colpomenia sinuosa and Halymenia poryphyroides inhibits the growth of DLA
and EAC cell lines in a dose dependent manner against the standard drug
Curcumin where the silver nano particle biosynthesized from Colpomenia sinuosa
showed 61.57 % and silver nano particle biosynthesized from Halymenia poryphyroides showed 89.36 % in DLA cell line similarly the silver nanoparticle biosynthesized
from Colpomenia sinuosa showed 81.96 % and silver nanoparticle biosynthesized
from Halymenia poryphyroides 91.45 % in EAC cell line. The results of the MTT
assay indicated the silver nanoparticles biosynthesized from Colpomenia sinuosa
and Halymenia poryphyroides significantly inhibited the proliferation of L929 cells
in dose dependent manner where the silver nanoparticle biosynthesized from
Colpomenia sinuosa showed 37.06 % and silver nanoparticle biosynthesized from
Halymenia poryphyroides showed 100 % against the standard drug Curcumin.

Garlic compound fights source of food-borne illness better than antibiotics
·Better than antibiotics: Garlic compound fights source of food-borne illness

Researchers at Washington State University have found that a compound in garlic
is 100 times more effective than two popular antibiotics at fighting the Campylobacter
bacterium, one of the most common causes of intestinal illness. Their work was
recently published in the Journal of Antimicrobial Chemotherapy.  The discovery
opens the door to new treatments for raw and processed meats and food preparation
surfaces. Most infections stem from eating raw or undercooked poultry or foods
that have been cross-contaminated via surfaces or utensils used to prepare poultry.

Lu and his colleagues looked at the ability of the garlic-derived compound, diallyl
sulfide, to kill the bacterium when it is protected by a slimy biofilm that makes it
,000 times more resistant to antibiotics than the free floating bacterial cell. They
found the compound can easily penetrate the protective biofilm and kill bacterial
cells by combining with a sulfur-containing enzyme, subsequently changing
the enzyme’s function and effectively shutting down cell metabolism. The
researchers found the diallyl sulfide was as effective as 100 times as much
of the antibiotics erythromycin and ciprofloxacin and would often work in a
fraction of the time.

Two previous works published last year by Lu and WSU colleagues in Applied
and Environmental Microbiology and Analytical Chemistry found diallyl sulfide
and other organosulfur compounds effectively kill important foodborne pathogens,
such as Listeria monocytogenes and Escherichia coli O157:H7.

“Diallyl sulfide could make many foods safer to eat”, says Barbara Rasco, a
co-author on all three recent papers and Lu’s advisor for his doctorate in food
science. “It can be used to clean food preparation surfaces and as a preservative
in packaged foods like potato and pasta salads, coleslaw and deli meats”.

Effect of tree nuts on metabolic syndrome criteria: a systematic review and
meta-analysis of randomized controlled trials

SB Mejia, CWC Kendall, E Viguiliouk, LS Augustin, V Ha, AI Cozma, A Mirrahimi, et al.
BMJ Open 2014;4:e004660.  http://dx.doi.org:/10.1136/bmjopen-2013-004660

Objective: To provide a broader evidence summary to inform dietary guidelines of the
effect of tree nuts on criteria of the metabolic syndrome (MetS).
Design: We conducted a systematic review and metaanalysis of the effect of
tree nuts on criteria of the MetS.
Data sources: We searched MEDLINE, EMBASE, CINAHL and the Cochrane Library
(through 4 April 2014).
Eligibility criteria for selecting studies: We included relevant randomized controlled
trials (RCTs) of ≥3 weeks reporting at least one criterion of the MetS.
Data extraction: Two or more independent reviewers extracted all relevant data. Data
were pooled using the generic inverse variance method using random effects models
and expressed as mean differences (MD) with 95% CIs. Heterogeneity was assessed
by the Cochran Q statistic and quantified by the I2 statistic. Study quality and risk of
bias were assessed.
Results: Eligibility criteria were met by 49 RCTs including 2226 participants who
were otherwise healthy or had dyslipidemia, MetS or type 2 diabetes mellitus.
Tree nut interventions lowered triglycerides (MD=−0.06 mmol/L (95% CI −0.09
to −0.03 mmol/L)) and fasting blood glucose (MD=−0.08 mmol/L (95% CI −0.16
to −0.01 mmol/L)) compared with control diet interventions. There was no effect
on waist circumference, high-density lipoprotein cholesterol or blood pressure with
the direction of effect favoring tree nuts for waist circumference. There was
evidence of significant unexplained heterogeneity in all analyses (p<0.05).
Conclusions: Pooled analyses show a MetS benefit of tree nuts through modest
decreases in triglycerides and fasting blood glucose with no adverse effects
on other criteria across nut types. As our conclusions are limited by the short
duration and poor quality of the majority of trials, as well as significant
unexplained between-study heterogeneity, there remains a need for larger,
longer, high-quality trials.

