Taurisolo®, a novel nutraceutical formulation based on grape pomace polyphenols, as a tool for the management of oxidative stress- and atherosclerosis-related diseases

The present PhD thesis summarises evidence from all the studies performed during the three-year PhD Course in Pharmaceutical Science, at the NutraPharmaLabs of the Department of Pharmacy, University of Naples Federico II, having as mail goal the evaluation of the nutraceutical potential of Taurisolo®, a novel nutraceutical formulation based on grape pomace polyphenolic extract. In particular, the present PhD project has been conducted in cooperation with both industrial (MBMed Company, Turin, Italy) and foreign University (University of Balearic Islands, Palma del Mallorca, Spain) partners. The entire project was planned with a dual view: design and formulate a novel nutraceutical product starting from re-use of agri-food by-products, following all the steps required by the nutraceutical industry, including preventive characterisation of the chemical profile, evaluation of bioaccessibility and bioavailability of bioactive compounds, optimisation of the productive processes and their translation in large-scale, marketing techniques evaluation of the biological activities, following a pharmacological approach including pre-clinical and clinical studies. More specifically, results herein presented refer to all the studies performed in our Labs and in collaboration with other Departments and Research Institutes. They include in vitro studies, animal-based studies and randomised clinical trials conducted on both healthy and pathological subjects. Further studies are still ongoing with the aim to clarify the main putative mechanisms of action for the cardioprotective role played by Taurisolo® or to provide novel insight regarding the observed clinical results. In summary, data so far collected allow concluding that Taurisolo® is a useful and valid polyphenol-based formulation with promising nutraceutical properties in reduction of risk factors related to both development and progression of cardiovascular diseases, in particular atherosclerosis

Oleuropein as a novel anti-diabetic nutraceutical.An overview

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Resveratrol-based Nutraceuticals for theManagement of Diabetes and Obesity: RealTherapeutic Potential or a mere Palliative?

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Intermediate Alpha-1 Antitrypsin Deficiency Can Play a Role in Pulmonary Exacerbation?

Alpha-1 antitrypsin deficiency is generally suspected in young patients with pulmonary emphysema or chronic obstructive pulmonary disease (COPD). Patients often suffer from diagnostic delays or are misdiagnosed, for example, with COPD, asthma, or airway hyperresponsiveness because of the nonspecific nature of respiratory symptoms recognised with Alpha-1 antitrypsin deficiency (AATD). These pathologies develop in homozygous patients (both compromised alleles) with severely deficient protein; however, they are also frequently observed in heterozygous patients (only one compromised allele) for the gene mutation with a more or less deficient protein and functional anatomical damage of varying severity depending on the type of mutation and the exposure to environmental risk factors and/or professional that can trigger the repeated injurious inflammatory process. Case Description: We describe two cases of late diagnosis of alpha- 1 antitrypsin deficiency, with many exacerbations and intermediate level of alpha-1 antitrypsin. Due to the peculiar clinical history, and the PLowell rare mutation, although intermediate AATD, the patients were subjected to replacement therapy and they obtained clinical improvement. Discussion: Both the cases carried a heterozygous PLowell mutation representing two interesting and rare examples of clinical cases with double heterozygosity. The presence in the other AAT allele of the S- mutation in the first case and a concomitant presence of another mutation in the cystic fibrosis gene in the second case contributed to the protease-antiprotease imbalance and, despite intermediate AATD, was the probable cause of the numerous exacerbations. Conclusion: Alpha-1 antitrypsin deficiency should always be suspected in patients with respiratory disease and an unclear or complex clinical history. It may be useful to recognize and evaluate treatment even outside the established parameters, in selected cases

Photodynamic therapy and adapalene in dermatology: synergistic effects in immortal human keratinocytes

We compared the effects of methyl levulinate (MAL)/photodynamic therapy (PDT) and adapalene, evaluated singly, versus combination therapy on HaCaT keratinocytes. Our aim was to determine whether the additive/synergistic outcomes of combination treatment were such that doses of each component could be reduced without affecting treatment efficacy. The analysis of data from specific PDT/adapalene combinations results indicating effects ranging from additive to clearly synergistic. We first evaluated the effects on cell viability, cell cycle and protein expression profiles of each individual treatment, then we tried to understand the reasons and the mechanisms whereby cells under combined treatment move more efficiently to death. Although we observed therapeutic strengthening by increasing either drug doses or light fluences, reinforcement was particularly marked in combinations in which PDT was predominant. Thus, if PDT is appropriately tuned, the dose of the drug can be reduced without compromising the therapeutic response. As both photodynamic therapy and adapalene have been and are therapeutic approaches to treat different dermatological pathologies such as acne, actinic keratosis etc., our data suggest that the adapalene-PDT combination may have important spin-off in clinical dermatology as a strategy to tackle this nasty condition

Expression of Tight Junction and Drug Efflux Transporter Proteins in an in vitro Model of Human Blood–Brain Barrier

