1,721,010 research outputs found
IRS-1 as a key point in the regulation of insulin-sensitivity in granulosa cells from PCOS women
No full textBackground: Polycystic Ovary Syndrome (PCOS) is a common endocrine disorder among women and is characterised by chronic low-grade inflammation, ovulatory dysfunction, hyperandrogenism, and often by insulin-resistance. However, little is really known on insulin/IGF signalling in PCOS ovaries. This study aim ed to investigate the amount of Insulin (IR) and IGF-I receptor type I (IGF-IR1) and their intracellular mediators in granulosa cells from PCOS women with respect to controls.
Methods: 28 women with PCOS [age: 34.8±4.2 yr; BMI: 25.4±5.5 kg/m2] diagnosed following the Rotterdam criteria, and 44 healthy controls [age: 37.7±4.3 yr; BMI: 23.4±4.3 kg/m2] were enrolled. Granulosa cells were isolated from follicular fluids and were lysed. Total protein content was quantified using a spectrophotometer. Total and phosphorylated insulin receptor (INSR) [pY1162/pY1163], total IGF1-receptor (IGF1R), total and phosphorylated AKT [pS473], total and phosphorylated ERK1 [pT202/pY204]- ERK2 [pT185/pY187], and phosphorylated IRS-1 [pS312] were measured by using specific ELISA kits. Moreover, due to the low amount of protein content obtained from each sample, a multiplex bead-based assay was performed in other 14 PCOS women and 14 controls in order to evaluate total and phosphorylated GSK3β [pS9], IRS1 [pS636], AKT [pS473], mTOR [pS2448], P70S6K [pT412], IR [pY1162/pT1163], PTEN [pS380], GSK3α [pS21], TSC2 [pS939], RPS6 [pS235/pS236], IGF1R [pY1135/pY1136], belonging to the Akt/mTOR pathway which is pivotal for insulin and IGF1R signaling. Student’s T-test and Fisher’s exact test were used to analyze the data.
Results: Phosphorylated IRS-1 was measurable in 9/21 PCOS patients and was undetectable in controls (P = 0.0002). Total and phosphorylated INSR, total IGF1R, total and phosphorylated AKT and total and phosphorylated ERK1-2 were similar in PCOS and controls. The multiplex analysis showed, however, that phosphorylated IGF1R (P = 0.02), IRS1 (P = 0.04), mTOR (P = 0.05), p70S6K (P <0.0001), INSR (P = 0.01), GSK3α (P = 0.02), and TSC2 (P = 0.01) were higher in PCOS with respect to controls. No differences were found in phosphorylated GSK3β, AKT, PTEN, RPS6 between PCOS and controls.
Conclusion: The phosphorylated fraction of IRS-1 observed in PCOS with both methodologies, methods suggests that IRS1 has a key role in the regulation of insulin-sensitivity in PCOS. Indeed, the increased phosphorylation at the Ser312 and Ser636 sites of IRS-1 is compatible with reduced insulin sensitivity.
Acknowledgments: This work was funded by the Italian Ministry of Health (grant number GR-2018-12367635 “Bando di Ricerca Finalizzata-Giovani Ricercatori 2018”)
Growth hormone (GH) deficiency and subsequent replacement therapy trigger differential expression of specific miRNAs in males and females: not just a matter of height
No full textIntroduction: Growth hormone (GH) is essential for stimulating growth and cell proliferation through its effects on metabolism, cartilage and bone growth. We have conducted initial studies to find new biomarkers for GH deficiency and early treatment response, focusing on miRNAs expressed in both sexes. We aimed at investigating sex-specific differences in circulating miRNAs at baseline and after 3 months on GH treatment in a cohort of prepubertal children with isolated idiopathic GH deficiency (IIGHD).
Methods: We re-analyzed our previously published miRNA global profiling (GSE193450) dataset to identify differences between females (F) (N:5, CA:7.44±2.46) and males (M) (N:5; CA:10.16 ± 2.14) before and after 3 months of GH therapy. Significant miRNAs were selected based on p-value (P < 0.05) or fold change (log22−DDCt) > +1.5 or FC (log22−DDCt) < −1.5. KEGG enrichment was performed with deregulated miRNAs to establish biological pathways impacted.
