Free-amino acid metabolic profiling of visceral adipose tissue from obese subjects.

Interest in adipose tissue pathophysiology and biochemistry have expanded considerably in the past two decades due to the ever increasing and alarming rates of global obesity and its critical outcome defined as metabolic syndrome (MS). This obesity-linked systemic dysfunction generates high risk factors of developing perilous diseases like type 2 diabetes, cardiovascular disease or cancer. Amino acids could play a crucial role in the pathophysiology of the MS onset. Focus of this study was to fully characterize amino acids metabolome modulations in visceral adipose tissues (VAT) from three adult cohorts: (i) obese patients (BMI 43-48) with metabolic syndrome (PO), (ii) obese subjects metabolically well (O), and (iii) non obese individuals (H). 128 metabolites identified as 20 protein amino acids, 85 related compounds and 13 dipeptides were measured by ultrahigh performance liquid chromatography-tandem mass spectroscopy (UPLC-MS/MS) and gas chromatography-/mass spectrometry GC/MS, in visceral fat samples from a total of 53 patients. Our analysis indicates a probable enhanced BCAA (leucine, isoleucine, valine) degradation in both VAT from O and PO subjects, while levels of their oxidation products are increased. Also PO and O VAT samples were characterized by: elevated levels of kynurenine, a catabolic product of tryptophan and precursor of diabetogenic substances, a significant increase of cysteine sulfinic acid levels, a decrease of 1-methylhistidine, and an up regulating trend of 3-methylhistidine levels. We hope this profiling can aid in novel clinical strategies development against the progression from obesity to metabolic syndrome.

Energy drink consumption: a survey in high school students and associated psychological effects.

INTRODUCTION: Energy drinks represent an emerging health problem among young people. Energy drinks generally refer to a class of beverages containing sugars and various combinations of bioactive ingredients such as caffeine, taurine etc. Also the mix of energy drinks with alcohol is fairly frequent among young people and could be associated with dangerous effects. METHODS: In 2016-2017, a cross-sectional study was conducted in 1581 students attending eight high school in the Marche Region. Data were collected via an anonymous self-administered questionnaire. RESULTS: The 27.7% of students use energy drinks and the majority, corresponding with the 93.0%, are aware of the main ingredients contained in energy drinks. The main activities for which young people use these drinks are: sport, leisure, pleasure, study. Young people who admit to using alcohol mixed with energy drinks more than 4 times a month are an alarming fact. CONCLUSIONS: This research confirms that energy drinks are used more by young males and especially by those who practice sports. Furthermore, the use these beverages to increase the concentration in the study and to be more brilliant in free time, is confirmed.

Implications of modified food choices and food-related lifestyles following the economic crisis in the Marche Region of Italy.

BACKGROUND: The economic crisis in Italy has led to profound changes in resource management not only at the macro level but also for individual families, causing substantial changes in different habits of Italians. STUDY DESIGN: The purpose of this research was to conduct a study on changes in family eating habits potentially triggered by the economic crisis was conducted in an area of the Marche Region in central Italy. METHODS: The research was conducted in the period 2016 - 2017 by administering a specific and anonymous questionnaire. RESULTS: The interviewed people has reduced its food consumption. In particular, analyzing the results for the animal protein food group, there has been a reduction in purchase of beef, and an increase in that of pork. Overall fish consumption has decreased by 44%, with a decrease in the purchase of fresh fish, and an increase in that of canned fish products. Finally, consumers have reduced their purchases of fresh and canned legumes, fresh vegetables, and fresh fruit. CONCLUSION: The economic crisis seems to have changed the eating habits and food-related lifestyle choices of the subjects studied, especially in the urban area affected by the deeper economic depression. These changes are likely to have permanent social consequences, and deserve to be analyzed also in smaller territories in order to better understand the dynamics of individual choices and the social framework.

Presence of Legionella spp. in human dental plaque.

