-2013 2014 - Wanda Rizza, Washington University

The scholarship holder's final report Wanda Rizza

The opportunity to go to Washington University in St. Louis (Department of Applied Physiology, Division of Geriatric and Nutritional Sciences) and to spend a period of twelve months (from February 2013 to February 2014), as intense as they were rewarding, allowed me to significantly enrich my scientific and didactic background, constituting a unique experience for my growth both professionally and personally. Furthermore, being the first experience of experimental research undertaken after obtaining the Master's Degree, it certainly represented an opportunity to confront myself with a working and relational reality, different from those known up to that moment.

Firstly, I had the opportunity to experience how teamwork is fundamental for the achievement of common objectives, for which we are committed with equal intensity regardless of the task assigned and the consequent load of responsibility; in the working context, every day takes place in a different way, which is why being able to optimize times, know how to deal with the unexpected and carry out one's tasks in synergy with those of the entire team become key factors for harmonizing the work of group.

The need to speak another language prompted me from the first moment to deepen my initial knowledge, to be able to make myself better understood by colleagues and patients, and this proved to be a very important aspect for better integration in the department.

I also had the opportunity to write, together with Principal Investigator Dr Luigi Fontana and his colleague Dr. Nicola Veronese, and publish a review article in Aging Research Reviews (“What are the roles of calorie restriction and diet quality in promoting healthy longevity?”).

During the research period I followed the following clinical studies:

• Vegan Study, a clinical trial whose objective is to evaluate the physiological, biochemical and Gene Expression effects, as well as the beneficial effects on health, of Calorie Restriction in men, by comparing three different types of diet: the common American diet (“Usual USA diet”) followed by sedentary patients but free from chronic pathologies; nutrition followed by physically active patients (“exercisers group”); the "Calorie Restricted Diet" (CR) based on the exclusion or strong reduction of typical foods in the modern diet, such as red meats, refined foods, foods rich in sugars and saturated fats, followed by the "Calorie Restricted" patients. The study in question also aims to obtain clinical evidence on the possible role of CR in the slow aging process in humans through the study of gene expression on sigmoid cells, a tissue with a high carcinogenic potential.
• Protein Study, clinical trial on the evaluation of the effect of a "Low Protein Diet" on the slowing down/inhibition of prostate tumor growth in patients who have to undergo surgery to remove the prostate (group 1) and in patients who have had a rise in PSA (prostate specific antigen) after surgery (group 2).
• Intermittent Fasting Study (started in August 2013), a clinical trial designed to determine the metabolic and cardiovascular effects of Intermittent Fasting (IF) in patients with BMI values ​​between 24 and 35 kg/m2, and to evaluate whether the same IF is able to reduce levels of chronic inflammation, monitoring the reduction of C-reactive protein (hsCRP) as the main reference outcome. The secondary objective of the study is to evaluate whether IF reproduces the metabolic, molecular and cardiovascular effects of Calorie Restriction, by monitoring weight and body composition, measuring hormonal parameters (insulin, IGF-1, IGFBP- 3, testosterone, estradiol, sex hormone binding globulin (SHGB), TSH, T3, leptin, adiponectin), and markers of oxidative stress, as well as lipid profile, glucose tolerance and blood pressure.

Methodologies (common to all clinical trials unless expressly specified):

• In-depth analysis of food diaries (4-day and 7-day food diaries) using NDSR (Nutrition Data System for Research) software, and subsequent storage and integration of the nutritional information obtained with other clinical data, collected concurrently by other team members research through special questionnaires and blood tests of patients.
• Drawing up of questionnaires regarding the eating habits of patients participating in the screening, to validate their eligibility for ongoing studies.
• ​Active participation in screening and randomization visits, and in intervention meetings and patient follow-up.
• ​Preparation of power point presentations to be shown to patients randomized to the intervention group of one of the studies under review (Intermittent Fasting Study).
• ​​Collection of bioptic samples of the colon mucosal tissue: in conjunction with the sigmoidoscopy procedure, performed by a gastroenterologist Doctor, sampling of small fractions of tissue, in number from 10 to 20 and 3 to 10 mm in size, and their conservation in the freezer at - 80 ◦C. This procedure takes place at the Clinical Research Unit, an operating unit in which all the clinical procedures to which patients are subjected take place.
• Assistance during the administration of OGTT (Oral Glucose Tolerance Test) to patients participating in the Intermittent Fasting Study.
• ​​Laboratory: isolation technique of PBMC (peripheral blood mononucleated cell) starting from a blood sample (in EDTA) of the patient. The above procedure involves the use of Ficoll, a hydrophilic polysaccharide which allows the blood to be separated into layers, creating an upper layer of plasma, followed by a layer of PBMCs and a basal fraction of polymorphonuclear cells and erythrocytes. The PBMCs are subsequently extracted through a series of washings and centrifugations, counted by cell counting technique, sampled and stored by freezing at -80 ◦C.​
• Bibliographic research on topics related to the clinical studies followed.

Preliminary results:

• Vegan Study: the study involves carrying out genetic tests which will take place in the months following the conclusion of the same.
• Protein Study: table comparing PSA values ​​(baseline and after one month of follow-up) between n=12 patients and n=12 controls.
• Intermittent Fasting Study: the clinical trial started a few months ago, therefore it will be possible to draw the first conclusions, and compare the data with those obtained at the Baseline stage, upon reaching six months of participation for a certain number of patients. However, the first data in our possession show that IF leads to a significant reduction in body weight compared to baseline.