Differential expression of FMR1, FXR1 and FXR2 proteins in human brain and testis

Lack of expression of the fragile X mental retardation protein (FMRP) results in mental retardation and macroorchidism, seen as the major pathological symptoms in fragile X patients. FMRP is a cytoplasmic RNA- binding protein which cosediments with the 60S ribosomal subunit. Recently, two proteins homologous to FMRP were discovered: FXR1 and FXR2. These novel proteins interact with FMRP and with each other and they are also associated with the 60S ribosomal subunit. Here, we studied the expression pattern of the three proteins in brain and testis by immunohistochemistry. In adult brain, FMR1, FXR1 and FXR2 proteins are coexpressed in the cytoplasm of specific differentiated neurons only. However, we observed a different expression pattern in fetal brain as well as in adult and fetal testis, suggesting independent functions for the three proteins in those tissues during embryonic development and adult life.      

The Portuguese validation of the International Consultation on Incontinence Questionnaire—Vaginal Symptoms (ICIQ-VS) for Brazilian women with pelvic organ prolapse

The aim of this study is to validate the International Consultation on Incontinence Questionnaire-Vaginal Symptoms (ICIQ-VS) in Portuguese. Two hundred four women (108 symptomatic, 94 asymptomatic, and two with no data) with mean age of 55.4 years received a Portuguese version of the ICIQ-VS. Clinical data and pelvic organ prolapse quantification index (POP-Q) were obtained. Retest was performed 3 weeks later. Responsiveness was assessed after 20 weeks of postsurgical follow-up. Overall, most patients presented POP-Q > 2. ICIQ-VS demonstrated good psychometric properties (validity, reliability and responsiveness). The test-retest reliability was moderate to excellent for all questions. The construct validation distinguished differences in ICIQ-VS scores between symptomatic (ICIQ-VS5a > 0) and asymptomatic (ICIQ-VS5a = 0) women. ICIQ-VS was highly responsive to surgical treatment and discriminated between levels of change in the vaginal symptoms score, sexual matters score, quality-of-life score, and POP-Q. The Portuguese version of ICIQ-VS was successfully validated.     

Growth hormone secretagogue (GHS) analogue, hexarelin stimulates GH from peripheral lymphocytes.

The role of growth hormone releasing hormone (GHRH) and growth hormone releasing peptide-6 (GHRP-6) analogue hexarelin was investigated in the regulation of GH production from lymphocytes. Porcine and bovine blood mononuclear cells were separated using density gradient centrifugation method by layering the whole blood or buffy coat cells on lymphodex. Cells were incubated for 3 or 5 days with or without phytohemagglutinin (PHA-M), GHRH, GHRP-6 analogue hexarelin, somatostatin or GHRH + hexarelin. Growth hormone was fractionated from supernatants by gel chromatography and further concentrated by lyophilization at - 20 degrees C. A nearly two fold increase in basal secretion of GH (porcine: 3.5 +/- 0.1 ng/ml, bovine: 3.2 +/- 0.2 ng/ml) was achieved by GHRH and hexarelin at concentrations of 0.1, 1.0, 10 and 100 nM in both porcine and bovine cells. Lymphocytic GH release was also stimulated in response to PHA-M (10 micro g/well). Neither a dose dependent nor a synergistic nor an additive effect was apparent on GH secretion from lymphocytes. GHRH stimulated lymphocytic GH secretion, whereas, somatostatin had no effect. This study reports for the first time that hexarelin stimulates the secretion of GH from peripheral lymphocytes.     