AIMS: The aim of this research is to verify the presence of Legionella in human dental plaque. METHODS: 65 adult patients not treated with systemic or local antibiotics at least 2 months before the time of sample collection were enrolled for plaque collection between September 2015 and December 2016. A brief questionnaire about lifestyle and health risks was administered. Legionella spp. detection has been executed by semi- nested PCR. RESULTS: 8 out of 65 plaque samples (12.3%) were positive for Legionella spp. As regards health risks and lifestyle aspects, no relevant difference was observed between patients involved in our study, except for two positive patients who have reported a COPD ongoing and a pneumonia in the past. CONCLUSIONS: This study represents a step forward in the knowledge of reservoirs of the microorganism and richness of oral microbiota.

Arterial ageing: from endothelial dysfunction to vascular calcification.

Complex structural and functional changes occur in the arterial system with advancing age. The aged artery is characterized by changes in microRNA expression patterns, autophagy, smooth muscle cell migration and proliferation, and arterial calcification with progressively increased mechanical vessel rigidity and stiffness. With age the vascular smooth muscle cells modify their phenotype from contractile to 'synthetic' determining the development of intimal thickening as early as the second decade of life as an adaptive response to forces acting on the arterial wall. The increased permeability observed in intimal thickening could represent the substrate on which low-level atherosclerotic stimuli can promote the development of advanced atherosclerotic lesions. In elderly patients the atherosclerotic plaques tend to be larger with increased vascular stenosis. In these plaques there is a progressive accumulation of both lipids and collagen and a decrease of inflammation. Similarly the plaques from elderly patients show more calcification as compared with those from younger patients. The coronary artery calcium score is a well-established marker of adverse cardiovascular outcomes. The presence of diffuse calcification in a severely stenotic segment probably induces changes in mechanical properties and shear stress of the arterial wall favouring the rupture of a vulnerable lesion in a less stenotic adjacent segment. Oxidative stress and inflammation appear to be the two primary pathological mechanisms of ageing-related endothelial dysfunction even in the absence of clinical disease. Arterial ageing is no longer considered an inexorable process. Only a better understanding of the link between ageing and vascular dysfunction can lead to significant advances in both preventative and therapeutic treatments with the aim that in the future vascular ageing may be halted or even reversed.

The possible role of glutathione-S-transferase activity in diabetic nephropathy.

The most common cause of end stage renal disease is diabetic nephropathy. An early diagnosis may allow an intervention to slow down disease progression. Recently, it has been hypothesized that glutathione-S-transferase (GST) activity may be a marker of severity of chronic kidney disease. In particular, a lower GST activity is present in healthy subjects compared to patients with nephropathy. In the present review we illustrate the scientific evidence underlying the possible role of GST activity in the development of diabetic nephropathy and we analyze its usefulness as a possible early biomarker of this diabetic complication.

Gut hormones and endothelial dysfunction in patients with obesity and diabetes.

Overweight and obesity are the fifth leading risk for global deaths and its prevalence has doubled since 1980. At least 2.8 million adults, worldwide, die each year as a result of being overweight or obese. The deleterious effects of obesity are tightly related to diabetes, as they are often clinically present in combination to confer increased cardiovascular mortality. Thus, patients with diabetes and obesity are known to develop accelerated atherosclerosis characterized by a dysfunctional endothelium and decreased nitric oxide bioavailability. Recent clinical studies support, indeed, the use of incretin-based antidiabetic therapies for vascular protection. Thus, attention has been focusing on gut hormones and their role, not only in the regulation of appetite but also in vascular health. Intervention directed at modulating these molecules has the potential to decrease mortality of patients with diabetes and obesity. This review will cover part of the ongoing research to understand the role of gut hormones on endothelial function and vascular health.

Obesity, inflammation and endothelial dysfunction.