DPPH free radical scavenging activity of phenolics and flavonoids in some medicinal
plants of India
R Patel, Y Patel, P Kunjadia and A Kunjadia
Int.J.Curr.Microbiol.App.Sci (2015) 4(1): 773-780 http://www.ijcmas.com

Methanolic extracts of Gymnema sylvestre (leaf), Holarrhena antidysenterica (bark),
Vernonia anthelmintica(seeds) Enicostemma littorale (leaf), Momordica charantia
(fruit), Swertia chirata (leaf), Azadirachta indica (leaf), Caesalpinia bonducella (leaf)
used in Ayurvedic medicines for number of ailments were evaluated for their
antioxidant activity.The free radical-scavenging activity of the extracts was measured
as decolorizing activity followed by the trapping of the unpaired electron by 1, 1-
diphenyl-2-picryl hydrazyl radical (DPPH). The percentage decrease of DPPH
was recorded maximum in A. indica followed by M. charantia, C. bonducella,
E.littorale, V. anthelmintica, S.chirata, H.antidysenterica, G.sylvestre. The
antioxidant activity of medicinal plants was at par with the commercial antioxidant
like L-Ascorbic acid. Phytochemical analysis revealed the presence of major
phytocompounds like terpenoids, alkaloids, glycosides, phenolics and tannins.
Moreover, total flavonoid concentration equivalents to gallic acid was found in
the range of 326 μg to 1481μg/g of plant extracts and that of total phenolic
concentration equivalents to phenol was found in the range of 23.50 μg to
89.82 μg/g of plant extracts. The findings indicated promising antioxidant
activity of crude extracts of the above plants and needs further exploration
for their effective use in both modern and traditional system of medicines.

Cyanobacterial natural products as antimicrobial agents
V.D. Pandey
Int.J.Curr.Microbiol.App.Sci (2015) 4(1): 310-317 http://www.ijcmas.com

Cyanobacteria (blue-green algae) constitute a morphologically diverse and
widely distributed group of Gram-negative photosynthetic prokaryotes. Possessing
tremendous adaptability to varying environmental conditions, effective protective
mechanisms against various abiotic stresses and metabolic versatility, they colonize
and grow in different types of terrestrial and aquatic habitats. In addition to
the potential applications of cyanobacteria in various fields, such as agriculture,
aquaculture, pollution control, bioenergy and nutraceuticals, they produce chemically
diverse and pharmacologically important novel bioactive compounds, including
antimicrobial compounds (antibacterial, antifungal and antiviral). The emergence
and spread of antibiotic resistance in pathogenic microbes against commonly used
antibiotics necessitated the search for new antimicrobial agents from sources other
than the traditional microbial sources (streptomycetes and fungi). Various features
of cyanobacteria, including their capability of producing antimicrobial compounds,
make them suitable candidates for their exploitation as a natural source
of antimicrobial agents.
Determination of nutritional value and antioxidant from bulbs of different onion
(Allium cepa) variety: A comparative study
Kandoliya, U.K.*, Bodar, N.P., Bajaniya, V.K., Bhadja N.V. and Golakiya, B.A.
Int.J.Curr.Microbiol.App.Sci (2015) 4(1): 635-641 http://www.ijcmas.com

