RAGE inhibition blunts insulin-induced oncogenic signals in breast cancer.

The receptor for advanced glycation end products (RAGE) is implicated in diabetes and obesity complications, as well as in breast cancer (BC). Herein, we evaluated whether RAGE contributes to the oncogenic actions of Insulin, which plays a key role in BC progression particularly in obese and diabetic patients. Analysis of the publicly available METABRIC study, which collects gene expression and clinical data from a large cohort (n = 1904) of BC patients, revealed that RAGE and the Insulin Receptor (IR) are co-expressed and associated with negative prognostic parameters. In MCF-7, ZR75 and 4T1 BC cells, as well as in patient-derived Cancer-Associated Fibroblasts, the pharmacological inhibition of RAGE as well as its genetic depletion interfered with Insulin-induced activation of the oncogenic pathway IR/IRS1/AKT/CD1. Mechanistically, IR and RAGE directly interacted upon Insulin stimulation, as shown by in situ proximity ligation assays and coimmunoprecipitation studies. Of note, RAGE inhibition halted the activation of both IR and insulin like growth factor 1 receptor (IGF-1R), as demonstrated in MCF-7 cells KO for the IR and the IGF-1R gene via CRISPR-cas9 technology. An unbiased label-free proteomic analysis uncovered proteins and predicted pathways affected by RAGE inhibition in Insulin-stimulated BC cells. Biologically, RAGE inhibition reduced cell proliferation, migration, and patient-derived mammosphere formation triggered by Insulin. In vivo, the pharmacological inhibition of RAGE halted Insulin-induced tumor growth, without affecting blood glucose homeostasis. Together, our findings suggest that targeting RAGE may represent an appealing opportunity to blunt Insulin-induced oncogenic signaling in BC.

What factors have impact on glucocorticoid replacement in adrenal insufficiency: a real-life study.

PURPOSE: The impact of patient's characteristics on glucocorticoid (GC) replacement therapy in adrenal insufficiency (AI) is poorly evaluated. Aims of this study were to assess the influence of sex and body weight on GC dosing and to describe the choice of GC in AI of different etiologies. METHODS: We retrospectively evaluated hydrocortisone (HC) equivalent total daily dose (HC-TDD) and per-kg-daily dose (HC-KDD) in 203 patients (104 primary AI [pAI], 99 secondary AI [sAI]) followed up for ≥ 12 months. They were treated with HC, modified-release HC (MRHC) or cortisone acetate (CA) and fludrocortisone acetate (FCA) in pAI. RESULTS: At baseline, CA was preferred both in pAI and sAI; at last visit, MRHC was most used in pAI (49%) and CA in sAI (73.7%). Comparing the last visit with baseline, in pAI, HC-TDD and HC-KDD were significantly lower (p = 0.04 and p = 0.006, respectively), while FCA doses increased during follow-up (p = 0.02). The reduction of HC-TDD and HC-KDD was particularly relevant for pAI women (p = 0.04 and p = 0.002, respectively). In sAI patients, no change of HC-KDD and HC-TDD was observed, and we found a correlation between weight and HC-TDD in males (r 0.35, p = 0.02). CONCLUSIONS: Our real-life study demonstrated the influence of etiology of AI on the type of GC used, a weight-based tailoring in sAI, a likely overdosage of GC treatment in pAI women at the start of treatment and the possibility to successfully increase FCA avoiding GC over-treatment. These observations could inform the usual clinical practice.

Pathogenesis and inflammatory response in experimental caprine mastitis due to Staphylococcus chromogenes.

