Urinary (1)H-NMR-based metabolic profiling of children with NAFLD undergoing VSL#3 treatment.

BACKGROUND: Nowadays, non-alcoholic fatty liver disease (NAFLD) is one of the most common chronic liver diseases in children. Our recent clinical trial demonstrated that dietary and VSL#3-based interventions may improve fatty liver by ultrasound and body mass index (BMI) after 4 months. OBJECTIVES: As in this short-term trial, as in others, it is impracticable to monitor response to therapy or treatment by liver biopsy, we aimed to identify a panel of potential non-invasive metabolic biomarkers by a urinary metabolic profiling. METHODS: Urine samples from a group of 31 pediatric NAFLD patients, enrolled in a VSL#3 clinical trial, were analyzed by high-resolution proton nuclear magnetic resonance spectroscopy in combination with analysis of variance-Simultaneous Component Analysis model and multivariate data analyses. Urinary metabolic profiles were interpreted in terms of clinical patient feature, treatment and chronology pattern correlations. RESULTS: VSL#3 treatment induced changes in NAFLD urinary metabolic phenotype mainly at level of host amino-acid metabolism (that is, valine, tyrosine, 3-amino-isobutyrate or ß-aminoisobutyric acid (BAIBA)), nucleic acid degradation (pseudouridine), creatinine metabolism (methylguanidine) and secondarily at the level of gut microbial amino-acid metabolism (that is, 2-hydroxyisobutyrate from valine degradation). Furthermore, some of these metabolites correlated with clinical primary and secondary trial end points after VSL#3 treatment: tyrosine and the organic acid U4 positively with alanine aminotransferase (R=0.399, P=0.026) and BMI (R=0.36, P=0.045); BAIBA and tyrosine negatively with active glucagon-like-peptide 1 (R=-0.51, P=0.003; R=-0.41, P=0.021, respectively). CONCLUSIONS: VSL#3 treatment-dependent urinary metabotypes of NAFLD children may be considered as non-invasive effective biomarkers to evaluate the response to treatment.

First trimester serum neutrophil gelatinase-associated lipocalin in gestational diabetes.

AIMS: Neutrophil gelatinase-associated lipocalin (NGAL) is highly expressed in damaged epithelia, during inflammation and cardiovascular disease. Soluble NGAL was measured in women who subsequently developed gestational diabetes. METHODS: From a cohort of 908 pregnant outpatients who participated in a screening programme for Down's syndrome at 9-12 weeks of gestation, we considered all 41 women with a singleton pregnancy, who developed gestational diabetes in the previous 12 months, and a control group of 82 normal pregnancies. Circulating NGAL and insulin resistance by homeostasis model assessment ratio (HOMA-IR) were determined in the first trimester. RESULTS: Median serum NGAL concentrations were higher in the gestational diabetes group [51.3 ng/ml (39.8-66.1 ng/ml)] compared with the control group [17.8 ng/ml (15.5-20.9 ng/ml)] (P < 0.001) and positively correlated with HOMA-IR (P < 0.001). CONCLUSIONS: In the first trimester, circulating NGAL was significantly increased in women who subsequently developed gestational diabetes compared with the control group.

Endocrine changes in postmenopausal women after high-dose danazol therapy.

BACKGROUND: Purpose of the study was to investigate changes in the hormonal pattern in postmenopausal women after high dose danazol therapy. In 16 postmenopausal women (mean age 66.4 +/- 8.6) treated with danazol 1.2 g/die x21 we studied serum levels, before and after therapy, of FSH, LH, E2, fE2, T, fT, D4-A, DHEAS, SHBG. METHODS: FSH, LH, SHBG by IRMA kits (DPC); E2, T, DHEAS, D4-A by RIA kits (Sorin Biomedica) after celite microcolumn chromatography; fT by coat-A-count kit (DPC); fE2 by equilibrium dialysis; serum albumin by bromocresol-green colorimetric methods. RESULTS: After therapy there was a statistically significant decrease of LH by 13%, of FSH by 19%, SHBG by 84% and increase of fE2 by 24.5%, fT by 77%. Other hormones show changes that are not statistically significant. CONCLUSIONS: In postmenopausal women danozol therapy, even if at a higher dose, has about same effect on hormonal pattern as in normally cycling women. Danazol showed a clear antigonadotropic effect and can play a role in therapy when reduction of gonadotrophins and E2 secretion is desirable.

The clinical utility of serum uric acid measurements in pre-eclampsia and transient hypertension in pregnancy.