Onion (Allium cepa) is one of the most economically important vegetable crops
consumed for their ability to enhance the added flavor and typical taste in other
foods. It is a good source of antioxidants as well as some phytonutrients.
So the experiment was conducted to study the nutritional quality along with
various parameters contributing antioxidant activity from onion of different red and
white type local varieties. The findings revealed from all the variety studied,
shows 58.14 to 77.67 % DPPH value, comparable amount of flavanoids
(0.422 to 1.232 mg.g-1) and anthocyanine content along with total phenol
(8.96-18.23 mg.100 g-1), Pyruvic acid (1.09 to 1.33 mg.g-1), ascorbic acid
(1.18 to 3.89 mg.100g-1) , protein (0.79 to 1.27%) and titrable acidity
(0.34 0.75%).These results reveal that JDRO-07-13 of Red variety and
GWO-1 of white nutritionally found better due to its higher antioxidant
property, proteins, carbohydrates, reducing sugar and should be included in diets to supplement our daily allowance needed by the body.

Curcumin: New Weapon against Cancer
Fayez Hamam
Food and Nutrition Sciences, 2014, 5, 2257-2264

All the evidences point out to the fact that the incidence, mortality and number of
persons living with cancer are on the rise and, thus, this will impose a significant
burden on health care resources. The considerable number of deaths from cancer
necessitates the need to developing novel alternative cures that are efficient, safe,
cheap and easy to use. In the search for new therapies for tumors, naturally-derived compounds have been considered as a good source of novel anticancer
drugs. The challenge here is to find products that are pharmacologically active
against tumor cells with suitable toxicity profile and least damage to normal cells.
Curcumin is a spice widely used in many countries especially in South Asia and
it has gained importance for its anticancer function and low toxicity toward normal
tissues in a range of biological systems. In spite of significant research works, many
difficulties hinder its oral use in the therapy of different kind of tumors, such as
extreme low solubility in water, quick break down and excretion after being absorbed
in the human body. Low bioavailability due to enhanced metabolism and rapid
system elimination is another problem that hinders oral use of curcumin as
anticancer agent. Therefore, the previously mentioned poor pharmacokinetics
characteristics inhibit curcumin from reaching its site of action and, thus,
lessen its effectiveness against tumors. This article reviews the latest global
cancer statistics with special attention to be directed toward ovarian cancer.
It sheds light on many research works that investigated the protective and
therapeutic functions of different curcumin preparations against different
sites of cancer using animal models. It also summarizes recent
research works concerning the antitumor effects of curcumin alone and/or
loaded into a range of delivery devices in many types of ovarian cancer cell lines.

Cinnamon is lethal weapon against E. coli O157:H7

When cinnamon is in, Escherichia coli O157:H7 is out.  That’s what researchers
at Kansas State University discovered in laboratory tests with cinnamon and
apple juice heavily tainted with the bacteria.  Presented at the Institute of Food
Technologists’ 1999 Annual Meeting in Chicago on July 27, the study findings
revealed that cinnamon is a lethal weapon against  E. coli O157:H7 and may be
able to help control it in unpasteurized juices.

Lead researcher Erdogan Ceylan, M.S., reported that in apple juice samples
inoculated with about one million E. coli O157:H7 bacteria, about one teaspoon
(0.3 percent) of cinnamon killed 99.5 percent of the bacteria in three days at room
temperature (25 C).  When the same amount of cinnamon was combined with
either 0.1 percent sodium benzoate or potassium sorbate, preservatives approved
by the Food and Drug Administration, the E. coli were knocked out to an
undetectable level.  The number of bacteria added to the test samples was
100 times the number typically found in contaminated food.

“If cinnamon can knock out E. coli O157:H7, one of the most virulent foodborne
microorganisms that exists today, it will certainly have antimicrobial effects on other
common foodborne bacteria, such as Salmonella and Campylobacter,” noted Daniel
Y.C. Fung, Ph.D., professor of Food Science in the Department of Animal Sciences
and Industry at K-State, who oversaw the research.

Last year, Fung and Ceylan researched the antimicrobial effects of various spices
on  E. coli O157:H7 in raw ground beef and sausage and found that cinnamon,
clove, and garlic were the most powerful.  This research led to their recent studies
on cinnamon in apple juice, which proved to be a more effective medium than meat
for the spice to kill the bacteria.

