Hormone Producing Research Articles

Immunohistomorphometric Changes of The Pituitary Gonadotropic Cells After Testosterone Application in a Rat Model of the Andropause

Abstract Andropause, the culminating phase of male ageing, is characterized by deregulation of the hypothalamic-pituitarygonadal axis and low circulating free testosterone. The aim of this study was to investigate the immunohistomorphometric characteristics of the pituitary gonadotropic i.e. follicle-stimulating hormone (FSH) and the luteinizing hormone (LH) producing cells after testosterone application in a rat model of the andropause. Middle-aged Wistar rats were divided into orchidectomized (ORX; n=8) and testosterone treated orchidectomized (ORX+T; n=8) groups. Testosterone propionate (5 mg/kg b.m./day) was administered for three weeks, while the ORX group received the vehicle alone. Immunohistochemically stained FSH and LH cells underwent morphometric and optical density-related analysis, while circulating concentrations of the sex steroids were measured by immunoassays. Serum concentrations of testosterone and estradiol were significantly (p<0.05) increased by 24 and 2.7 fold respectively, compared to the ORX group. The volume of FSH and LH cells was significantly (p<0.05) decreased by 51.3% and 56.6% respectively, in comparison with ORX rats. Relative volume density of FSH and LH cells was also significantly (p<0.05) decreased by 54.0% and 72.8% respectively, compared to the ORX group. Results related to the optical density of gonadotropic cells (reflecting their hormonal content) were in line with the morphometric findings i.e. this parameter of FSH and LH cells was significantly (p<0.05) decreased by 25.7% and 16.2% respectively, in comparison with ORX rats. Conclusion: In conclusion, applied testosterone increased the serum concentrations of sex steroids, as well as it decreased morphometric parameters and optical density of gonadotropic cells in ORX rats.

Next-generation sequencing identifies novel mitochondrial variants in pituitary adenomas

PurposeDisrupted mitochondrial functions and genetic variants of mitochondrial DNA (mtDNA) have been observed in different human neoplasms. Next-generation sequencing (NGS) can be used to detect even low heteroplasmy-level mtDNA variants. We aimed to investigate the mitochondrial genome in pituitary adenomas by NGS.MethodsWe analysed 11 growth hormone producing and 33 non-functioning [22 gonadotroph and 11 hormone immunonegative] pituitary adenomas using VariantPro™ Mitochondrion Panel on Illumina MiSeq instrument. Revised Cambridge Reference Sequence (rCRS) of the mtDNA was used as reference. Heteroplasmy was determined using a 3% cutoff.Results496 variants were identified in pituitary adenomas with overall low level of heteroplasmy (7.22%). On average, 35 variants were detected per sample. Samples harbouring the highest number of variants had the highest Ki-67 indices independently of histological subtypes. We identified eight variants (A11251G, T4216C, T16126C, C15452A, T14798C, A188G, G185A, and T16093C) with different prevalences among different histological groups. T16189C was found in 40% of non-recurrent adenomas, while it was not present in the recurrent ones. T14798C and T4216C were confirmed by Sanger sequencing in all 44 samples. 100% concordance was found between NGS and Sanger method.ConclusionsNGS is a reliable method for investigating mitochondrial genome and heteroplasmy in pituitary adenomas. Out of the 496 detected variants, 414 have not been previously reported in pituitary adenoma. The high number of mtDNA variants may contribute to adenoma genesis, and some variants (i.e., T16189C) might associate with benign behaviour.

Exploiting novel tailored immunotherapies of type 1 diabetes: Short interfering RNA delivered by cationic liposomes enables efficient down-regulation of variant PTPN22 gene in T lymphocytes

In autoimmune diseases as Type 1 diabetes, the actual treatment that provides the missing hormones is not able, however, to interrupt the underlining immunological mechanism. Importantly, novel immunotherapies are exploited to protect and rescue the remaining hormone producing cells. Among probable targets of immunotherapy, the C1858T mutation in the PTPN22 gene, which encodes for the lymphoid tyrosine phosphatase (Lyp) variant R620W, reveals an autoimmunity related pathophysiological role. Our scope was to establish new C1858T PTPN22 siRNA duplexes delivered by liposomal carriers (lipoplexes) to patients’ PBMC. Following lipoplexes treatment, CD3+ and CD3− immunotypes were efficiently transfected; cell integrity and viability were preserved. Specific target mRNA down-modulation was observed. After T cell receptor stimulation, in lipoplexes-treated PBMC Lyp function was restored by increased release of IL-2 in cultures. Results set-up the stage for ultimate trials in the treatment of autoimmunity based on the specific inhibitory targeting of C1858T PTPN22 by lipoplexes.