Cardiovascular disease is the leading cause of morbidity and mortality in obese individuals. Obesity dramatically increases the risk of development of metabolic and cardiovascular disease. This risk appears to originate from disruption in adipose tissue function leading to a chronic inflammatory state and to dysregulation of the endocrine and paracrine actions of adipocyte-derived factors. These, in turn, impair vascular homeostasis and lead to endothelial dysfunction. An altered endothelial cell phenotype and endothelial dysfunction are common among all obesity-related complications. A crucial aspect of endothelial dysfunction is reduced nitric oxide (NO) bioavailability. A systemic pro-inflammatory state in combination with hyperglycemia, insulin resistance, oxidative stress and activation of the renin angiotensin system are systemic disturbances in obese individuals that contribute independently and synergistically to decreasing NO bioavailability. On the other hand, pro-inflammatory cytokines are locally produced by perivascular fat and act through a paracrine mechanism to independently contribute to endothelial dysfunction and smooth muscle cell dysfunction and to the pathogenesis of vascular disease in obese individuals. The promising discovery that obesity-induced vascular dysfunction is, at least in part, reversible, with weight loss strategies and drugs that promote vascular health, has not been sufficiently proved to prevent the cardiovascular complication of obesity on a large scale. In this review we discuss the pathophysiological mechanisms underlying inflammation and vascular damage in obese patients.

Novel therapeutic approaches based on the pathological role of gut dysbiosis on the link between nonalcoholic fatty liver disease and insulin resistance.

The growing global epidemic of obesity and type 2 diabetes mellitus has determined an increased prevalence of NAFLD (non-alcoholic fatty liver disease), making it the most common chronic liver disease in the Western world and a leading cause of liver transplantation. In the last few years, a rising number of studies conducted both on animal and human models have shown the existence of a close association between insulin resistance (IR), dysbiosis, and steatosis. However, all the mechanisms that lead to impaired permeability, inflammation, and fibrosis have not been fully clarified. Recently, new possible treatment modalities have received much attention. To reach the review purpose, a broad-ranging literature search on multidisciplinary research databases was performed using the following terms alone or in combination: "NAFLD", "gut dysbiosis", "insulin resistance", "inflammation", "probiotics", "Chinese herbs". The use of probiotics, prebiotics, symbiotics, postbiotics, fecal microbiota transplant (FMT), Chinese herbal medicine, antibiotics, diet (polyphenols and fasting diets), and minor therapies such as carbon nanoparticles, the MCJ protein, water rich in molecular hydrogen, seems to be able to improve the phenotypic pattern in NAFLD patients. In this review, we provide an overview of how IR and dysbiosis contribute to the development and progression of NAFLD, as well as the therapeutic strategies currently in use.

The possible role of physical activity in the modulation of gut microbiota in chronic kidney disease and its impact on cardiovascular risk: a narrative review.

Chronic degenerative non-communicable diseases (CDNCDs), in particular chronic kidney disease, induce gut microbiota (GM) dysbiosis, which, in turn, worsens the progression of CDNCDs and patients' quality of life. We analyzed literature studies to discuss the possible positive and beneficial impact of physical activity on GM composition and CV risk in CKD patients. Regular physical activity seems to be able to positively modulate the GM, reducing the systemic inflammation and consequently the production of uremic gut-derived toxins, which are directly correlated with the increase of cardiovascular risk. In particular, the accumulation of indoxyl sulphate (IS) seems to be able to induce vascular calcifications, vascular stiffness and cardiac calcifications, while p-Cresyl sulphate (p-CS) seems to be able to exert a cardiotoxic action through metabolic pathways, capable of inducing oxidative stress. In addition, trimethylamine N-oxide (TMAO) can alter lipid metabolism, inducing the production of foam cells and causing an accelerated atherosclerosis process. In this context, a regular physical activity program seems to represent an adjuvant non-pharmacological approach to the clinical management of CKD patients.

Screening of Fabry Disease of patients in renal replacement therapy in a population from Lazio (Italy).