BACKGROUND: Our purpose was to evaluate the clinical utility of serum uric acid measurements in the hypertension diseases of pregnancy. METHODS: We identified 286 women and categorized them into three diagnostic groups according to definitions of hypertensive diseases in pregnancy published by the National Working Group on Hypertension in Pregnancy: pre-eclampsia (94), transient hypertension (102) and normal (90). We compared the median uric acid concentration for each group and calculated the sensitivities and the specificities in diagnosing pre-eclampsia. The results were analyzed by the Mann-Whitney test. RESULTS: Median serum uric acid values in the pre-eclamptic group, in the transient hypertension group and in the control group were 375 (262-536) mumol/L, 309 (214-387) mumol/L, 259 (143-339) mumol/L, respectively. Compared with normal, the median serum uric acid levels in women with pre-eclampsia or transient hypertension were significantly elevated. Differences in median serum uric acid concentrations between women with preeclampsia and with transient hypertension were statistically significant too. The prevalence of IUGR in the pre-eclamptic group and transient hypertension group was 65.9% and 29.4%, respectively. Sensitivity for serum uric acid levels of 339 mumol/L was 77.3% in the pre-eclamptic group and 32.3% in the transient hypertension group; the difference was statistically significant. Specificity was exactly the same in both groups (92%). CONCLUSIONS: Our data, in accordance with international literature, confirm the clinical utility of serum uric acid as a marker of pre-eclampsia, but not of transient hypertension. Furthermore its high predictive value makes it possible to select a group of pre-eclamptic women with high risk for intrauterine growth retardation.

Placental growth hormone in Down's syndrome screening.

A number of serum markers have been proposed to improve the sensitivity (and specificity) of the triple test, which, until now, has been the gold standard in second-trimester serum screening for Down's syndrome. Among them, human placental growth hormone (hPGH) has been proposed because of its significantly elevated serum levels in pregnancies affected by chromosomal aneuploidies. Our experience, on maternal serum stored from 32 Down's syndrome-affected pregnancies, confirms a slight but significant increase in hPGH levels compared with controls. These data summarized to that of the previous screening could give a calculated detection rate of 71.9%, better than that of the standard triple test alone (65.6%).

Hormonal disregulation and catamenial epilepsy.

Six epileptic women, showing a clear increase in seizure frequency in the perimenstrual period, were studied during three consecutive menstrual cycles. At each control seizure occurrence, EEG recordings, endocrine profile and antiepileptic drug plasma levels were evaluated. The only pathological finding was the progesterone deficiency with a relative hyperestrogenism during the late luteal phase. Our data agree with other reports suggesting that catamenial epilepsy may result from a sex hormone disregulation.

[Serum Ca-125 levels in aneuploidy screening in first pregnancy trimester. Authors' experience]. / Valore del Ca-125 nello screening delle aneuploidie nel primo trimestre di gestazione. Nostra esperienza.

BACKGROUND: The aims of the current study were to investigate the value of Ca-125 maternal serum levels at the first trimester of pregnancy in order to test heir usefulness for the screening of foetal aneuploidies. MATERIALS AND METHODS: The study was performed retrospectively on frozen maternal serum samples obtained in the antenatal clinics of the department of obstetrics and gynaecology: 85 control normally cycling women, 479 pregnant over 36 years from 8th to 12th week of pregnancy underwent the villocentesis, 383 pregnant women aged from 19 to 45 years underwent the "tritest" (AFP = alpha-fetoprotein, hCG = human chorionic gonadotropin, fE3 = free estriol, age) for the screening of the Down syndrome in the second trimester of pregnancy. All newborns have been tested by Apgar score. The immunoradiometric (IRMA) Ca-125 assay was performed without knowing the result of the cytogenetic test. Over all we have had 11 patients with aneuploidy, 7 Down, 3 Edwards and 1 Patau. The levels of Ca-125 are expressed as MoM multiple of median) in order to compare the levels at different gestational age. RESULTS: We had no significant differences between normal and pathological pregnancies, therefore we don't reckon the Ca-125 assay useful in the screening of aneuploidies in the first trimester of pregnancy.

Gonadotropinic reserve in women affected by hyperprolactinemic amenorrhea.

The Authors have evaluated the hypophysis gonadotropic reserve after stimulation by synthetic GnRH (100 ng) in women affected by functional hyperprolactinemia and by prolactin-secreting adenoma. The LH response was significatively higher in women affected by functional hyperprolactinemia. The PRL values seem to have no influence on the gonadotropic reserve except for the cases in which The PRL levels were higher than 200 ng/ml. In all these cases a prolactinoma was present. The FSH response was similar in the two groups considered.

Randomised clinical trial: The beneficial effects of VSL#3 in obese children with non-alcoholic steatohepatitis.

