Chronic exposure to low dose of bisphenol a impact on the firts round of spermatogenesis via sirt1 modulation

Spermatogenesis depends on endocrine, autocrine and paracrine communications along the hypothalamus-pituitary-gonad axis. Bisphenol A (BPA), an estrogen-mimic endocrine disrupting chemical, is an environmental contaminant used to manufacture polycarbonate plastics and epoxy resins with toxic effects for male reproduction. Here we investigated whether the chronic exposure to low BPA doses affects spermatogenesis through the modulation of SIRT1, a NAD +-dependent deacetylase involved in the progression of spermatogenesis, with outcomes on apoptosis, oxidative stress, metabolism and energy homeostasis. BPA exposure via placenta first, and lactation and drinking water later, affected the body weight gain in male offspring at 45 postnatal days and the first round of spermatogenesis, with impairment of blood testis barrier, reactive oxygen species production, DNA damage and decreased expression of SIRT1. The analysis of SIRT1 downstream molecular pathways revealed the increase of acetyl-p53 Lys370 , γH2AX foci, the decrease of oxidative stress defenses and the higher apoptotic rate in the testis of treated animals, with partial rescue at sex maturation. In conclusion, SIRT1 pathways disruption after BPA exposure can have serious consequences on the first round of spermatogenesis. Spermatogenesis is a complex biological process that requires the self-renewal and the commitment of dip-loid spermatogonia for proliferation, the meiosis of spermatocytes and the post-meiotic differentiation of hap-loid spermatids into mature spermatozoa. This process strongly depends on endocrine control through the hypothalamus-pituitary-gonad (HPG) axis and requires intratesticular autocrine and paracrine communications between germ and somatic cells 1-5. Environment and diet influence spermatogenesis with known consequences on male fertility 6. In this respect, a variety of natural and synthetic compounds, collectively named endocrine disrupting chemicals (EDCs), mimics or antagonizes endogenous hormones, interferes with hormone synthesis and clearance, and generates adverse health outcomes in mammals 7-9. Bisphenol A [2,2-bis (4-hydroxyphenyl) propane, BPA], an estrogen-mimic EDC, is an ubiquitous environmental contaminant used as monomer to manufacture polycarbonate plastics and epoxy resins. It is commonly used to produce plastics that line food/drink containers, goods of common use, thermal receipts, and medical devices such as dental sealants. Heat (as in microwaves or dishwashers) and either acidic or basic conditions (as in foods) accelerate the hydrolysis of ester bonds linking BPA monomers leading to the release of BPA in the environment with potential risk of exposure for living beings. BPA is a well-known toxicant for male reproductive physiology in animal models, but data in humans are quite controversial since exposure doses, duration, route and life stage all affect BPA effects

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Comparative assessment of the quality of commercial black and green tea using microbiology analyses

Background: Drinking tea constitutes a tradition which is deeply rooted in the culture of several countries. Moreover, in recent years, tea consumption is growing all over the world. Improper herbal tea storage (long periods, humid environments) represents a relevant health hazard for consumers because of the growth of bacteria and molds. Results: This study analyzed 32 samples of commercially available black and green teas - purchased from southern Italy markets and online-shops - and the monitoring of microbiological quality of the tea bag content was performed. Evaluations were conducted with the aim of characterizing pathogens indicated by the European and American guidelines (total bacterial count, fungi and Escherichia coli) and on the research of Pseudomonas spp. and Clostridium perfringens. The presence of ochratoxin A in tea matrix-leaves and infusions was further assessed, using a validated and accredited HPLC-FLD method. Microbial loads, for over 80% samples, ranged from 1.0 × 102 to 2.8 × 105 CFU/g tea: most of identified microorganisms were classified as Bacillaceae. The utilization of rapid detection and identification methods (PCR and sequencing), allowed the characterization of strains of Pseudomonas psychrotolerans, Staphylococcus warneri, Pantoea gaviniae and the isolation of one strain of Clostridium perfringens, whose ability to produce toxins can result in harmful outcomes for consumers. Fungi were isolated from 70% samples: the most prevalent molds were Aspergillus niger strains, followed by Aspergillus tubingensis. Ochratoxin A was detected in 22 of 32 tea solid samples investigated: concentrations resulted over the indicated limits for food products for 50% samples. Conclusions: Results obtained demonstrated the need to develop targeted regulations for the safety of herbal teas.