Coagulase-negative staphylococci (CNS) are the most frequently isolated bacteria in cases of subclinical mastitis in dairy cows. CNS species may differ in their pathogenicity, but very little is known about their virulence factors or their immune response in intramammary infections. To our knowledge, no experimental studies into the mastitis pathogenesis caused by CNS have been described in lactating goats. The aim of this study was to induce an experimentally Staphylococcus chromogenes mastitis in lactating goats aimed at verifying if the model can be used to evaluate the inflammatory response, the dynamics of infection and the pathological findings within the first hours of intramammary inoculation. Six Saanen goats in mid-lactation were inoculated with 1 × 107 colony forming units of S. chromogenes. Bacterial growth peaked in milk from the challenged right halves of the mammary glands (RMG) at 4 h post inoculation (PI). Shedding of viable bacteria showed a marked decrease at 12 h PI. An increase in mean somatic cell counts was observed in the milk samples from 8 h PI onwards. Mild clinical signs were evoked by intramammary inoculation. Staphylococcus chromogenes could be isolated in tissue from all RMG. Histological examination of specimens of the RMG and lymph nodes of the goats showed an increased inflammatory response throughout the experiment with respect to control halves. In conclusion, the experimental inoculation of S. chromogenes in lactating goats is capable of eliciting an inflammatory response and capable of causing pathological changes. This research represents a preliminary study for a better knowledge of the mastitis pathogenesis caused by S. chromogenes.

Maternal serum PAPP-A and free ß-hCG at 12, 22 and 32 weeks' gestation in screening for pre-eclampsia.

OBJECTIVE: To examine the distribution of maternal serum pregnancy-associated plasma protein-A (PAPP-A) and free ß-human chorionic gonadotropin (ß-hCG) at 12, 22 and 32 weeks' gestation in singleton pregnancies which develop pre-eclampsia (PE) and examine the performance of these biomarkers in screening for PE. METHODS: Serum PAPP-A and free ß-hCG were measured in 94 989 cases at 11-13 weeks, 7597 at 19-24 weeks and 8088 at 30-34 weeks' gestation. Bayes' theorem was used to combine the a-priori risk from maternal characteristics and medical history with PAPP-A and free ß-hCG. The empirical and model-based performance of screening for preterm PE requiring delivery < 37 weeks' gestation and term PE with delivery ≥ 37 weeks was estimated. RESULTS: Combined screening with maternal factors and serum PAPP-A at 11-13 and 30-34 weeks and with maternal factors and serum free ß-hCG at 19-24 and 30-34 weeks improved the prediction provided by maternal factors alone for preterm PE. The detection rate, at a 10% false-positive rate, for preterm PE by screening with maternal factors was about 45% which improved to 51% and 53% by combined screening with PAPP-A at 11-13 weeks and 30-34 weeks, respectively, and 55% and 54% by combined screening with free ß-hCG at 19-24 weeks and 30-34 weeks, respectively. Measurement of serum PAPP-A and free ß-hCG was not useful in the prediction of term PE. CONCLUSIONS: Measurement of serum PAPP-A and free ß-hCG could improve the prediction of preterm PE provided by maternal characteristics and medical history alone. Copyright © 2016 ISUOG. Published by John Wiley & Sons Ltd.

Mild primary hyperparathyroidism as defined in the Italian Society of Endocrinology's Consensus Statement: prevalence and clinical features.

BACKGROUND: Mild primary hyperparathyroidism (PHPT) was recently clearly defined for the first time. Our study was thus aimed to pinpoint proportion and clinical characteristics of this kind of patients. DESIGN AND PATIENTS: We retrospectively evaluated our series of 360 consecutive patients with PHPT, selecting those with all features allowing a correct classification (serum total and ionized calcium, phosphate, creatinine, PTH, 25OHD, urinary calcium, renal and neck ultrasound, MIBI scintiscan, and DEXA at lumbar spine, femoral neck, and distal third of radius). Patients were defined asymptomatic (aPHPT) when bone or kidney was not involved and no hypercalcemic symptom occurred; mild PHPT was defined as aPHPT not meeting updated surgical criteria. RESULTS: Seventy-five patients among 172 classified as aPHPT had all available data required for surgical evaluation and could be evaluated. Sixty/75 met surgical criteria and the remaining 15 were classified as mild. Mild PHPT patients had lower total and ionized calcium, urinary calcium, and PTH levels than aPHPT meeting surgical criteria, while vitamin D levels and BMD were similar. CONCLUSIONS: Mild PHPT strictly defined according to the last consensus represents a small subgroup with a less active form of the disease.

Membrane fluctuations of human red blood cells investigated by the current signal noise in scanning ion conductance microscopy.

