BACKGROUND: Beyond environmental, socioeconomic, or nutritional causes, genetic etiologies are still being uncovered as contributors to deviations from average pubertal timing determined by population studies. Central precocious puberty is a pubertal disorder that is characterized by reaching certain pubertal markers before the appropriate sex-specific age the markers should be reached at. Although there are many contributing components to this disorder, familial and twin studies strengthen the argument that genetics play a crucial role in understanding the mechanisms behind these pubertal timing deviations. Currently, there are five known genetic variants that have been linked to CPP: Makorin Ring Finger Protein 3 (MKRN3), Delta-like noncanonical Notch ligand 1 (DLK1), Prokineticin receptor 2 (PROKR2), Kisspeptin (KISS1), and its receptor (KISS1R). Any other genetic etiologies of CPP remain to be elucidated, especially when contrasted to the more than 50 genes identified with another pubertal timing disorder, hypogonadotropic hypogonadism (e.g. delayed puberty). OBJECTIVE: CPP was previously described to be mostly ‘idiopathic’ unless a pathologic organic cause was found to be contributing to the disorder. With the growing familial studies that aid in identifying the known variants as contributors to a patients’ CPP, one must wonder whether there are any unknown variants in other suspected genes that could also contribute to CPP. It is hypothesized that children with a previous diagnosis of ‘idiopathic’ CPP could potentially have an uncovered monogenic etiology for their pubertal disorder. With an emphasis on family history, this study aims to understand the inheritance patterns that influence any genetic etiologies for the improvement of familial screening, as well as to integrate any findings to a patient’s medical record. METHODS: The recruitment of participants was based on Boston Children’s Hospital Endocrine Clinic and satellite locations. Participants that were selected for recruitment met the inclusion criteria of a physician confirmed CPP diagnosis and would subsequently perform a buccal swab collection for whole-exome sequencing analysis (WES). The WES data would then be uploaded to AI genomic platform called Emedgene that allowed for further analysis into variant information including allele frequency, related diseases, and familial zygosity patterns. RESULTS: There are 22 returned WES data for analysis from patients. A frameshift (p.Met268ValfsTer23) and missense mutation (p.Arg345Cys ) affecting the gene MKRN3 were identified in two female patients in the cohort respectively. The protein areas the mutations were found to be affecting are causative to MKRN3 dysfunction based on previous studies. CONCLUSION: Previously described cases of idiopathic CPP are now being reported as cases with known or novel monogenic causes. With the growing amount of genetic variants that cause CPP being uncovered, in addition to the inheritance imprinting nature of these genes, there is an emphasis on expanding familial genetic studies to uncover the enigma behind the mechanism of pubertal regulation. Finally, the addition of clinical confirmation of genetic variants allows for research to become a part of a patient’s medical record, thus further innovating pediatric precision medicine.

As numerous bodies of research have characterized the adipocytokine adiponectin (ACRP30) as an anti-diabetic compound, this work serves as an analysis to partly elucidate the potential usage of supplementary adiponectin as a therapeutic compound in the treatment of type II diabetes mellitus. In this work, a meta-analysis was conducted to centralize large amounts of data on adiponectin’s insulin sensitizing characteristics in conjunction with like-minded studies utilizing metformin treatment as the diabetic standard of care. Upon comparison of fasting glucose, fasting insulin, and glucose tolerance test area under the curve reduction percentages, it is quite clear that ACRP30 has significant insulin enhancing properties in mouse models rivaling SOC metformin. Yet, inconsistent dosing comparisons in this analysis, the unknowns of chronic hyperadiponectinemia in humans, as well as variations in age of human diabetic patients warrant further standardized research to suggest that adiponectin supplementation can rival that of modern-day standard of care for type II diabetes mellitus.