Results: At baseline, we identified 13 differentially expressed miRNAs between F and M. The 4 miRNAs upregulated in females (hsa-miR-380-3p, hsa-miR-486-5p, hsa-miR-325, hsa-miR-185-5p) and the 9 upregulated miRNA in males (hsa-miR-30b-5p, hsa-miR-30c-5p, hsa-miR-132-3p, hsa-miR-139-5p, hsa-miR-142-5p, hsa-miR-195-5p, hsa-miR-197-3p, hsa-miR-200c-3p, hsa-miR-340-5p) had a similar effect on the cell cycle pathway and Wnt signaling, involved in osteoblast differentiation and bone metabolism. However, miRNAs in F had a greater impact on p53 signaling, crucial for osteogenic differentiation, whereas miRNAs in M affected mostly TGF-β signaling, involved in bone formation. After 3 months on GH therapy, two of the miRNA above (hsa-miR-325 and hsa-miR-30c-5p) were still differentially expressed in M and F, suggesting that hormone replacement didn't restore sex-specific differences. Other 8 miRNAs were differentially expressed in F and M after 3 months of therapy and were involved in cell cycle, p53, mTOR (mediator of bone anabolism) and insulin signaling. Nevertheless, the level of significance of these pathways was different between the two groups. With an analysis comparing baseline and 3 months of therapy in M and F separately, we identified that cell cycle was similarly affected by deregulated miRNAs in both sexes. However, p53 and insulin signaling, and endocytosis were more impacted in F whereas pathways in M were enriched for Wnt signaling and RNA transport.
Conclusion: This study evidenced differences in miRNAs expression profiles in GH deficient males and females, both at baseline and after 3 months on GH therapy. Our aim is to emphasize the importance of considering gender-specific factors in therapeutic strategies, which currently has not been done for GH deficiency
COVID-19 stressful conditions in the increase of central precocious puberty: long-term follow-up and timelines of this phenomenon demonstrate the declines to pre-pandemic levels after the end of online classes period in Italy
No full textIntroduction: COVID-19 pandemic resulted in serious social challenges and physical and mental repercussions. In many countries, central precocious Puberty (CPP) increased dramatically but long-term data are really reduced.
Aim: A retrospective evaluation of incidence and characteristics of CPP before and during COVID-19 pandemic in four Italian Paediatric Hospitals (Florence, Modena, Reggio-Emilia and Parma).
Patients and methods: 449 patients diagnosed by 01 January 2018 to 31 December 2022 were collected. We divided this period in pre-COVID-19 (pre-Cov19: from 01/01/2018 to 08/03/2020) and COVID-19 period (Cov19: from 08/03/2020 to 31/12/ 2022); Cov19 period was further divided into lockdown (lock-Cov19), restrictions (restr-Cov19), post-restriction (postrestr-Cov19) periods.
Results: During pre-Cov19, 111 CPP (7.8 ± 0.6 years) were diagnosed (4.3 cases for month); of these, 7.2% showed a rapidly progressive course (RPPP) and 14.4% were untreated CPP disclosing a RPPP during the lockdown. During Cov19, 338 CPP (7.8 ± 0.9 years) were diagnosed (10.0 cases for month; P < 0.05); among Cov19, we diagnosed 9.8 cases for month (P < 0.05) during lock-Cov19 (7.4 ± 1.2; P < 0.005), 14.4 cases for month (P < 0.0001) during restr-Cov19 (7.7 ± 1.0; P = NS), and 5.4 cases for month (P = NS) during postrestr-Cov19 (7.8 ± 1.0: P = NS). In the Cov19 period 36.7% of CPP were RPPP: 55.5% in lock-Cov19 (P < 0.0001), 36.7% in restr-Cov19 (P < 0.0001) and 8.8% in postrestr-Cov19 (P = NS). Basal LH (BLH), peak LH (PLH), DPLH/BLH, ovary volume and uterine length were significantly different in lock-Cov19 and restr-Cov19, but not postrestr-Cov19, in respect to pre-Cov19. BMI, bone age and height SDS were not different among these groups.