“In liquid, the E. coli have nowhere to hide,” Fung noted, “whereas in a solid structure,
such as ground meat, the bacteria can get trapped in the fat or other cells and
avoid contact with the cinnamon.  But this cannot happen in a free-moving environment.”

For a copy of the study presented at IFT’s Annual Meeting, contact Angela Dansby at
312-82-8424 x127 or via e-mail at aldansby@ift.org
Anti-inflammatory, anti-proliferative and anti-atherosclerotic effects of quercetin in
human in vitro and in vivo models
R Kleemann, Lars Verschuren, M Morrison, S Zadelaar, MJ van Erk, PY Wielinga, & T  Kooistra
Atherosclerosis 218 (2011) 44– 52

Objective: Polyphenols such as quercetin may exert several beneficial effects,
including those resulting from anti-inflammatory activities, but their impact on
cardiovascular health is debated. We investigated the effect of quercetin on
cardiovascular risk markers including human C-reactive protein (CRP) and on
atherosclerosis using transgenic humanized models of cardiovascular disease.
Methods: After evaluating its anti-oxidative and anti-inflammatory effects in
cultured human cells, quercetin (0.1%, w/w in diet) was given to human CRP
transgenic mice, a humanized inflammation model, and ApoE*3Leiden transgenic
mice, a humanized atherosclerosis model. Sodium salicylate was used as an
anti-inflammatory reference. Results: In cultured human endothelial cells,
quercetin protected against H2O2-induced lipid peroxidation and reduced the
cytokine-induced cell-surface expression of VCAM-1 and E-selectin. Quercetin
also reduced the transcriptional activity of NFB in human hepatocytes. In human
CRP transgenic mice (quercetin plasma concentration: 12.9 ± 1.3 M), quercetin
quenched IL1-induced CRP expression, as did sodium salicylate. In ApoE*3 Leiden mice, quercetin (plasma concentration: 19.3 ± 8.3 M) significantly attenuated
atherosclerosis by 40% (sodium salicylate by 86%). Quercetin did not affect
atherogenic plasma lipids or lipoproteins but it significantly lowered the circulating
inflammatory risk factors SAA and fibrinogen. Combined histological and microarray
analysis of aortas revealed that quercetin affected vascular cell proliferation thereby
reducing atherosclerotic lesion growth. Quercetin also reduced the gene expression
of specific factors implicated in local vascular inflammation including IL-1R, Ccl8, IKK,
and STAT3.
Conclusion: Quercetin reduces the expression of human CRP and cardiovascular risk
factors (SAA, fibrinogen) in mice in vivo. These systemic effects together with local
anti-proliferative and anti-inflammatory effects in the aorta may contribute to the
attenuation of atherosclerosis.
Natural products to drugs: natural product derived compounds in clinical trials
Mark S. Butler
Nat  Prod  Rep  2005; 22 : 162 – 195 http://dx.doi.org:/10.1039/b402985m

Natural product and natural product-derived compounds that are being
evaluated in clinical trials or in registration (current 31 December 2004)
have been reviewed. Natural product derived drugs launched in the
United States of America, Europe and Japan since 1998 and new
natural product templates discovered since 1990 are discussed.

Natural Products (NPs) traditionally have played an important role in drug discovery
and were the basis of most early medicines. Over the last 10 to 15 years advances
in X-ray crystallography and NMR, and alternative drug discovery methods such as
rational drug design and combinatorial chemistry have placed great pressure upon
NP drug discovery programs and during this period most major pharmaceutical
companies have terminated or considerably scaled down their NP operations.
However, despite the promise of these alternative drug discovery methods, there is
still a shortage of lead compounds progressing into clinical trials. This is especially
the case in therapeutic areas such as oncology, immunosuppression and metabolic
diseases where NPs have played a central role in lead discovery. In a recent review,
Newman,Cragg and Snader analysed the number of NP-derived drugs present in
the total drug launches from 1981 to 2002 and found that NPs were a significant
source of these new drugs, especially in the oncological and antihypertensive
therapeutic areas. In addition to providing many new drug leads, NPs and NP-derived drugs were well represented in the top 35 worldwide selling ethical drugs
in 2000, 2001 and 2002.