The primary determinants of tissue glucocorticoid action are glucocorticoid receptor (GR) density and intracellular levels of ligand, the latter determined both by activity of the hypothalamic-pituitary-adrenal (HPA) axis and cellular activity of 11beta- hydroxysteroid dehydrogenase (11beta-HSD) enzymes that interconvert active 11- hydroxy (corticosterone, cortisol) and inactive 11-keto (11-dehydrocorticosterone, cortisone) glucocorticoids. Here, the contribution of GR density and ligand levels in determining body composition and metabolic phenotype have been investigated in mice and in humans. Genetic evidence in humans implicates variations in the GR gene in the regulation of the HPA axis as well as the control of body fat distribution, metabolic parameters and blood pressure. Although GR deficient mouse models have been previously generated (with homozygous nulls dying at birth), the effects of altered GR density upon fat distribution and blood pressure have not been described. This study addresses the relationship between GR density and metabolic parameters, including body fat distribution, insulin resistance and hypertension. A novel line of mice harbouring a null mutation in the GR gene (GR+/-) was generated from an ES cell line in which a beta-galactosidase-neomycin phosphotransferase (beta geo) reporter cassette was fused with GR. The resulting fusion protein lacks part of the DNA binding domain and the entire ligand binding domain and is transcriptionally inactive. In addition, the beta-galactosidase enzyme activity “reports” activity of the GR gene promoter. GR-/- mice are present in a normal Mendelian ratio before birth. Intriguingly, 1 (of 36/146 expected if null allele not lethal) survived to adulthood suggesting this might be a hypomorphic rather than a null allele. Heterozygous 15 (GR+/-) mice showed 40-45% reductions in GR mRNA levels in the hippocampus, paraventricular nucleus of the hypothalamus, pituitary gland and adipose tissue, 30% in liver, 56% in muscle and 67% in adrenals. X-gal staining of GR+/- brain sections showed that GR-beta gal is present throughout, mirroring GR mRNA expression. Adult GR+/- mice had larger adrenals, higher evening plasma corticosterone levels and greater corticosterone responses following 10 minute restraint suggesting a hyperactive HPA axis. Compared to GR+/+ littermates, GR+/- mice had similar body weight gain on normal chow or high fat diet, with unaltered fat depot (inguinal, epididymal, mesenteric) weights and similar glucose and insulin tolerance. However, GR+/ - mice had higher (10%) systolic blood pressure, associated with activation of the renin-angiotensin system. Thus GR haploinsufficiency in mice causes increased blood pressure and accords with data associating GR polymorphisms with hypertension in humans. The role of altered GC sensitivity was also investigated in a mouse model of HPA axis hypoactivity pro-opiomelanocortin null (POMC) mice. POMC-null mice are obese due to central melanocortin deficiency. In contrast to most rodent models of obesity, POMC-null mice are also glucocorticoid deficient due to ACTH deficiency. Previous data have shown that glucocorticoid replacement in POMC-null mice exaggerated hyperphagia, obesity and insulin resistance and caused hypertension. Here, the contribution of peripheral glucocorticoid sensitivity was investigated. POMC-null mice have increased liver and retroperitoneal fat GR mRNA levels but, specifically in adipose tissue, decreased levels of mRNA encoding 11beta-HSD1, a reductase which regenerates active glucocorticoids, thus amplifying their action.

No description available.

No description available.

a2u-globulin and albumin are secretory proteins synthesised in rat liver parenchymal cells and appear as major components in urine and serum, respectively. The syntheses of both proteins are under complex hormonal control. Sex-hormones appear to be particularly important in regulating a2u-globulin synthesis since a2u-globulin is normally only found in adult male rats. The intracellular site and mode of a2u-globulin synthesis were studied and a hormone-responsive hepatocyte in vitro system for studying the effects of hormones on protein synthesis was developed. Newly synthesised a2u-globulin and albumin were recognised by immunoprecipitation and SDS-polyacrylamide gel analysis of radiolabelled products. Specific antibodies were raised in rabbits to the purified proteins. Rat hepatic polysomes were isolated and used to direct translation in a polysome-free reticulocyte cell-free system. Identification of the in vitro products showed that a2u-globulin and albumin were synthesised exclusively on the membrane-bound polysomes. a2u-globulin-mRNA and albumin-mRNA were identified in the poly(A)- and non poly(A)-containing RNA fractions isolated from total hepatic RNA preparations, respectively. The molecular weights of [35S]-labelled a2u-globulin from RNA or polysome directed in vitro systems and [3H]-labelled a2u-globulin secreted from hepatocytes were compared by co-electrophoresis of the appropriate immuno-precipitates on 15 - 18% SDS polyacrylamide gels. The slight, although consistent, higher molecular weight found for the in vitro product suggests that a2u-globulin, like other secretory proteins, may involve a precursor. Rat hepatocyte cell suspensions prepared by collagenase treatment of perfused livers incorporated radiolabelled amino acids linearly into proteins for at least seven hours. The relative rates of albumin synthesis were similar in hepatocytes isolated from males, females, castrated males and ovariectomised females. a2u-globulin synthesis was found only in cells from males, and at reduced rates in those from castrated males and androgen-treated ovariectomised females. 5a-dihydrotestosterone failed to stimulate a2u-globulin, albumin and total protein synthesis in hepatocytes from castrated males and ovariectomised females but specifically doubled the rate of a2u-globulin synthesis in cells from androgen-pretreated, ovariectomised females. Although a2u-globulin synthesis rates could be stimulated or inhibited in hepatocytes from males by oestradiol-17B, depending on concentrations, these effects were non-specific. Inhibitor studies suggested the involvement of microtubules in the secretion of a2u-globulin and albumin from hepatocytes.