OBJECTIVE: Fabry's disease (FD) is a genetic disorder of lysosomal storage characterized by the intralysosomal accumulation of globotriaosylceramide (Gb3). This genetic mutation causes a total or partial deficit of the α-galactosidase (GAL) enzyme activity. FD has an incidence of 1:40000-60000 born alive. Its prevalence is higher in specific pathological conditions like chronic kidney disease (CKD). The aim of this study was to evaluate the FD prevalence in Italian renal replacement therapy (RRT) patients from Lazio region. PATIENTS AND METHODS: 485 patients in RRT (hemodialysis, peritoneal dialysis, and kidney transplantation) were recruited. The screening test was performed on venous blood sample. The latter was analyzed using specific FD diagnostic kit, based on the analysis of dried blood spots on filter paper. RESULTS: We found 3 cases of positivity to FD (1 female and 2 males). In addition, 1 male patient was identified with biochemical alteration indicative of GAL enzyme deficiency with a genetic variant of the GLA gene of unknown clinical significance. The FD prevalence in our population was 0.60% (1 case out 163), it rises to 0.80% (1 case out of 122) if the genetic variant of unknown clinical significance is considered. Comparing the three subpopulations, we observed a statistically significant difference in GAL activity in transplanted patients compared to dialysis patients (p<0.001). CONCLUSIONS: Considering the presence of an enzyme replacement therapy able to modify FD clinical history, it is essential to try to implement FD early diagnoses. However, the screening is too expensive to be extended on large scale, due to the low prevalence of the pathology. The screening should be performed on high-risk populations.

Insulinemia and the risk of breast cancer and its relapse.

A fairly large body of evidence has shown that insulin is a tumour-promoting agent, especially for breast cancer. High circulating and microenvironmental levels of insulin may directly increase the risk of breast cancer via the activation of cognate receptors expressed on normal and cancer cells and indirectly be associated with other known metabolic risk factors of cancer that usually are present in conjunction with the hyperinsulinic state. The focus of this review is to analyse and discuss available data in the literature on the possible causative/prognostic role of insulin resistance/hyperinsulinemia in breast cancer development and progression.

Environmental risk factors for amyotrophic lateral sclerosis: methodological issues in epidemiologic studies.

The exact role of environmental risk factors in the etiology of the neurodegenerative disease amyotrophic lateral sclerosis (ALS) is still unknown. Their hypothetical contribution ranges from a minimal impact to a major role. Among the environmental factors strictu sensu (i.e., not life-style factors) suspected to play a role in ALS etiology, we consider pesticides, the metalloid selenium, some heavy metals, magnetic fields and cyanobacteria. However, the possibility exists that these factors exert their activity only in genetically susceptible persons and only after long-term exposures, thus further hampering epidemiologic studies. The recent availability of powerful tools such as population-based ALS registries for case ascertainment and clustering detection, and of environmental modeling techniques and of geographical information systems, may yield unique opportunities for offering insight into the etiology of the disease.

Dental unit water content and antibiotic resistance of Pseudomonas aeruginosa and Pseudomonas species: a case study.

Background: Many studies consider the contamination of dental unit waterlines (DUWLs), but few of them have studied the possible presence of antibiotic resistant Pseudomonas aeruginosa in the DUWLs. Aims: Investigation of the presence of P. aeruginosa and Pseudomonas spp. strains in DUWLs and evaluation of their resistance to six antibiotics (ceftazidime, netilmicin, piperacillin/tazobactam, meropenem, levofloxacin, colistin sulfate) at a public dental clinic in Milan, Italy. Results: Dental units were contaminated by P. aeruginosa with loads of 2-1,000 CFU/L and were mainly located on the mezzanine floor, with a range of 46-54%, while Pseudomonas spp. were primarily found on the first and second floors, ranging from 50 to 91%. P. aeruginosa was antibiotic resistant in 30% of the strains tested, andPseudomonas spp. in 31.8% . Cold water from controls was also contaminated by these microorganisms. Conclusion: Monitoring antibiotic resistance in the water and adopting disinfection procedures on DUs are suggested within the Water Safety Plan.

Verapamil: prevention and treatment of cardio-renal syndromes in diabetic hypertensive patients?