Role of androgens in proliferation and differentiation of mouse mammary epithelial cell line HC11

Androgens have been found in mammary epithelium and in milk throughout the cycle of the mammary gland in vivo. The aim of this study was to investigate the possible role of these substances in mammary epithelial growth and differentiation in the mouse HC11 cell line. Cells were stimulated with testosterone, dihydrotestosterone, androstenedione and 5alpha-androstane-3alpha,17beta-diol at concentrations ranging between 0.3 nM and 30 nM. Cyproterone acetate or flutamide, androgen receptor antagonists, (3 microM) were used to block specific androgen effects. Proliferative effects were measured by an MTT (tetrazolium blue) conversion test and [(3)H]thymidine uptake. HC11 cells were transfected with pbetacCAT, a chimeric rat beta-casein gene promoter-chloramphenicol acetyl transferase (CAT) gene construct and CAT ELISA was used to determine gene expression. RT-PCR was performed to detect androgen receptor expression. After 24, 48 and 72 h androgens significantly (P<0.05) increased proliferation. Androgen antagonists significantly (P<0.05) reduced the proliferative effects. Furthermore androgens potentiated the lactogenic effect of prolactin, insulin and dexamethasone (P<0.05). Finally, the androgen receptor gene was expressed in both proliferating and differentiated HC11 cells. These observations lead us to hypothesize an activity of this class of steroids in mammary physiology. In particular, androgens stimulate cell proliferation and beta-casein gene expression; this influence appears to be mediated by androgen receptors.     

Cortisol determination in hair and faeces from domestic cats and dogs

The present study explored the feasibility of a hair cortisol assay in domestic cats (Felis silvestris catus) and dogs (Canis familiaris) as a valid and reliable alternative to existing non-invasive techniques for monitoring the hypothalamic-pituitary-adrenal (HPA) axis activity. To this aim, 56 new hair growth samples and 870 faecal samples from 27 domestic cats and 29 domestic dogs were collected and cortisol content was assessed. A significant positive association was observed in both species between the concentrations of cortisol determined in hair and faeces. This finding is discussed in the light of the existing knowledge of hair physiology and in the perspective of its application to studies on chronic stress.     

Protracted neonatal hypertrypsinogenaemia, normal sweat chloride, and cystic fibrosis.

The cystic fibrosis (CF) clinical spectrum has greatly expanded in the past few years, including atypical forms with low sweat chloride concentrations. Two cases are presented which suggest that children detected by neonatal CF screening whose trypsinogen concentrations are still raised by the second month of age could, despite a negative sweat test, be affected by an atypical CF with fully expressed pulmonary involvement.     

Pancreatic phenotype in infants with cystic fibrosis identified by mutation screening.

OBJECTIVE: To determine the pancreatic phenotype of infants with cystic fibrosis (CF) diagnosed in the first week of life by a combined immunoreactive trypsin/mutation screening program. DESIGN: A prospective evaluation of pancreatic function in infants with CF at the time of neonatal diagnosis and up to the age of 12. SETTING: Two different centres (Verona, Italy and Westmead, Australia) to enable comparison of results between two regions where <60% or > or =90% of patients, respectively, have at least one single DeltaF508 a mutation. Patients: 315 children with CF including 149 at Verona and 166 at Westmead. INTERVENTIONS: Fat balance studies over 3-5 days and pancreatic stimulation tests with main outcome measures being faecal fat or pancreatic colipase secretion. Patients with malabsorption are pancreatic insufficient (PI) or with normal absorption and pancreatic sufficient (PS). RESULTS: 34 infants (23%) at Verona and 46 (28%) at Westmead were PS at diagnosis. 15% of those with two class I, II or III "severe" mutations and 26/28 (93%) of those with class IV or V mutations were PS at this early age. Of the 80 infants with PS, 20 became PI before the age of 12. All 20 had two severe mutations. CONCLUSION: Neonatal mutational screening programs for CF are less likely to detect PS patients with non-DeltaF508 mutations. Of PS patients who are detected, those with two severe class I, II or III mutations are at particularly high risk of becoming PI during early childhood.     