Antibacterial activity of green tea (Camellia sinensis) Extract against dental
caries and other pathogens
P. Lavanya and M. Sri priya
Int.J.Adv. Res.Biol.Sci.2014; 1(5):58-70

The present study has however, revealed that the herbal plant Camellia sinensis (green tea) possess antimicrobial properties. The isolated strains were confirmed by performing staining and biochemical techniques. Aqueous extract of green tea were taken and used for the study of inhibition effect against dental caries and
other pathogens. The zone of inhibition was performed using agar well diffusion techniques different concentration of green tea extracts were studied for their
antibacterial activity. The overall results showed that the microorganisms
were susceptible to different concentration of aqueous extracts of Camellia
sinensis which is a function of their antimicrobial properties. The effectiveness of active principle was studied and compared with the previous one. The nature
of the chemicals present as active principle of the extract was studied using
Paper chromatography and Thin layer chromatography. The chemicals involved in
antimicrobial activity are commonly belonging to any one of the group such as flavanoids, alkaloids, saponins and polyphenols. It could be concluded
that flavonoid in a potential natural, antimicrobial agent against dental
caries and other pathogens.

Antibacterial activity of Mangrove Medicinal Plants against Gram positive
Bacterial pathogens
K. A. Selvam* and K. Kolanjinathan
Int. J. Adv. Res. Biol.Sci. 1(8): (2014): 234–241

Ten mangrove medicinal plants viz., Avicennia marina, Rhizophora mucuronata, Rhizophora mangle, Asparagus officinalis, Ceriops decandra, Aegiceras
corniculatum, Acanthus ilicifolius, Bruguiera cylindrica, Rhizophora apiculata and Xylocarpus grantum were collected from mangrove forest of Pichavaram, Tamil
Nadu, India. The antibacterial activity of mangrove plant extracts (150 mg/ml and
300 mg/ml) were determined by Disc diffusion method. The zone of inhibition was more at 300 mg/ml of extracts when compared to 150 mg/ml of extracts. The
antibacterial activity of selected mangrove plant leaf extracts was determined
against pathogenic bacterial isolates. The methanol extract of Ceriops decandra showed maximum zone of inhibition against all the bacterial isolates followed
by Avicennia marina, Rhizophora mucronata, Aegiceras corniculatum, Rhizophora apiculata, Rhizophora mangle, Acanthus ilicifolius, Asparagus officinalis, Xylocarpus grantum and Bruguiera cylindrica at 300 mg/ml. The hexane extract of mangrove plants showed minimum inhibition zone against bacterial pathogens
when compared to the other solvent extracts. The DMSO was used as a blind
control and the antibiotic Ampicillin (300 mg/ml) was used as a positive control. Minimum inhibitory concentration (MIC) of the mangrove plant extracts against bacterial isolates was tested in Mueller Hinton broth by Broth macro dilution
method. The MIC of mangrove plants against bacterial pathogens was ranged
between 20 mg/ml to 640 mg/ml.

Antioxidant and antibacterial activity of Berberis tinctoria root
Karthikkumar Va, Sharanya R , Allegendiran R, Sasikumar J.M
Int. J. Adv. Res. Biol.Sci. 1(9): (2014): 292–297
Herbs have always been the principle form of medicine in developing nations
and presently they are becoming popular throughout the developed world as
people strive to stay healthy in the face of chronic stress and to treat illness with medicines that work in concert with body’s own defences. The aim of the present study was to evaluate the antioxidant and antibacterial potential of Bereris
tinctoria root. Plant material collected and extracted with various solvents. Different concentrations of extracts were used to evaluate the potential. Bereberis tinctoria
root at a concentration of 1000μg/ml shows high antioxidant activity and relatively
all extracts possessing strong to moderate antibacterial activity. In addition, during phytochemical screening, we got saponins and sterols from its root, when extracting with organic solvents. Thus, root extract of Berberis tinctoria might be good
candidate for the synthesis of antibacterial drugs in the future.