Human red blood cells show submicron membrane fluctuations (CMFs) that have been mainly studied with optical microscopies. Although the functional role of this phenomenon is still uncertain, the amplitude of membrane fluctuations is considered as an indicator of mechanical resilience to the stress encountered in the capillary beds. We investigate here the membrane fluctuations in red blood cells using the scanning ion conductance microscopy (SICM), a scanning probe technique that avoids the probe-sample contact. The ion current noise was recorded at a fixed distance from the cell and converted in terms of membrane fluctuation amplitude using as a converting factor the slope of the current-distance curve. We found that CMF amplitude was irreversibly reduced by membrane cross-link. Both whole cell and local increase of membrane tension induced a reduction of CMF amplitude. As for the biochemical regulation of membrane dynamics, we observed that the activation of the noradrenergic transduction pathway, via ß-receptors, increased the CMF amplitude. We conclude that the CMFs recorded by SICM and those optically recorded on red blood cells share the main features. In addition SICM provides high spatial and temporal resolution as well as the possibility to apply through the glass pipette acting as probe chemical or physical stimuli to the membrane area where the CMFs are recorded.

[Memory, fluency, and orientation: a five-minute screening test for cognitive decline]. / Memoria, fluidez y orientación: prueba de cribado de deterioro cognitivo en 5 minutos.

BACKGROUND: The prevalence of cognitive impairment (CI) will double in the next 20 years, making early detection a key priority. OBJECTIVES: Validation of a 5-minute CI screening test. METHODS: Adults aged 60 and older were recruited from memory clinics and the community at large in the Santiago, Chile metropolitan area. Based on clinical examination they were categorised as No CI (NCI), Mild CI (MCI) and dementia sufferers (DS). We measured the validity of a new test, MEFO, evaluating memory (5 points), phonetic verbal fluency (2 points) and orientation (6 points) by comparing its results with those from the MMSE. RESULTS: We evaluated 214 subjects, comprising 49 with dementia, 47 with MCI, and 118 with no CI. The MEFO differentiated between all 3 groups whereas the MMSE did not discriminate between the MCI and NCI groups. The area under the ROC curve (AUC) for the MEFO distinguishing NCI subjects from dementia sufferers was 0.97; for NCI vs CI (dementia+MCI), 0.89; and for NCI vs MCI, 0.80. On the MMSE these values were 0.95, 0.84, and 0.73, respectively. A cut-off score of 6/7 on the MEFO identified dementia sufferers with a sensitivity of 86% and a specificity of 96%. A cut-off score of 8/9 distinguished CI from NCI subjects with a sensitivity of 83% and a specificity of 75%. CONCLUSIONS: The MEFO is a valid and reliable test for discriminating between dementia and CI sufferers and subjects with no CI. Its validity is similar to that the MMSE under these conditions, but it is more effective for identifying subjects with MCI and its administration time is shorter.

Bergapten induces ER depletion in breast cancer cells through SMAD4-mediated ubiquitination.

ERα function is crucial for the development of normal mammary gland as well as in the process of progression of breast cancer cells. Signals that target receptor levels contribute to regulate estrogens effects in the cells. An intricate cross-regulation has been documented between ERα and TGF-ß down-stream molecules: SMAD2, SMAD3, and SMAD4, that can bind ERα and regulate their signaling. Thus, identification of natural anticancer drugs able to influence the latter molecule might provide alternative choices for breast cancer treatment. Taking into account our previous published data we wanted to study the effect of 5-Methoxypsoralen (bergapten) on ERα and on TGF-ß pathway. We reported that bergapten, a coumarin containing compound, effectively depletes ERα in MCF-7 breast cancer sensitive cells and in tamoxifen-resistant clone. The decrease of ERα protein after bergapten treatment results from the ubiquitine-proteasome pathway as demonstrated by the use of MG-132. IP experiments with ER antibody, demonstrated that the protein has physical interaction with SMAD4 and poly-ubiquitine and the amount of ubiquitinated receptor, linked to SMAD4, is greater under bergapten. The crucial role played by SMAD4, in this process, emerges from the observation that in breast cancer cells, silencing of SMAD4, resulted in increased expression of endogenous ERα in both control and bergapten-treated cells, compared to wild- type cells. The same results were confirmed in siRNA TGF-ß RII cells. The results suggest a novel negative regulation of ERα by TGF-ß/SMAD4 in breast cancer cells and indicate that the SMAD4 protein is involved in the degradation of ERα induced by bergapten. We propose that bergapten may efficiently act as a natural antitumoral agent, able to deplete ERα from breast cancer tamoxifen-sensitive and resistant cells, thereby retraining the effect of membrane signals targeting ERα and in such way its mitogenic potentiality.