Patients with diabetes mellitus (DM) often present other chronic comorbidities including arterial hypertension (AH), chronic kidney disease (CKD), ischemic heart disease (IHD) and heart failure with preserved ejection fraction (HFpEF). The frequent association of the latter conditions is considered part of the spectrum of cardio-renal syndromes (CRS), a group of disorders of the heart and kidneys whereby acute or chronic dysfunction in one organ may induce acute or chronic dysfunction of the other. Verapamil is a non-dihydropyridine calcium channel blocker (CCB) widely used in the treatment of hypertension, chronic stable angina, secondary prevention of reinfarction, paroxysmal supra-ventricular tachycardia and for rate control in atrial fibrillation/flutter. In addition to its antihypertensive and anti-ischemic actions verapamil exerts favorable effects also on glycemic control, proteinuric diabetic nephropathy, left ventricular diastolic dysfunction and sympathetic nervous system overactivity which may potentially benefit patients with DM and CRS. In this narrative review, we summarize the current evidence on the potential role of verapamil in the prevention and treatment of CRS in diabetic hypertensive patients.

[Hygienic profile of the water in Milan swimming pools: a three-year comparative study]. / Profilo igienico-sanitario dell'acqua delle piscine milanesi: tre anni a confronto.

The aim of this work was to determine whether swimming pool water quality in Milan from 2006 to 2008 was within the standards established by national and local Italian laws (Circolare Min. Sanità 128/71 and DGR 2552/2006). In 2006, 580 samples of water from public swimming pools were analyzed to determine the presence of heterotrophic counts at 37 degrees and total coliforms; pH, free chlorine and chloride of each sample were also measured. In the following years, water from both public and private swimming pools were examined to measure heterotrophic count at 22 degrees and 36 degrees, Escherichia coli, enterococci, Pseudomonas aeruginosa, Staphylococcus aureus, pH, free chlorine, and nitrates. The total number of analyses carried out in 2007 and 2008 was 2074 and 1532, respectively. In 2006, the extent of noncompliance of all swimming pools that was observed for both physical/chemical and microbiological parameters was 72.3%, which then decreased to 53.2% and 36.2% in 2007 and 2008, respectively. In particular with regard to the microbiological analysis, an increase of noncompliance based on at least one parameter was determined (7.1% in 2006 vs. 21.5% in 2007 and 22% in 2008). In contrast, a decrease of the extent of noncompliance based on at least one physical/chemical parameter was observed (from 68.1% in 2006 to 40.4% and 22.3% in 2007 and 2008, respectively). Interestingly, public swimming pools exceeded the legal limits of microbiological concentration more often than the private ones, whereas both types of swimming pools showed a decrease in noncompliance with regard to the physical/chemical parameters.

"The Linosa Study": epidemiological and heritability data of the metabolic syndrome in a Caucasian genetic isolate.

BACKGROUND AND AIMS: Growing evidence suggests that the metabolic syndrome (MetS) has both a genetic and environmental basis. To evaluate the possibility of a further genetic analysis, we estimated prevalence rates and heritabilities for the MetS and its individual traits in the adult population of Linosa, a small and isolated Italian Island in the southern-central part of the Mediterranean Sea. METHODS AND RESULTS: The Linosa Study (LiS) group consisted of 293 Caucasian native subjects from 51 families (123 parents; 170 offsprings). The MetS was defined according to NCEP/ATP III criteria and the following prevalence rates were calculated: hyperglycaemia 20.3%; central obesity 34.9%; hypertension 43.4%; hypertriglyceridaemia 29.9%; "low HDL" 56.6%; MetS 29.9%. Waist circumference was significantly related to all the quantitative parameters included in the NCEP/ATP III MetS definition. The MetS showed a heritability of 27% (p=0.0012) and among its individual components, treated as continuous and discrete traits, heritability ranged from 10% for blood glucose to 54% for HDL-cholesterol. Among MetS subtypes, the clustering of central obesity, hypertriglyceridaemia and "Iow HDL" had the highest heritability (31%; p<0.001). CONCLUSION: These data showed high prevalence rates for the MetS and its related traits in an isolated and small Caucasian population. The appreciable heritability estimates for the MetS and some of its components/clusters in the LiS population might support the observation of genetic factors underlying the pathogenesis of the MetS and encourage further analysis to identify new susceptibility genes.

[Endothelial dysfunction and diabetes: possible role in kidney damage]. / Danno endoteliale e malattia diabetica: possibile ruolo nella nefropatia.