Biological Activities of Soybean Galactomannan Oligosaccharides and
Their Sulfated Derivatives
MMI Helal, SA Ismail, MOI Ghobashy, SS Elgazar, et al.
Int.J.Adv. Res.Biol.Sci.2014; 1(6):113-121

Galactomanno-oligosaccharieds (GMO) and their sulfated derivatives
(SGMO) were prepared from soybean hulls and evaluated for their biological
activities as anticoagulant; antimicrobial; antitumor; fibrinolytic and prebiotics.
The results indicated that the sulfating process has positive effect on the
anticoagulation and fibrinolytic activities of the galactomanno-oligosaccharides.
The SGMO have prolonged clotting time more than 24h at concentration resemble that of the standard heparin. It was also found that the SGMO have fibrinolytic
activity as that of the standard hemoclar and 3 times higher than that of the native GMO oligosaccharides. The prepared oligosaccharides also preformed anti-tumor
activity against human colon carcinoma cell line and the percentage of the dead cells increase from 28% to 72% by increase the concentration of the oligosaccharides from 0.005 to 0.02 mg/ml. The tested galactomanno-oligosaccharides also act as good source for prebiotic as they have the ability to grow the beneficial bacteria
4 to 8 times higher than the pathogenic one. To our knowledge this is the first
time someone report anticoagulation; fibrinolytic and direct antitumor activities for galactomanno-oligosaccharides not to mention soybean galactomanno-oligosaccharides.

Biotechnological Application of Production β-Lactamase Inhibitory Protein
(BLIP) By Actinomycetes Isolates from Al-Khurmah Governorate
HM Atta;  RA Bayoumi and  MH El-Sehrawi
Int. J. Adv. Res. Biol.Sci. 1(7): (2014): 144–154

Many pathogenic bacteria secrete β-lactamase enzymes as a mechanism of
defense against β-lactam antibiotics. Sixty-nine unrepeated actinomycetes
isolates were isolated from different localities in Al-Khurmah governorate, Saudi Arabia kingdom. Actinomycetes isolates were screened for producing β-lactamase inhibitory effect against amoxicillin –resistant bacteria. There were eleven isolates (15.94 %) which had β-lactamase inhibitory protein (BLIP) effect against amoxicillin –resistant Staphylococcus aureus, pseudomonas aeruginosa and Klebsiella
pneumonia. The KH-3201-144 isolate has been considered the most potent, this
was identified by biochemical, chemotaxonomic, morphological and physiological properties consistent with classification in the genus Streptomyces, with the
nearest species being Streptomyces rimosus. Furthermore, a phylogenetic
analysis of the 16S rDNA gene sequence and ribosomal database project
consistent with conventional taxonomy confirmed that strain KH-3201-144
was most similar to Streptomyces rimosus (96%). The highest amount of
β-lactamase inhibitory protein was precipitated at 40% of saturated ammonium sulphate. The purification was carried out by using both diethyl-aminoethyl-cellulose G-25 and sephadex G-200 column chromatography, respectively.
The β-lactamase inhibitory protein was separated at 40 KDa. The minimum
inhibition concentrations “MICs” of the purified β-lactamase inhibitory protein
(BLIP) effect against amoxicillin –resistant Staphylococcus aureus, pseudomonas aeruginosa and Klebsiella pneumonia were also determined.

Bioactive compounds from marine Microbes
P.Sudhasupriya and M.Rajalakshmi
Int.J.Adv. Res.Biol.Sci.2014; 1(6):232-236

Natural compounds isolated from marine organisms have been found to be
a very rich source of bioactive molecules. Reported biological effects of these compounds include anti‐tumor, anti-inflammatory and anti‐viral activities as
well as immunomodulatory and analgesic properties. Pharmaceutical market is growing rapidly and continuously. But, still the demand for new drug discovery
is encouraged. The reason behind this motivation can be the growing number
of drug–resistant infectious diseases and more and more upcoming disorders. Pharmaceutical market is growing rapidly and continuously. But, still the demand
for new drug discovery is encouraged. The reason behind this motivation can
be the growing number of drug–resistant infectious diseases and more and more upcoming disorders.

The Discovery and Properties of Avemar – Fermented Wheat Germ
Extract: Carcinogenesis Suppressor
Larry H Bernstein, MD, FCAP, Contributor

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