Morphofunctional parameters of rat somatotrophes after acute and repeated immobilization or restraint stress

It is well known that stress changes levels of pituitary hormones in the bloodstream and in the pituitary itself. However, almost nothing is known about the impact of stress on histological and stereological parameters of the growth hormone producing cells (somatotrophs-GH cells). The aim of the present study was to investigate the effect of: acute and repeated immobilization; acute and repeated restraint on histological and morphofunctional parameters of somatotrophs in adult Wistar rats. Changes in the pituitary gland volume; the volume density and volume of somatotrophs following acute and repeated immobilization (IMO, R-IMO); acute and repeated restraint (R, R-R) were evaluated using a stereological system (newCAST), while growth hormone level within pituitary was determined by Western blot. Our results demonstrated the decrease (p < 0.05) of the pituitary volume (17%, 19%) in the IMO and R groups, respectively, and the increase in the R-R group. The volume density of GH cells decreased (p < 0.05) in the R-IMO (7%), R (26%) and R-R (18%) group in comparison to the control value. The pituitary GH content was increased (p < 0.05) after the IMO (2-fold), R (2.5-fold) and R-R (2.1-fold) as compared to the control group. These results point out that acute and repeated immobilization and/or restraint lead not only to changes in GH hormone concentration, but also modify the morphological aspects of GH cells within the rat pituitary.

Pheochromocytoma and Paraganglioma in Neurofibromatosis type 1: frequent surgeries and cardiovascular crises indicate the need for screening

BackgroundPheochromocytomas and Paragangliomas (PCC/PGL) are rare endocrine tumors that are mostly benign, but often hormone producing, causing significant morbidity and mortality due to excess catecholamine secretion and cardiovascular crises. It is estimated that 30% of PCC/PGL are due to germline mutations, including Neurofibromatosis type 1 (NF1). There is little published data describing the phenotype of NF1-associated PCC/PGL and there are no established recommendations for PCC/PGL screening in NF1.MethodsWe conducted a retrospective chart review of 17 patients with NF1-associated PCC/PGL who received care at a large academic referral center between the years of 1992–2016.ResultsAverage age of diagnosis was 42 years old. Both genders were equally affected. Average tumor size was 3.9 cm. Nine patients were hypertensive; one had orthostatic hypotension; three had tachycardia; the remaining two patients had normal BP and HR. Most tumors were benign, unilateral adrenal tumors that were hormonally active. Two had metastatic disease. Six patients experienced cardiovascular crises; three of which occurred during elective surgeries for neurofibroma removal, and a fourth occurred during labor and delivery.ConclusionThese data highlight the importance of screening for PCC/PGL in NF1, especially prior to surgical procedures and pregnancy, labor and delivery as these events can trigger a cardiovascular crisis. Screening is easily accomplished with plasma or urine free fractionated metanephrine levels.

Giant pituitary adenoma: histological types, clinical features and therapeutic approaches.

Giant pituitary adenomas comprise about 6-10% of all pituitary tumors. They are mostly clinically non-functioning adenomas and occur predominantly in males. The presenting symptoms are usually secondary to compression of neighboring structures, but also due to partial or total hypopituitarism. Functioning adenomas give rise to specific symptoms of hormonal hypersecretion. The use of dopamine agonists is considered a first-line treatment in patients with giant macroprolactinomas. Somatostatin analogs can also be used as primary treatment in cases of growth hormone and thyrotropin producing giant adenomas, although remission of the disease is not achieved in the vast majority of these patients. Neurosurgical treatment, either through transsphenoidal or transcranial surgery, continues to be the treatment of choice in the majority of patients with giant pituitary adenomas. The intrinsic complexity of these tumors requires the use of different therapies in a combined or sequential way. A multimodal approach and a therapeutic strategy involving a multidisciplinary team of expert professionals form the basis of the therapeutic success in these patients.

Diagnosis and Management of Gynecomastia for Urologists.

Our aim is to review the steps of diagnosis and management of gynecomastia with a special focus on treatment of gynecomastia associated with androgen deprivation therapy for prostate cancer. Recent studies investigating tamoxifen and radiation therapy for both therapy and prophylaxis of bicalutamide-induced gynecomastia are reviewed. Gynecomastia is a common clinical problem, affecting between one and two thirds of middle-aged men. Diagnosis is typically made by history and physical exam. Common causes include chronic medical conditions and medications; however, unexplained gynecomastia should prompt laboratory work-up, followed by appropriate imaging studies to evaluate for hormone producing cancers. For patients taking bicalutamide for treatment of prostate cancer, tamoxifen or radiation therapy for gynecomastia are excellent options.