Conclusion: CPP diagnosis significantly increase during the first year of the Covid-19 pandemic in respect to pre-pandemic years and post-restriction year; CPP incidence decrease particularly after the end of online classes at home, stressing the use of electronic devices as one of the main causes of CPP during the COVID-19 pandemic
Docosahexaenoic acid (DHA) is reduced and could be protective against Hashimoto’s thyroiditis in children with Down syndrome: a cross-sectional study
No full textInflammation is a known feature of Down syndrome (DS) and is caused by a dysregulation between pro and anti-inflammatory cytokines. Hashimoto's thyroiditis (HT) is characterised by a slowly developing persistent inflammation of the thyroid gland which frequently leads to hypothyroidism. In DS children, HT is the most common autoimmune disease ad its prevalence has been reported to be more elevated than that generally seen in age-matched patients without DS: 34% vs 1.3%, respectively. The diagnosis of HT is based on dysthyroidism and/or presence of antibodies against thyroid peroxidase and thyroglobulin, although seronegative forms can be seen in 5%–10% of cases. The ultrasound features of HT include decreased echogenicity, heterogeneity, hypervascularity, and presence of small cysts. Adequate blood levels of omega-6 (linoleic and arachidonic acid) and omega-3 (EPA and DHA) polyunsaturated fatty acids (PUFAs) may play a protective role against inflammation as they inhibit leukocyte chemotaxis and pro-inflammatory cytokine production. In this cross-sectional study we investigated DHA levels in a case series of pediatric DS patients and explored any correlations with HT. Twenty-six patients with chromosome 21 free trisomy karyotype aged 0-18 years were enrolled. Patients with congenital and isolated subclinical hypothyroidism were excluded. For each patient, a capillary blood sample was analyzed using the "Nexis GC-2030 gas chromatography" method to verify the PUFAs profile. Thyroid function (TSH, FT4, and anti-thyroid antibodies) and the presence of ultrasound signs of HT were assessed in all patients at the time of sampling. Clinical symptoms and/or signs of HT on ultrasound were present in 10/26 subjects (38%; 60% F) who had positive antibodies against thyroid peroxidase and thyroglobulin also, and required Levo-thyroxine replacement therapy (mean age 9 yr ± 68 mo). One subject had ultrasound signs of thyroiditis without circulating antibodies and was euthyroid, and further 2 subjects (8%) had hyperthyroidism at onset of HT requiring treatment with Methimazole. Mean DHA percentage was 1.5% (0.34-2.86), in females (N: 14) 1,37% (0.34-2.58) and in boys (N: 12) 1.6% (0.61-2.86). No significant differences related to age and BMI. DHA levels were negatively correlated with the presence of HT (95% confidence interval 0.67-1.56; t-test P <0.001) in the entire population. HT was confirmed to be a frequent endocrinopathy in DS and was associated with lower DHA levels compared with euthyroidism and absence of autoimmunity. We hypothesize that DHA intake could play a protective anti-inflammatory role by preventing or delaying the onset of HT in DS
The LIFE-MILCH study: first data on the exposure to Endocrine Disrupting Chemicals (EDCs) in urine and breast milk (BM) from end of pregnancy to 12 months of life
No full textIntroduction: The ongoing LIFE-MILCH project (www.lifemilch.eu) focuses on detecting EDCs in mothers, in BM and in urine, and in infants from birth up to 12 months of age studying relationships with neurodevelopment, growth, distribution of adiposity, pubertal stages, ano-genital distances, life-style and professional sources of exposure (questionnaires) to establish a risk assessment model to prepare safety guidelines to optimize all benefits related with breastfeeding and preserve future health.
Objective: To evaluate exposure to EDCs in mother’s and infant’s urine and in BM.
Methods: These preliminary data are relative to 200/654 mother-infant dyads enrolled in Parma, Reggio Emilia and Cagliari, in Italy. Urine samples were collected and analyzed at recruitment/birth (T0), 1 (T1), 3 (T2) and 6 months (T3) in both the mothers and infants. BM was collected at T1, T2 and T3. In all biological samples bisphenol (BP) A, BPS, BPF, phthalates (PHTs) and their metabolites (dibutyl phthalate (DBP), benzyl butyl phthtalate (BBP), di-(2-ethylhexyl) phthalate (DEHP), monobutyl phthtalate (MBP), monobenzyl phthalate (MBzP), mono(2-ethylhexyl) phthalate (MEHP), mono(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP), mono(2-ethyl-5-oxyhexyl) phthalate (MEHOP)), parabens (PBs) (butylPB, isobutylPB), pesticides (glyphosate, aminomethylphosphonic acid, glufosinate) polycyclic aromatic hydrocarbons(PAHs), pyrethroids insecticides, and heavy metals were measured by LC-MS.
Results: BPs were detected in 85,7% of the mother’s samples at T0, in 96,7% at T1, in 93,4% at T2. BPA was ubiquitous in mother’s urine and milk, at all times, and was detected in 32,1% of newborn’s urine, in 55,4% at T1, and increased at T2 and T3. BPS was detected in mother’s and infant’s urine and in BM at all times; BPS was detected in the newborns. Among PHTs, DBP was detected in 80% of BM samples at all times. MBP was detectable in 80% of mother’s urine samples at T0, and above 90% at all other times. BBP was detectable in BM only in 24,2% of samples at T1 and in 53,3% at T3. DEHP metabolites were found in urine and BM at all times. MBP was detectable in 85,4% of infant’s urine samples at birth and findings were similar at the following time points. PBs were almost absent in mother’s samples, but were detectable in infant’s urine samples and iBuPB reached a detection rate of 53,3% at T2. Heavy metals were not found.