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Metabolomic Signature of Endometrial Cancer

Endometrial cancer (EC) is the most common cancer of the female reproductive tract in developed countries. At the moment, no effective screening system is available. Here, we evaluate the diagnostic performance of a serum metabolomic signature. Two enrollments were carried out, one consisting of 168 subjects: 88 with EC and 80 healthy women, was used for building the classification models. The second (used to establish the performance of the classification algorithm) was consisted of 120 subjects: 30 with EC, 30 with ovarian cancer, 10 with benign endometrial disease, and 50 healthy controls. Two ensemble models were built, one with all EC versus controls (Model I) and one in which EC patients were aggregated according to their histotype (Model II). Serum metabolomic analysis was conducted via gas chromatography–mass spectrometry, while classification was done by an ensemble learning machine. Accuracy ranged from 62% to 99% for the Model I and from 67% to 100% for the Model II. Ensemble model showed an accuracy of 100% both for Model I and II. The most important metabolites in class separation were lactic acid, progesterone, homocysteine, 3-hydroxybutyrate, linoleic acid, stearic acid, myristic acid, threonine, and valine. The serum metabolomics signature of endometrial cancer patients is peculiar because it differs from that of healthy controls and from that of benign endometrial disease and from other gynecological cancers (such as ovarian cancer).

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hsp70 AS New Cadmium Bioaccumulation Marker to prevent the risks of Mussels Consumption in Human Nutrition


The consumption of seafood has increased in recent years, especially in coastal regions. The consumption of mussels provides proteins, essential minerals and vitamins, and thus, some protection from certain diseases but the risks and benefits of their consumption are still hard to assess because of the metals bioaccumulated from the marine environment, with their toxicity. Mussels accumulate a wide range of metals, included cadmium, in their soft tissue. Cadmium is a heavy metal particularly hazardous for human health and is an important pollutant in estuarine and coastal environments. Thus, the determination of the concentrations of cadmium in mussels is essential because of their usage as seafood and the potential adverse effects of their consumption on human health. In order to identify a quick cadmium bioaccumulation marker usable in monitoring programs, we analyzed the metal content in Mytilus galloprovincialis gill tissues and its relationship with hsp70 expression levels after a laboratory exposure for 24 h to 1,5; 5 and 10 μM CdCl2 in artificial sea water. Inductively coupled plasma-mass spectrometry showed that cadmium content increased in gills tissues in an exposure dose-dependent fashion. RT-qPCR, showed that Cd exposure induced hsp70 increase resulting in 3,1;10 and 12 fold at 1,5; 5, 10 μM, respectively in comparison with unexposed mussels. Finally, hsp70 expression levels correlated with the amount of bioaccumulated cadmium in gill tissue, indicating hsp70 as a potential marker, even if not univocally, of significative cadmium bioaccumulation usable in environmental monitoring programs and for seafood safety.

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Nutrient, fatty acid and mineral composition of selected white food-grade sorghum hybrids grown in a Mediterranean area of Southern Italy

The nutrient composition, fatty acid content and mineral content of six white food-grade sorghum hybrids selected in either Argentina or Bolivia, and grown in a Mediterranean area of Southern Italy were studied. The six hybrids were analyzed for various attributes including moisture, protein, carbohydrate, dietary fiber, fat contents, fatty acid composition, and mineral content. Slight variations in both protein and in fiber contents were observed among hybrids. Linoleic, oleic and palmitic were the most abundant fatty acids in all samples with noticeable difference in their percentage content between both hybrids from Argentina and Bolivia. Slight variations in the content of the elements were found among the six white sorghum hybrids examined, and K, Fe and Sb were, respectively, the most abundant macro-element, micro-element and trace element in all analyzed hybrids. These results are discussed in the context of the importance of the white food-grade sorghum in human nutrition, and of the opportunity to select the best varieties from the point of view of nutritional content.