Descriptions are given of the cerebral ganglionic complex and retro cerebral system of neoimaginal Chironomus riparius reared at 20°C. Changes resulting from differences in age, rearing temperature and reproductive state are also presented. The anatomy of the cerebral ganglia, including the organisation of neuropile masses, of male neoimagines, and the fine structure of the neural sheath, cortical layer and neuropile is described for the first time in an adult chironomid. Two types of perineurial cells and three types of glial cells are distinguished on ultra structural criteria. A classification of five non-neurosecretory neuron types, based upon size, nuclear to cytoplasmic proportions and appearance, is given. Four paired groups of neurosecretory neurons were found in the brain and two paired groups in the suboesophageal ganglion. Five neurosecretory cell types are classified on the basis of their ultrastructural characteristics. The axon pathways of the cerebral neurosecretory cells and the elements of the retrocerebral system, including the paired corpora cardiaca, corpora allata, peritracheal tissues and "glandes post-cerebrales ant erieures", are described. The nervous and endocrine structures of the neoi maginal male are compared to those of the female. Age related changes occurring within the brain and endocrine system are described in relation to fat body depletion, behavioural activity and appearance of the dorsal longitudinal flight muscles. The effect that differences in rearing temperature have upon the longevity, size and weight of the adult is presented. The influence of rearing temperature upon the ultrastructure of the brain is described for the first time in an insect. Evidence suggesting an association between the activity of the cerebral neurosecretory cells and female reproduction is given. In the light of the results functional roles for several elements within the endocrine system are postulated. The possible significance that the observed structural changes in the nervous system have upon its functioning is discussed.

<p> Infusion of 17&beta;-estradiol into the arcuate nucleus of the hypothalamus (ARH) 47.5 hours after estrogen benzoate (EB) priming rapidly facilitates sexual receptivity (lordosis) via G protein-coupled estrogen receptor 1 (GPER) that deactivates &micro;-opioid receptors (MOP) in the medial pre-optic nucleus of the hypothalamus (MPN). Initial estradiol inhibits lordosis via activating an ARH neurocircuit that activates MPN MOP, and simultaneously upregulates orphanin FQ/ nociception (OFQ/N), its cognate receptor (ORL-1), and progesterone receptor (PR) in the ARH. Subsequent ORL-1 activation 48 hours post-EB deactivates MPN MOP to facilitate lordosis. Thus, I hypothesized that GPER directly regulates OFQ/N neurons and tested whether EB increased coexpression of GPER and OFQ/N in ARH neurons. EB significantly increased the number of GPER OFQ/N expressing neurons. Antiestrogens, tamoxifen (TAM) and ICI 182,780 (ICI), are treatments for some estrogen responsive tumors, but sometimes exacerbate tumor proliferation via GPER. I hypothesized that TAM and ICI activate ARH GPER and facilitate lordosis via deactivation of MPN MOP. In EB-primed rats, ARH infusion of either TAM or ICI facilitated lordosis and deactivated MPN MOP. GPER antagonist, G15, blocked these results. Thus, TAM and ICI rapidly activate ARH GPER neurons that express OFQ/N to facilitate sexual receptivity.</p>

Physicians and scientists have suspected that the testes secrete a substance into the body that causes male secondary sexual characteristics for hundreds of years. However, testosterone was not synthesized until 1935 and it was not until the 1940's when scientists could accurately measure the amount of this hormone in the blood. Since then, scientists have been able to make correlations between the levels of testosterone in the body and men's health Scientists have long observed higher levels of testosterone to be associated with an increase in levels of Hematocrit (Hct). As a result, Testosterone Replacement Therapy (TRT) has been used to treat anemia associated with chronic diseases. In recent years, prescription sales for testosterone have sky rocketed due to new clinical uses such as androgen deficiency in older men. In fact, the rate of prescription for testosterone products has increased by over 170% in the previous five years. Long-term data shows that the level of testosterone in the male body begins to decrease at about the age of 30. As the life expectancy of the general population continues to increase, TRT may be a viable option for older men with low testosterone to increase the quality and duration of life. However, an increase in Hct continues to be a major side effect of TRT. New research is beginning to make clear the mechanism by which testosterone affects erythropoeisis. New research suggests TRT suppresses hepcidin and leads to an increase in the rate of iron (Fe) retention in red blood cells (RBCs). Inter-individual differences in the pharmacogenetic effects of TRT have been observed. In the future TRT could be genetically tailored based on the individuals DNA. In this case, the optimal dose of testosterone can be given to maximize benefits and reduce side effects. Here, the risks and benefits associated with TRT and a review of the updated Clinical Guidelines for its use will be presented. The effects of TRT on erythropoeisis will be investigated via a review of the literature. The main objective of this review is to provide a general understanding of TRT and a major side effect of its use, excessive erythropoeisis.

No description available.