Prevalence and characteristics of metabolic syndrome in primary hyperparathyroidism.

AIMS: Primary hyperparathyroidism (pHPT) is characterized by an increased frequency of glucose tolerance abnormalities associated with insulin resistance. Few studies evaluated the prevalence of metabolic syndrome (MetS) in pHPT and whether there are differences between asymptomatic pHPT patients and symptomatic ones. Thus, we sought to investigate the prevalence of MetS in pHPT patients in comparison to the prevalence of MetS in Italian population. SUBJECTS AND METHODS: We conducted a retrospective chart review of 294 pHPT patients, of these 154 [age (mean ± SD) 58.7 ± 13.3 yr, body mass index 25.6 ± 4.8 kg/m(2); serum calcium (11.3 ± 1.2 mg/dl) 2.8 ± 0.3 mmol/l; PTH 234.8 ± 224.3 ng/l] met the inclusion criteria. A modified National Cholesterol Educational Program (NCEP)/Adult Treatment Panel III (ATP III) definition of the MetS was used. Prevalence of MetS was compared with that reported for the Italian population (Progetto Cuore Study). RESULTS: The prevalence of the MetS (34/154, 22.1%) was similar to that reported in the general Italian population. Asymptomatic pHPT patients were older (62.1 ± 12.7 vs 56.4 ± 13.2 yr, p<0.008) and showed higher prevalence of MetS than symptomatic ones (30.2% vs 16.5%, p<0.045). Moreover the prevalence of nephrolitiasis or overt bone disease was not different between patients MetS+pHPT compared to MetS-pHPT, whereas femoral bone mineral density (BMD) was higher in MetS+pHPT (p<0.003). In the logistic regression model age and femoral BMD were independent predictors of MetS. CONCLUSIONS: The prevalence of MetS in pHPT is not increased in comparison to the general population, thus, its diagnosis is not an appropriate tool to identify the additional cardiovascular risk related to pHPT. Difference in age affects the increased prevalence of MetS in asymptomatic pHPT patients.

Interplay between serum osteocalcin and insulin sensitivity in primary hyperparathyroidism.

Osteocalcin (OC) has been proposed as a regulator of insulin sensitivity in both humans and other animals. Primary hyperparathyroidism (PHPT) is characterized by high OC levels and insulin resistance. The aim of this study was to evaluate whether in PHPT the link between OC levels and blood markers of insulin resistance was maintained. In a consecutive series of 219 adult PHPT patients, serum OC as well as fasting insulin and glucose levels were measured. Insulin sensitivity was estimated by homeostatic model assessment (HOMA2-S%). The same parameters were evaluated in a subgroup of 45 patients after parathyroidectomy (PTX). PHPT patients were characterized by markedly high OC levels. After subdividing them according to glucose tolerance, it was found that OC was similar in subjects with normal glucose tolerance (NGT) and impaired glucose tolerance (IGT), while diabetic subjects had lower serum OC than those with NGT (P < 0.02) or IGT (P < 0.04). OC was negatively associated with fasting glucose and positively associated with HOMA2-S%. OC independently predicted HOMA2-S% in a multivariate analysis. In the subgroup of surgically cured PHPT patients, OC levels significantly decreased after PTX, while HOMA2-S% did not change. Our findings indicate that in PHPT there is a positive relationship between OC and glucose metabolism, OC being one of the predictors of insulin sensitivity. However, data in surgically cured patients, showing OC normalization in spite of unchanged HOMA2-S%, suggest that OC does not likely play a major role in affecting insulin sensitivity in PHPT.

Chondrodermatitis nodularis chronica helicis and photodynamic therapy: a new therapeutic option?