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A Metabolomics-based approach for non-invasive diagnosis of chromosomal anomalies

Introduction: Chromosomal anomalies (CA) are the most frequent fetal anomalies.
Objective: To evaluate the diagnostic performance of a machine learning ensemble model based on the maternal serum metabolomic fingerprint of fetal aneuploidies during the second trimester .
Methods: This is a case-control pilot study. Metabolomic profiles have been obtained on serum of 328 mothers (220 controls and 108 cases), using gas chromatography coupled to mass spectrometry. Eight machines learning and classification models were built and optimized. An ensemble model was built using a voting scheme. All samples were randomly divided into two sets. One was used as training set, the other one for diagnostic performance assessment.
Results: Ensemble machine learning model correctly classified all cases and controls. The accuracy was the same for trisomy 21 and 18; also, the other CA were correctly detected. Elaidic, stearic, linolenic, myristic, benzoic, citric and glyceric acid, mannose, 2-hydroxy butyrate, phenylalanine, proline, alanine and 3-methyl histidine were selected as the most relevant metabolites in class separation.
Conclusion: The proposed model, based on the maternal serum metabolomic fingerprint of fetal aneuploidies during the second trimester, correctly identifies all the cases of chromosomal abnormalities. Overall, this preliminary analysis appeared suggestive of a metabolic environment conductive to increased oxidative stress and a disturbance in the fetal central nervous system development. Maternal serum metabolomics can be a promising tool in the screening of chromosomal defects. Moreover, metabolomics allows to extend our knowledge about biochemical alterations caused by aneuploidies and responsible for the observed phenotypes.

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Bisphenol A in reproduction: epigenetic effects

Background: Bisphenol A (BPA) is an endocrine disrupting chemical widely used in the manufacture of polycarbonate plastic and epoxy resin to produce a multitude of consumer products, food and drink containers, and medical devices. BPA is similar to estradiol in structure and thus interferes in steroid signalling with different outcomes on reproductive health depending on doses, life stage, mode, and timing of exposure. In this respect, it has an emerging and controversial role as a "reproductive toxicant" capable of inducing short and long-term effects including the modulation of gene expression through epigenetic modification (i.e. methylation of CpG islands, histone modifications and production of non-coding RNA) with direct and trans-generational effects on exposed organisms and their offspring, respectively. Objective: This review provides an overview about BPA effects on reproductive health and aims to summarize the epigenetic effects of BPA in male and female reproduction. Results: BPA exerts epigenetic effects in both male and female reproduction. In males, BPA affects spermatogenesis and sperm quality and possible trans-generational effects on the reproductive ability of the offspring. In females, BPA affects ovary, embryo development, and gamete quality for successful in vivo and in vitro fertilization (IVF). Conclusion: The exact mechanisms of BPA - mediated effects in reproduction are not fully understood; however, the environmental exposure to BPA - especially in fetal and neonatal period - deserves attention to preserve the reproductive ability in both sexes and to reduce the epigenetic risk for the offspring.

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What is in your cup of tea? DNA Verity Test to characterize black and green commercial teas

In this study, we used several molecular techniques to develop a fast and reliable protocol (DNA Verity Test, DVT) for the characterization and confirmation of the species or taxa present in herbal infusions. As a model plant for this protocol, Camellia sinensis, a traditional tea plant, was selected due to the following reasons: its historical popularity as a (healthy) beverage, its high selling value, the importation of barely recognizable raw product (i.e., crushed), and the scarcity of studies concerning adulterants or contamination. The DNA Verity Test includes both the sequencing of DNA barcoding markers and genotyping of labeled-PCR DNA barcoding fragments for each sample analyzed. This protocol (DVT) was successively applied to verify the authenticity of 32 commercial teas (simple or admixture), and the main results can be summarized as follows: (1) the DVT protocol is suitable to detect adulteration in tea matrices (contaminations or absence of certified ingredients), and the method can be exported for the study of other similar systems; (2) based on the BLAST analysis of the sequences of rbcL+matK±rps7-trnV (GAC) chloroplast markers, C. sinensis can be taxonomically characterized; (3) rps7-trnV (GAC) can be employed to discriminate C. sinensis from C. pubicosta; (4) ITS2 is not an ideal DNA barcode for tea samples, reflecting potential incomplete lineage sorting and hybridization/introgression phenomena in C. sinensis taxa; (5) the genotyping approach is an easy, inexpensive and rapid pre-screening method to detect anomalies in the tea templates using the trnH (GUG)-psbA barcoding marker; (6) two herbal companies provided no authentic products with a contaminant or without some of the listed ingredients; and (7) the leaf matrices present in some teabags could be constituted using an admixture of different C. sinensis haplotypes and/or allied species (C. pubicosta).