Interendothelial cell tight junctions (TJs) proteins contribute to maintain the structural and functional integrity of the blood–brain barrier (BBB) and several efflux transporters regulate transport of compounds across BBB. A unique double compartment-model of the BBB, consisting of cerebral endothelial cells isolated from cryopreserved human glial tumors, alone and in the presence of human astroglial cells derived from the same tissue preparation was established. Endothelial cell viability and transendothelial electrical resistance (TEER) were measured in this model and three representative TJ proteins – occludin (OCLN), zonula occludens-1 (ZO-1) and claudin-5 (CLN-5) – as well as several drug efflux transporters – P-glycoprotein (P-gp), multidrug resistance protein-1 and 2 (MRP-1 and MRP-2), organic anion-transporting polypeptide-1 and 3 (oatp1 and oatp3) were analyzed at both the protein and gene transcript level. Functional activity of P-gp and MRP-1 was also assessed. Endothelial cell viability as well as TEER significantly increased in the presence of glial cells. A significant increase of expression of OCLN, ZO-1, and CLN-5 proteins as well as of several drug transporter proteins except oatp3 and MRP-1, was also found in the presence of glial cells. All the gene transcripts protein analyzed were found to be significantly increased in the presence of glial cells. A suitable functional activity of P-gp and MRP-1 was also found. These results demonstrate that this brain endothelium culture system mimics a physiologically relevant situation and may therefore provide a new tool for studying the effects of biological fluids such as serum and cerebrospinal fluid from patients with neurological disorders underlying a BBB alteration in disease pathogenesis

Novel Penicillin-Type Analogues Bearing a Variable Substituted 2-Azetidinone Ring at Position 6: Synthesis and Biological Evaluation

The synthesis and the biological activity of novel semi-synthetic β-lactam compounds containing an azetidinone moiety joined to the amino-nitrogen of the (+)-6-aminopenicillanic acid (6-APA) as new antibacterial agents is reported. The synthesized compounds were screened for their in vitro antimicrobial activity against a panel of Gram positive and Gram negative pathogens and environmental bacteria. Tested compounds displayed good antimicrobial activity against all tested Gram positive bacteria and for Staphylococcus aureus and Staphylococcus epidermidis antimicrobial activity resulted higher than that of the reference antibiotic. Additionally, in vitro cytotoxic screening was also carried out indicating that the compounds do not cause a cell vitality reduction effective at concentration next to and above those shown to be antimicrobial

Fohotodermatoses and Skin Cancer

Preface Skin cancer is one of the most common types of tumors in Western countries. In the United States only, more than one million people are diagnosed with skin cancer each year. Although the absolute number of skin cancer patients is increasing, the death is inversely decreasing, due to the early detection and treatment. Basal cell carcinoma (BCC), squamous cell carcinoma (SCC), and melanoma are three major types of skin cancer. BCC and SCC rarely have metastasis; over 95 percent BCC and SCC patients can be cured. Melanoma only accounts for a small percentage of skin cancer, but it causes 75 percent death of this disease. In this book, we invited a number of experts to present their latest accomplishments on skin cancer research. Although the topics are varied, the authors did great work to help readers better understand skin cancer and learn the knowledge to prevent this disease. There are three sections in this book, starting with etiology. Ultraviolet (UV) light exposure is overwhelmingly believed to be the most frequent cause of skin cancer. In this section, the association between UV and photodermatoses, as well as skin cancer is discussed. Desmosomal cadherins are important molecules in tumor cell adhesion and invasion, and their important roles in BCC are also presented in details. In the diagnosis and treatment section, a few new methodologies are described. As known, the outcome of malignant melanoma greatly depends on the thickness of the tumor at the time of treatment. Accurate determination of melanoma lateral and depth of margins using non-invasive imaging technologies is of importance when making sound decisions for treatment and evaluating a five year survival rate. A novel method named differential scanning calorimetry is capable of predicting metastasis of melanoma patients by monitoring the temperature changes of plasma. Electronic miniature X-ray brachytherapy is introduced as a new technology to treat nonmelanoms skin cancer. Although its potential has not yet been fully realized, chemoprevention, in terms of using chemical agents that naturally occur in foods, or are administered as pharmaceuticals to retard or reverse the process of carcinogenesis and progression of cancer, has been recognized to benefit individuals with precancerous lesions or genetic susceptibilities to cancer. In the prevention section, two chapters summarized the most recognized dietary phytochemicals and their potential application in skin cancer. X Preface This book would not have been possible without the contributions of all authors and the support from the publisher. Especially, I will convey my sincere appreciation to Ms. Tajana Jevtic, who has always been available and supportive of me to accomplish this project. Yaguang Xi, M.D., Ph.D. Assistant Professor of Oncologic Sciences, University of South Alabama, US

Stereoselective Reaction of 2-Carboxythioesters-1,3-dithiane with Nitroalkenes: An Organocatalytic Strategy for the Asymmetric Addition of a Glyoxylate Anion Equivalent.

Under mild reaction conditions γ-nitro-β-aryl-α-keto esters with up to 92% ee were obtained, realizing a formal catalytic stereoselective conjugate addition of the glyoxylate anion synthon