Chondrodermatitis nodularis chronica helicis (CNCH) is a fairly frequent disorder of unknown etiology. Although the elective therapy is surgery, local application of topical steroids, antibiotic ointments, intralesional injection of collagen, cryotherapy, curettage followed by diathermy, and CO(2) laser treatment have also been proposed. The aim of the study was to test the utility of photodynamic therapy (PDT) for CNCH. Two patients with painful CNCH underwent PDT with a 635 nm light source for 20 minutes (70 J/cm(2) ) after application of cream containing 20% 5-aminolevulinic acid (5-ALA) and occlusion for 3 hours. The lesions decreased considerably in size and pain ceased within a few weeks. The results suggest that this method can be useful for treating CNCH, especially in patients with contraindications for surgery.

Myo-inositol products in polycystic ovary syndrome (PCOS) treatment: quality, labeling accuracy, and cost comparison.

OBJECTIVES: To evaluate the labeling accuracy of four myo-inositol products, designed for polycystic ovary syndrome (PCOS) treatment, available on the italian market and to perform a cost comparison based on myo-inositol content in milligrams for products analyzed. MATERIALS AND METHODS: Four (4) myo-inositol products (3 sachet and 1 tablet formulations) were dissolved using water, and each sample was analyzed for myo-inositol content using a high-performance liquid chromatography (HPLC) method with index refraction detector. The amount of myo-inositol per purchased product was then divided into its purchase price in order to make cost comparisons between the products based on a 2 and 4 g/day dose. RESULTS: A significant difference in the myo-inositol content, compared with the labeling was found for the products. Only 1 product contained more than 95% of the myo-inositol content claimed on the label, and there was a product with less than 75% of the labeling amount. Based on a 2-g myo-inositol per day dose, the cost of a 30-day supply ranged from Euro 20,77 and Euro 71,86, after correction by actual amount of myo-inositol. CONCLUSION: There is a lack of conformity between declared and actual amount of myo-inositol among the products tested and the majority of the products contained less than 95% of labeled amounts. There should be a better control in the manufacturing process in order to ensure more quality and accuracy. Nowadays consumers cannot trust myo-inositol product labels to represent the product's content accurately or that product pricing is a reflection of myo-inositol content.

Nutrients and infertility: an alternative perspective.

BACKGROUND: Different gonadotrophin preparations or different protocols of ovulation induction are powerless to determine a significantly increase in oocyte quality in the majority of aged patients or in patients with repeated failed in vitro fertilization-embryo transfer (IVF-ET) cycles. INFORMATION SOURCES: Papers published on journal focused on nutraceutical and human reproduction. EVIDENCES: It is questionable if various molecules that are positively associated with higher oocyte competence, higher fertilization rate and embryo development could be supplemented to infertile patients with the aim to partly reduce the frequency of unsuccessful IVF. PERSPECTIVES: Aim of this short review is mainly to focus the attention on potentially positive effects in female fertility of few, well established substances, that could be suggested as a dietary supplement.

Congenital lung malformations: a novel application for lung ultrasound?

Congenital lung malformations (CLMs) include a group of different disorders. With widespread use of antenatal ultrasonography (aUS) and increased use of pre-natal magnetic resonance imaging (MRI), CLMs are increasingly detected, nevertheless the best postnatal imaging approach is not yet well defined: newborns usually undergo several chest X-rays and eventually computed tomography to confirm the diagnosis. In this case series, we show lung ultrasound features of three different cases of congenital lung malformations, describing prenatal and postnatal images comparing different imaging techniques.

DR (Ia-like) antigens on human melanoma cells. Serological detection and immunochemical characterization.

11 cultured human melanoma cell lines were tested for the expression of DR antigens by using specific allo- and xenoantisera in an indirect rosette microassay. Four of these melanoma cell lines expressed DR antigens, but in lower amounts than expressed on cultured human B-lymphoid cells. Rabbits injected with the DR-positive melanoma cells produced antibodies that were serologically and immunochemically reactive with B-cell-derived DR antigens. Immunochemical studies indicate that melanoma cell-derived DR antigens have a two-chain structure with 34,000 and 27,000 mol wt components. The melanoma cell-derived DR beta-chain at 27,000 mol wt is slightly smaller than that of the Victor cell DR beta-chain whose mol wt is 29,000.