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Urinary metabolomics in pediatric obesity and NAFLD identifies metabolic pathways/metabolites related to dietary habis and gut-liver axis perturbations

To get insight into still elusive pathomechanisms of pediatric obesity and non-alcoholic fatty liver disease (NAFLD) we explored the interplay among GC-MS studied urinary metabolomic signature, gut liver axis (GLA) abnormalities, and food preferences (Kid-Med). Intestinal permeability (IP), small intestinal bacterial overgrowth (SIBO), and homeostatic model assessment-insulin resistance were investigated in forty children (mean age 9.8 years) categorized as normal weight (NW) or obese (body mass index <85th or >95th percentile, respectively) ± ultrasonographic bright liver and hypertransaminasemia (NAFLD). SIBO was increased in all obese children (p = 0.0022), IP preferentially in those with NAFLD (p = 0.0002). The partial least-square discriminant analysis of urinary metabolome correctly allocated children based on their obesity, NAFLD, visceral fat, pathological IP and SIBO. Compared to NW, obese children had (1) higher levels of glucose/1-methylhistidine, the latter more markedly in NAFLD patients; and (2) lower levels of xylitol, phenyl acetic acid and hydroquinone, the latter especially in children without NAFLD. The metabolic pathways of BCAA and/or their metabolites correlated with excess of visceral fat centimeters (leucine/oxo-valerate), and more deranged IP and SIBO (valine metabolites). Urinary metabolome analysis contributes to define a metabolic fingerprint of pediatric obesity and related NAFLD, by identifying metabolic pathways/metabolites reflecting typical obesity dietary habits and GLA perturbations

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Multiple gut-liver axis abnormalities in children with obesity with and without hepatic involvement

BACKGROUND: Gut-liver axis (GLA) dysfunction appears to play a role in obesity and obesity-related hepatic complications.

OBJECTIVES: This study sought to concurrently explore several GLA components in a paediatric obese population with/without liver disease.

METHODS: Thirty-two children (mean age 11.2 years) were enrolled: nine controls with normal weight and 23 patients with obesity (OB+). Of the 23 patients OB(+), 12 had not steatosis (ST-), and 11 had steatosis (ST+) (associated [n = 8] or not [n = 3] with hypertransaminasaemia [ALT +/-]). Subjects were characterized by using auxologic, ultrasonographic and laboratory parameters. A glucose hydrogen breath test was performed to test for small intestinal bacterial overgrowth, a urinary lactulose/mannitol ratio (LMR) was obtained to assess intestinal permeability, and tests for transaminases, blood endogenous ethanol, endotoxin and faecal calprotectin were also conducted.

RESULTS: Eleven out of 23 patients OB(+) (p < 0.05) exhibited pathological (>90th percentile of the control group values) LMR, with values paralleling the grade of liver involvement (normal weight < OB[+] < OB[+]ST[+]ALT[-] < OB[+)]ST[+]ALT[+] [p < 0.05]). LMR significantly correlated with ethanolaemia (r = 0.38, p = 0.05) and endotoxaemia (r = 0.48, p = 0.015) concentrations. Increased permeability was a risk factor for the development of steatosis (p < 0.002). SIBO was present only in patients with obesity. Faecal calprotectin concentrations were within normal limits in all subjects.

CONCLUSIONS:Increased permeability, endogenous ethanol and systemic endotoxin concentrations reflect some GLA dysfunction in obesity and its hepatic complications. Pending further results to establish their potential causative roles, the modulation of the GLA appears to represent a possible target for the prevention and treatment of these conditions.

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Simultaneous Doppler ultrasound fetal cerebrovascular and maternal glucose homeostasis evaluation during normal pregnancy.

Simultaneous Doppler ultrasound fetal cerebrovascular and maternal glucose homeostasis evaluation during normal pregnancy.