Microporous Polymer Scaffolds for the Transplantation of Embryonic Stem Cell Derived Pancreatic Progenitors to a Clinically Translatable Site for the Treatment of Type I Diabetes.

Type I diabetes mellitus, which affects an estimated 1.5 million Americans, is caused by autoimmune destruction of the pancreatic beta cells that results in the need for life-long insulin therapy. Allogeneic islet transplantation for the treatment of type I diabetes is a therapy in which donor islets are infused intrahepatically, which has led to the transient reversal of diabetes. However, therapeutic limitations of allogeneic transplantation, which include a shortage of donor islets, long-term immunosuppression, and high risk of tissue rejection, have led to the investigation of embryonic or induced pluripotent stem cells as an unlimited source of functional beta-cells. Herein, we investigate the use of microporous scaffolds for their ability to promote the engraftment of stem cell derived pancreatic progenitors and their maturation toward mono-hormonal insulin producing β-cells at a clinically translatable, extrahepatic site. Initial studies demonstrated that microporous scaffolds supported cell engraftment, and their maturation to become insulin positive; however, the number of insulin positive cells and the levels of C-peptide secretion were substantially lower than what was observed with progenitor cell transplantation into the kidney capsule. The scaffolds were subsequently modified to provide a sustained release of exendin-4, which has previously been employed to promote maturation of pancreatic progenitors in vitro and has been employed to promote engraftment of transplanted islets in the peritoneal fat. Transplantation of stem cell derived pancreatic progenitors on scaffolds releasing exendin-4 led to significantly increased C-peptide production compared to scaffolds without exendin-4, with C-peptide and blood glucose levels comparable to the kidney capsule transplantation cohort. Image analysis of insulin and glucagon producing cells indicated that monohormonal insulin producing cells were significantly greater compared to glucagon producing and polyhormonal cells in scaffolds releasing exendin-4, whereas a significantly decreased percentage of insulin-producing cells were present among hormone producing cells in scaffolds without exendin-4. Collectively, a microporous scaffold, capable of localized and sustained delivery of exendin-4, enhanced the maturation and function of pluripotent stem cell derived pancreatic progenitors that were transplanted to a clinically translatable site.

PD33-10 HUMAN INDUCED PLURIPOTENT STEM CELL-DERIVED TESTOSTERONE-PRODUCING LEYDIG CELLS AMELIORATE SERUM TESTOSTERONE LEVEL IN RATS.

You have accessJournal of UrologyStem Cell Research: Stem Cell Research I1 Apr 2018PD33-10 HUMAN INDUCED PLURIPOTENT STEM CELL-DERIVED TESTOSTERONE-PRODUCING LEYDIG CELLS AMELIORATE SERUM TESTOSTERONE LEVEL IN RATS. Takaki Ishida, Mikito Tanaka, Kenta Sumii, Keisuke Okada, Koji Chiba, Kei Matsushita, Masato Fujisawa, and Takashi Aoi Takaki IshidaTakaki Ishida More articles by this author , Mikito TanakaMikito Tanaka More articles by this author , Kenta SumiiKenta Sumii More articles by this author , Keisuke OkadaKeisuke Okada More articles by this author , Koji ChibaKoji Chiba More articles by this author , Kei MatsushitaKei Matsushita More articles by this author , Masato FujisawaMasato Fujisawa More articles by this author , and Takashi AoiTakashi Aoi More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.1565AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Recently male late-onset hypogonadism syndrome (LOH syndrome) is focused on with various symptoms due to hypogonadism. Although the results of the differentiation instructions from human induced pluripotent stem cell (human iPS cells) to many various cells, there is no report that succeed in induced Leydig cell which secret androgens. In this time we succeeded in Induced Leydig cell secrete testosterone from human iPS cells. METHODS First, we transduced NR5A1 expressing system by doxycycline addition to human iPS cell. We examined various genetic expression and hormone producing ability and evaluate the differentiation instruction from this cell under a timing of the NR5A1 forced expression and various kinds of conditions. Human iPS cells-derived testosterone-producing Leydig cells were transplanted into castrated male rats and examined for improvement in serum testosterone levels. RESULTS Leydig cell marker (INSL3 variant2) and LH receptor (LHCGR) and Leydig cell-specific enzyme (17βHSD3)and expression of various enzyme (StAR,CYP11A,3βHSD2,CYP17A) necessary for testosterone production were confirmed in RT-PCR.INSL3 and LHCGR were confirmed in Immunohistochemical staining.Testosterone production was confirmed by the hormone measurement in supernatants. And we showed that transplantation of human iPS cells-derived testosterone-producing Leydig cells to castrated rats ameliorate serum testosterone levels.At the same time, expression of adrenal cortex-specific enzyme (CYP21A2,CYP11B1,CYP11B2) necessary for cortisol and aldosterone production were confirmed in RT-PCR. The cluster of Leydig cells include the adrenal cortex cells too. CONCLUSIONS We succeeded in Leydig cell instruction from human iPS cells. At the same time to adrenocortical cell is induced, and we will examine the specific differentiation instruction conditions to Leydig cell in future. Transplantation of human iPS cells-derived testosterone-producing Leydig cells to castrated rats ameliorate serum testosterone levels. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e654 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Takaki Ishida More articles by this author Mikito Tanaka More articles by this author Kenta Sumii More articles by this author Keisuke Okada More articles by this author Koji Chiba More articles by this author Kei Matsushita More articles by this author Masato Fujisawa More articles by this author Takashi Aoi More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