Conclusion: EDCs are present in BM, maternal and infant’s urine samples with some differences in exposure. Increasing awareness and prevention campaigns are of utmost importance
Going Beyond Counting First Authors in Author Co-citation Analysis
Get PDFThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
MRI-based radiomics of the pituitary gland is highly predictive of CentralPrecocious Puberty in girls: pilot study
No full textIntroduction: The diagnostic gold standard for Central Precocious puberty (CPP) is the gonadotropin-releasing hormone (GnRH) stimulation test. MR imaging of the brain (MRI) and the hypothalamus-pituitary region is required to exclude organic causes.
Objective: The aim of the study was to explore a radiomic model that could assist physicians in the diagnostic workup of CPP.
Methods: 45 girls with a confirmed diagnosis of CPP and 47 age-matched pre-pubertal female subjects (control group) were retrospectively enrolled. Two readers (R1, R2) with different levels of expertise on pediatric neuroradiology blindly segmented the pituitary gland on MRI studies for radiomic features (RFs) calculation and performed a manual estimation of pituitary volume (ellipsoid approximation - EA). Cross-validated linear discriminant analysis was used to develop for each reader both a radiomic model and a clinical reference model based on the manually-measured pituitary volume. Radiomics was compared against the EA in terms of: 1) predictive performances (metrics: ROC-AUC, accuracy, sensitivity and specificity); 2) reliability of predictors between readers (metric: intraclass correlation coefficient, ICC); 3) consistency of performance metrics between readers (absolute difference between R1 and R2 mean performances).
Results: The radiomic model significantly improved the diagnostic sensitivity with respect to the EA of the pituitary gland (0.78 versus 0.69 in validation set, P <0.001) and achieved a well-balanced trade-off between sensitivity and specificity. Radiomic predictors demonstrated higher inter-reader reliability (ICC>0.57) with respect to the EA predictor (ICC=0.46). Moreover, the radiomic model showed a greater consistency between R1 and R2 findings, with a mean difference among all performance metrics of 0.06±0.04, which was lower than the difference observed for EA (0.10±0.02).
Conclusion: Radiomics of the pituitary gland alone demonstrated a greater potential in diagnosing CPP and an increased reliability with respect to EA. This opens a way to diagnosing CPP based on MRI of the pituitary gland in addition to clinical and hormonal data. However, further studies are warranted to validate these preliminary data
A comprehensive overview of the changes in IGF system peptides in the follicular fluid (FF) of women with Polycystic Ovarian Syndrome (PCOS) and their relationships with BMI and HMGB1
No full textBackground: PCOS is characterised by chronic low-grade inflammation, ovulatory dysfunction, hyperandrogenism, and insulin-resistance. The IGFsystem includes IGF-I, -II and seven IGFBPs, which regulate IGFbioavailability. Chronic inflammation modifies the IGF system that regulates ovarian function and glucose metabolism. HMGB1 is related with both inflammation and insulin sensitivity;we previously described increased HMGB1 FF in PCOS. This study aim ed to investigate the IGFsystem in PCOS.
Methods: 70 women with PCOS [age: 34.1±4.7yr; BMI: 25.6±5.6 kg/m2] diagnosed following the Rotterdam criteria, and 70 healthy controls [age: 36.8±3.8yr; BMI: 24±5 kg/m2] were enrolled. Subjects were stratified based on BMI <25 (underweight/normal weight) or ≥25 (overweight/obese). Induction of follicular development for IVFwas conducted according to a long luteal GnRH agonist protocol. FF were collected during oocyte retrieval and centrifuged to remove red blood cells and debris. IGF-I, IGF-II, IGFBP-1-7, and HMGB1 were measured in FF using specific ELISAkits. The concentrations were converted to nM and bioactivity was calculated as ratios between IGFs and IGFBPs. Student’sT-test, ANOVA and Pearson’s correlation were used to analyze the data.