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Evaluation of a mathematical model for glycemia prediction as a marker of intravasation during hysteroscopy

Background: Intravasation is one of the most serious complications during operative hysteroscopy. The risk is related to the volume of irrigation fluid adsorbed by miometrial veins. The aim of our study is to assess the predictivity of a mathematical model built for extracellular volume monitoring.

Methods: We recruited 32 patients who underwent operative hysteroscopy with resectoscope at the university centers of Salerno. We measured plasmatic glucose levels, using fingerstick, blood pressure, serum sodium and potassium. These measurements were performed both before and after hysteroscopy. We also measured procedure duration and liquid balance. Data were processed by t-test and glycemia proved to be the most rapidly changing marker, in association with the irrigation fluid used. So, a mathematical model, based on forward stepwise regression analysis was built to predict glycaemia variations from the measurements of blood pressure, irrigation fluid balance and hysteroscopy duration.

Results: The proposed (∆Glicaemia=8.6539+0.2629*∆Diastolic pressure-0.3014*∆Sistolic pressure+0.5764*Duration+5.3195*∆Volume) model proved to be effective because predicted glycaemia was in line with the measured plasma glucose levels with a means difference below of the 10% of the differential value.

Conclusions: Since glycaemia is the most sensitive index of extracellular fluid composition and of the risk of intravasation, its prediction based on routinely monitored parameters during hysteroscopy (such as blood pressure, fluid balance and duration of the procedure) may be of help in intravasation risk assessment.

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Yield Potential and Adaptability of Selected Food-grade Sorghum Hybrids to Mediterranean Conditions

Sorghum has been grown as a food crop for many centuries in Africa and Asia and it is a cereal option for celiac patients. Due to its properties as a wheat-free food, interest is increasing in cultivating sorghum in Mediterranean countries. To evaluate the agronomic characteristics of growth in the Mediterranean area, six hybrids of sorghum (Sorghum bicolor L. Moench), representing different maturity classes, were selected and tested in experimental trials for 2 years in San Bartolomeo in Galdo (BN) hill, South Italy. The results showed moderate variation in adaptation of these hybrids as measured by differences in grain yields (4.60 to 7.30 t ha-1) and other pheno-morphological traits. In particular, the hybrid SW6129W from Bolivia and the hybrid SASG05W from Argentina had grain yields higher than others tested in these trials. Our results demonstrated that selected food-grade hybrids from South America, have agronomic characteristics well-suited for cultivation in Mediterranean countries.

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Gut microbiota composition and products contribute to gut–liver axis dysfunction in pediatric obesity related NAFLD, with distinct metabolomic signature

Gut microbiota (GM) and intestinal permeability (IP) are reported to interplay with wrong lifestyle in determining pediatric obesity (PO)/obesity-related complications. We concurrently investigated GM and gut-liver axis (GLA) components in 26 pre-pubertal obese children [OB(+)] with/without hepatic steatosis (13[ST(+)]/13[ST(−)]), vs 14 normal weight (NW) controls.

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The sexuological impact of hormonal contraceptives based on their route of administration

Evidence on the effects of hormonal contraceptives on female sexuality is conflicting. We enrolled 556 women, divided into six groups: two composed of subjects using a combined hormonal contraceptive (COC) containing 0.020 (“COC20”) and 0.030 (“COC30”) mg of ethynyl estradiol (EE), “natural”, using COC containing 1.5 mg of estradiol (E2), “ring”, using a vaginal ring releasing each day 0.015 mg of EE + 0.120 of etonogestrel, “subcutaneous”, using a progestin only subcutaneous contraceptive implant releasing etonogestrel and “controls”, using no hormonal contraceptive methods. The subjects were required to answer to the McCoy female sexuality questionnaire and were subjected to a blood test for hormonal evaluation. An ultrasound evaluation of the dorsal clitoral artery was also performed. The higher McCoy sexological value were recorded in the subdermal group; significant differences were recorded among the groups in terms of hormone distribution, with the higher levels of androstenedione in subdermal and control groups. The ultrasound evaluation of dorsal clitoral artery shows a significative correlation between pulsatility and resistance indices and orgasm parameters of McCoy questionnaire. The recorded difference in the sexual and hormonal parameters among the studied hormonal contraceptives may guide toward the personalization of contraceptive choice.

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