Endothelial damage, with loss of the vascular protective effects of nitric oxide (NO), is an important early step in the development of microvascular and macrovascular complications of diabetes. Endothelial dysfunction is closely associated with diabetic nephropathy in type 1 and 2 diabetes. In this review we will discuss the mechanisms by which hyperglycemia may cause kidney damage and endothelial dysfunction. Hyperglycemia causes microvascular dysfunction, which contributes to the development of end stage renal disease. Determining the role of endothelial abnormalities in the development of diabetic nephropathy is critical to understanding the etiology and pathogenesis of the microvascular complications of diabetes. Endothelial function can be assessed by invasive and noninvasive techniques both in the coronary and peripheral circulation. Endothelial dysfunction is considered a reversible phenomenon; pharmacological intervention with hypolipidemic agents, insulin sensitizers, ACE inhibitors and angiotensin II receptor blockers (ARB) as well as dietary and lifestyle modifications have been shown to reverse it.

[Diabetes worsens the clinical manifestations and prognosis of concurrent cardiovascular and kidney disease]. / Come il diabete peggiora espressioni e prognosi della malattia cardio-vasculo-renale.

Numerous studies have shown a marked increase in the incidence of diabetes mellitus worldwide. Diabetes mellitus is currently considered equivalent to coronary artery disease in terms of prognostic risk stratification, and its high prevalence makes this clinical condition the first cause of end-stage renal disease requiring chronic hemodialysis or kidney transplant. Even if chronic kidney disease remains the ''Cinderella of the cardiovascular profile'', the presence of microalbuminuria is closely related to a high risk of development of coronary artery disease. The same risk factors that impair heart function are also harmful to the kidney, and the common pathophysiological features of the two systems are at the origin of a new subspecialty, cardionephrology. A crucial task of cardiologists and nephrologists is the early identification of high risk patients with concurrent cardiovascular and kidney disease. The utilization of simple screening methods such as assessment of microalbuminuria and glomerular filtration rate by family doctors may help in establishing prevention strategies directed towards cardiovascular risk and progression of kidney disease. In conclusion, early stratification of cardiovascular risk, coupled with primary prevention strategies aimed at the general population, is warranted to obtain a significant reduction of kidney and cardiovascular disease and of the need for chronic hemodialysis treatment. This strategy is safe and cost-effective in comparison with the costs of chronic dialysis of patients affected by chronic kidney disease.

[Investigation on strains of Legionella pneumophila, isolated from a hospital of Milano, with three genotyping methods]. / Genotipizzazione di ceppi di Legionella pneumophila, isolati in strutture sanitarie di differenti tipologie.

Various techniques have been developed in recent years for the molecular typing of microorganisms. Remains particularly difficult to isolate clinical strains for the low availability of cases and even more problematic matching clinical / environmental strains. We investigated 13 strains of Legionella pneumophila of clinical and environmental origin, isolated in 3 Health Facilities in Milan (2003-2006), using three molecular typing methods: Pulse-Field Gel Electrophoresis, Amplified Fragment Length Polymorphism and Sequence-Based Typing. PFGE and AFLP showed the correlation between a clinical case with only one of the environmental isolates taken from the places frequented by the patient, demonstrating with certainty the nosocomial origin of the case and identifying the source of infection in the shower water (Clin. 1 and Env. 1N). Two clinical samples from patients admitted to different wards presented an identical profile, which suggests that the nosocomial origin assumed an epidemic form, even without having isolated the environmental strain due to the absence of samples drawn during the period under consideration (Clin. 2 and 3). Finally, the comparison between the isolated environmental strains demonstrated a heterogeneous presence of strains, not correlated to each other although they belong to the same serum-group, having profiles that are clearly different regarding number and position of bands (Env. 2 and 4). The profile 2,10,18,10,1,1 had never been isolated and typed previously in Europe. The SBT has proved a better technique for reproducibility and interpretation of results than PFGE and AFLP To complete studies on SBT method, now considered gold standard, is currently being the EWGLI 5th Proficiency Panel, in which we are actively involved with the genotyping of five strains according to the latest version of the protocol (4.1).