Conservative management of malignant gastric outlet obstruction syndrome-evidence based evaluation of endoscopic ultrasound-guided gastroentero-anastomosis

Gastric outlet obstruction (GOO) is a clinical syndrome characterized by postprandial vomiting, abdominal pain, bloating and, in advanced cases, by weight loss secondary to inadequate oral intake. This clinical entity may be caused by mechanical obstruction, either benign or malignant, or by motility disorders. In this review we will focus on malignant GOO and on its endoscopic ultrasound (EUS)-guided palliative treatment. The most frequent malignant causes of this syndrome are gastric and locally advanced pancreatic carcinomas; other causes include duodenal or ampullary neoplasms, gastric lymphomas, retroperitoneal lymphadenopathies and, more infrequently, gallbladder and bile duct cancers. Surgery represents the treatment of choice when radical and curative resection is potentially feasible; if the malignant cause is not likely to be completely resected, palliative treatments should be proposed. Palliative treatments for malignant GOO are primarily based on surgical gastro-jejunostomy and endoscopic placement of an enteral self-expanding metal stent. Both treatments are effective; however, endoscopic stent placement is less invasive and it is associated with good short-term results, while surgery provides longer-lasting effects with a lower frequency of reintervention. In the last few years, EUS-guided gastroenterostomy (GE) has been proposed as palliative treatment for malignant GOO. This novel technique consists of the creation of an anastomosis between the gastric lumen and a small bowel loop distal to the malignant obstruction, through the deployment of a lumen-apposing metal stent under EUS-view. EUS-GE has the advantage of being as minimally invasive as enteral stent placement, and of guaranteeing long-term results similar to those of surgery.     

A family of transmembrane proteins with homology to the MET-hepatocyte growth factor receptor.

In hunting for unknown genes on the human X chromosome, we identified a cDNA in Xq28 encoding a transmembrane protein (SEX) of 1871 amino acids. SEX shares significant homology with the extracellular domain of the receptors encoded by the oncogenes MET, RON, and SEA [hepatocyte growth factor (HGF) receptor family]. Further screenings of cDNA libraries identified three additional sequences closely related to SEX: these were named SEP, OCT, and NOV and were located on human chromosomes 3p, 1, and 3q, respectively. The proteins encoded by these genes contain large cytoplasmic domains characterized by a distinctive highly conserved sequence (SEX domain). Northern blot analysis revealed different expression of the SEX family of genes in fetal tissues, with SEX, OCT, and NOV predominantly expressed in brain, and SEP expressed at highest levels in kidney. In situ hybridization analysis revealed that SEX has a distinctive pattern of expression in the developing nervous system of the mouse, where it is found in postmitotic neurons from the first stages of neuronal differentiation (9.5 day postcoitus). The SEX protein (220 kDa) is glycosylated and exposed at the cell surface. Unlike the receptors of the HGF family, p220SEX, a MET-SEX chimera or a constitutively dimerized TPR-SEX does not show tyrosine kinase activity. These data define a gene family (SEX family) involved in the development of neural and epithelial tissues, which encodes putative receptors with unexpected enzymatic or binding properties.     

Progesterone does not inhibit the increase in pituitary content of luteinizing hormone after removal of estradiol in the ewe

During late gestation in the ewe, the pituitary content of LH is reduced by about 95%, presumably due to the presence of high concentrations of ovarian steroids. The aim of this study was to determine whether the pituitary content of LH in the ewe can increase after long-term administration of ovarian steroids, when only estradiol (E) is removed or if both E and progesterone (P) must be withdrawn to allow synthesis of LH to occur. Ten ovariectomized ewes were treated with implants containing E and P. After 3 weeks of treatment, the E implants were removed from 5 ewes (-E+P) and both steroid implants were removed from the remaining 5 ewes (-E-P). Five ovariectomized ewes received P implants at the beginning of the experiment and these implants were left in place for the duration of the study; 5 ovariectomized ewes served as controls (C). All animals were injected with 100 micrograms GnRH iv 3, 6 and 9 weeks after the initiation of treatment. The area under the LH-response curve was used as an indication of the pituitary content of LH. All steroid treatments markedly reduced basal levels of LH. LH levels increased only in -E-P ewes, beginning 6 weeks after initiation of the study. After 3 weeks, -E+P and -E-P ewes released less LH (P less than 0.05) in response to GnRH than did C ewes, whereas P animals did not differ from controls. LH release in response to GnRH in -E+P and -E-P groups had increased by 6 and 9 weeks and was not different from that of C ewes.(ABSTRACT TRUNCATED AT 250 WORDS)