Results: IGF-II (387.2±210.5 vs 495.7±157.9 ng/ml, P = 0.0007) and IGFBP-4 (16.5±7.7 vs 20.7±9.5 ng/ml; P = 0.005) were lower whereas IGFBP-6 (55570±27793 vs 46417±22110ng/ml; P = 0.03) and IGFBP-7 (405.9±211.6 vs 324.1 ± 145.8ng/ml; P = 0.009) were increased in PCOS compared with controls. IGF-I, IGFBP-1, -2, and -3 were similar in both groups. IGFBP-5 was undetectable. HMGB1 was increased in PCOS (41.5±22.1 vs 29.5±20.4ng/ml; P = 0.002). Interestingly, stratification based on BMI showed that IGF-II (380.7±201.8 vs 514.1±165.3ng/ml P = 0.005) and IGFBP-4 (15.2±6.9 vs 20.0 ± 9.7ng/ml, P = 0.04) were lower whereas IGFBP-7 (444.7±251.5 vs 300.6±103.6ng/ml P = 0.001) was increased in underweight/normalweight PCOS with respect to underweight/normal weight controls. IGFBP-2 was reduced in the overweight/obese subjects in both controls and PCOS, and was negatively correlated with BMI in both groups. IGFBP-3 was correlated with IGFBP-4 (r =+0.45, P <0.0001) in controls, and HMGB1 was correlated with IGFBP-2 (r =+0.34, P = 0.007) in PCOS. When considering the molar ratios, IGF-II bioavailability was confirmed to be significantly lower in PCOS whereas IGF-I bioavailability was unchanged.
Conclusion: The reduction of IGF-II and IGFBP-4 in PCOS is compatible with reduced follicular development. The changes in IGF-II, IGFBP-4, and IGFBP-7in the underweight/normalweight PCOS suggests that these are specific to PCOS and unrelated to weight changes, although obesity determines changes in FF(IGFBP-2). The correlation between HMGB1 and IGFBP2 is compatible with the increased inflammatory status in PCOS. The increase of IGFBP-6 and -7opens the way for further understanding of the pathogenesis of PCOS. This work was funded by the Italian Ministry of Health (grant number GR-2018-12367635”)
The LIFE-MILCH project: preliminary data from the risk assessment model of exposure to Endocrine Disrupting Chemicals (EDCs) in mother-infant dyads during the first 3 months of life
No full textIntroduction: The ongoing LIFE-MILCH project (www.lifemilch.eu), focuses on detecting EDCs in mothers, in breast milk (BM) and in urine, and in infants from birth up to 12 months of age studying relationships with neurodevelopment, growth, distribution of adiposity, pubertal stages, and ano-genital distances, life-style sources (questionnaires) of exposure to establish a risk assessment model to prepare safety guidelines to optimize all benefits related with breastfeeding and preserve future health.
Objective: To estimate the risk of exposure to EDCs for infants especially through BM.
Methods: The current preliminary risk assessment model regards the data of approximately 200/654 mother-infant dyads, enrolled at 3 sites in Italy. Healthy mothers were enrolled at 36-41 weeks of gestational age, pregnancies were uncomplicated. Urine samples were collected in mothers and infants at enrolment(T0) and at 1(T1), 3(T2) and 6 months(T3) after delivery. BM samples were collected at T1, T2, T3. In all biological samples bisphenol (BP)A, BPS, BPF, phthalates (PHTs) and their metabolites (DBP, BBP, DEHP, MBP, MBzP, MEHP, MEHHP, MEHOP), parabens (PBs) (BuPB, iBuPB), pesticides (glyphosate, aminomethylphosphonic acid, glufosinate) polycyclic aromatic hydrocarbons (PAHs), pyrethroids insecticides, and heavy metals were measured by LC-MS.
Results: When BPs were detected in mothers’urine, it was more likely to detect these also in the infants’ urine samples, especially at T0(OR=7.06). Exclusive breastfed infants at T1 and at T2 were 3 and 5 times more likely to have at least one BPs in urine at T1 and T2, respectively. At T0, mothers with detectable PHTs in urine had a higher probability to have children with these molecules in their urines(OR=4.02 for PHTs and 11.5 for PTHs metabolites). At T1, detectable PTH metabolites in BM increased the probability of finding these molecules in the children’s urines (OR=8.95). Overall, if PBs were found in BM there was a 32-fold higher probability of detecting these in the childrens’urine samples at T2. When pesticides were found both in BM and in the urine of mothers, the probability of finding these compounds in the urine of their infants was increased (OR=3.27 for BM at T2). The presence of PAHs in mothers’urine and BM doubled the probability of finding PAHs in the infants’urine at T2.
Conclusion: These data confirm exposure related with a shared environment, and the passage of EDCs from mothers to infants besides children exposure. The exposure to these EDCs has to be reduced for the health of our children
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