BACKGROUND: Gut microbiota modifiers may have beneficial effects of non-alcoholic fatty liver disease (NAFLD) but randomised controlled trials (RCT) are lacking in children. AIM: To perform a double-blind RCT of VSL#3 vs. placebo in obese children with biopsy-proven NAFLD. METHODS: Of 48 randomised children, 44 (22 VSL#3 and 22 placebo) completed the study. The main outcome was the change in fatty liver severity at 4 months as detected by ultrasonography. Secondary outcomes were the changes in triglycerides, insulin resistance as detected by the homoeostasis model assessment (HOMA), alanine transaminase (ALT), body mass index (BMI), glucagon-like peptide 1 (GLP-1) and activated GLP-1 (aGLP-1). Ordinal and linear models with cluster confidence intervals were used to evaluate the efficacy of VSL#3 vs. placebo at 4 months. RESULTS: At baseline, moderate and severe NAFLD were present in 64% and 36% of PLA children and in 55% and 45% of VSL#3 children. The probability that children supplemented with VSL#3 had none, light, moderate or severe FL at the end of the study was 21%, 70%, 9% and 0% respectively with corresponding values of 0%, 7%, 76% and 17% for the placebo group (P < 0.001). No between-group differences were detected in triglycerides, HOMA and ALT while BMI decreased and GLP-1 and aGLP1 increased in the VSL#3 group (P < 0.001 for all comparisons). CONCLUSIONS: A 4-month supplement of VSL#3 significantly improves NAFLD in children. The VSL#3-dependent GLP-1 increase could be responsible for these beneficial effects. Trial identifier: NCT01650025 (www.clinicaltrial.gov).

Adiponectin and insulin resistance in early- and late-onset pre-eclampsia.

OBJECTIVE: To evaluate the importance of adiponectin and insulin resistance in early- and late-onset pre-eclampsia. DESIGN: A nested case-control study in 72 pregnant women who participated in the first-trimester Down-syndrome-screening programme and who delivered at our hospital. SETTING: University Hospital, Department of Obstetrics and Gynecology. POPULATION: Pregnant women: 36 women with pre-eclampsia of which 20 late onset and 16 early onset were compared with 36 uncomplicated pregnancies who delivered at term. METHODS: In all the women, insulin resistance was calculated by the homeostasis model assessment ratio (HOMA-IR) and plasma adiponectin was determined using an enzyme-linked immunosorbent assay. MAIN OUTCOME MEASURES: Insulin resistance and adiponectin concentration. RESULTS: First-trimester plasma adiponectin mean levels in the whole pre-eclampsia group were significantly lower than that in the control group (8.4 +/- 3.3 versus 14.8 +/- 4.6 microgram/ml; P < 0.001), whereas first-trimester mean HOMA-IR values were significantly higher in the pre-eclampsia group than that in the control group (2.0 +/- 1.1 versus 1.0 +/- 0.4; P= 0.01). Plasma adiponectin concentrations at delivery in the pre-eclampsia group were significantly higher than that in the control group (9.2 +/- 3.7 versus 7.8 +/- 2.6 microgram/ml; P= 0.04). First-trimester plasma adiponectin mean concentrations in the late-onset subgroup were significantly lower compared with the concentrations in early-onset subgroup (6.2 +/- 1.4 microgram/ml versus 11.1 +/- 3.2 microgram/ml; P < 0.001), and there was a significant difference in adiponectin plasma values only between women in the late-onset pre-eclampsia group versus those in the control group (P < 0.001). First-trimester mean HOMA-IR values were significantly higher in the late-onset subgroup compared with that of the early-onset subgroup (2.5 +/- 1.3 versus 1.3 +/- 0.3; P= 0.02), and there was a significant difference only between the control group versus the late-onset subgroup (P= 0.001). CONCLUSIONS: First-trimester adiponectin and HOMA-IR values seem to select two completely different populations: early- and late-onset pre-eclampsia, which might suggest a different pathogenesis.

4-hour urinary and serum immunofluorimetric assay for luteinizing hormone to detect onset of preovulatory LH surge.

The initial surge of LH is an important preovulatory event. The validity of a time-resolved immunofluorimetric assay method for determining LH both on the samples of serum and urine are reported in our study. A good correlation exists between the serum and urinary LH with the presence of a shorter interval in the length of the LH increase in the urinary dosage in respect to the serum dosage. There emerges when comparing the RIA and IFMA techniques that a good correlation exists both in the urinary and in the serum samples. However the values found with the IFMA method are on the average inferior (ca. 50%) in respect to those obtained with the RIA technique.

Determination of 17-hydroxyprogesterone in the normal and induced ovulatory cycles.

Concentrations of 17-hydroxyprogesterone (170HPg) were measured in serum samples obtained daily through cycles both spontaneous and induced by gonadotropins. These samples had previously been assayed for estradiol, progesterone, luteinizing hormone (LH). The concentration of 170HPg increases considerably the day before the LH peak in the ovulatory cycles. In the anovulatory cycles the 170HPg levels are shown as being low or the constant high levels in one case has not been followed by the LH peak. In our opinion the parameter taken into consideration does not lend itself completely to the prediction of the ovulatory event but in synchronism with the LH peak, may be a useful index of the follicular luteinization both in the spontaneous ovulatory cycles and in the stimulated cycles where the estrogenic increase is abnormal.