The Morpho-Functional Parameters of Rat Pituitary Hormone Producing Cells After Genistein Treatment

Abstract Phytoestrogens are a diverse group of steroid–like compounds that occur naturally in many plants. There are various types of phytoestrogens, including the best-researched isoflavones which are commonly found in soy. The consumption of soy products has many health benefits, including protection against breast cancer, prostate cancer, menopausal symptoms, heart disease and osteoporosis. In contrast, use of hormonally active compounds-isoflavones may unfortunately interfere with the endocrine system and can have far-reaching consequences. Genistein, the most abundant soy-bean derived isoflavone, possesses a ring system similar to estrogens and acts through an estrogen receptor (ER)-mediated mechanism, by increasing or decreasing the transcription of ER-dependent target genes. Also, genistein can act on cells through ER non-dependent mechanisms, such as tyrosine kinase inhibitor. The neuroendocrine systems are responsible for the control of homeostatic processes in the body, including reproduction, growth, metabolism and energy balance, and stress responsiveness. It is well known, that estrogen is important for development of the neuroendocrine system in both sexes. At the pituitary level, estrogen is known to affect the regulation of all hormone producing (HP) cells, by direct and/or indirect mechanisms. Due to structural and functional resemblance to estrogen, the question may arise of whether and how genistein affects the morphofunctional features of pituitary HP cells. This review deals with the consequences of genistein’s effects on morphological, stereological and hormonal features of HP cells within the anterior pituitary gland. Transparency on this issue is needed because isoflavones are presently highly consumed. Inter alia, genistein as well as other isoflavones, are present in various dietary supplements and generally promoted as an accepted alternative to estrogen replacement therapy. Potential isoflavone biomedical exploitation is not only limited to estrogen replacement therapy, so it should be treated in a wider context of different ageing symptoms remediation.

Primary Neuroendocrine Tumor in the Liver

Neuroendocrine tumors (NETs) are rare and typically have an indolent course. NETs usually arise from pancreas or small bowel and metastasize to the liver. Only 25% of NETs are hormone producing. Primary hepatic NETs are very rare, accounting for 0.3% of all NETs. As such, the clinical presentation is not well recognized. The following case describes the presentation of a primary hepatic NET and management of its complications. A 44 year-old male presented with 1 week of right upper quadrant pain and jaundice. He reported a month long history of dyspepsia refractory to PPI, early satiety, and 10-pound weight loss. He had evidence of hepatocellular and cholestatic liver injury (Table 1). Imaging showed biliary obstruction due to a 5 cm x 10 cm x 4 cm heterogeneous, multicystic, enhancing mass in the left hepatic lobe. Ultrasound suggested hepatocellular carcinoma, while CT and MRI raised concern for intraductal lesions and cholangiocarcinoma (Figures 1-2). On day 2, ERCP identified 2 filling defects in the common bile duct (CBD), likely causing intermittent obstruction. CBD brushings were negative for malignant cells. The obstruction was treated with biliary sphincterotomy and placement of a 8.5 Fr by 9 cm stent into the CBD. On day 4, EUS showed a mixed cystic and solid mass lesion in the left hepatic lobe along with a 4 cm long intraductal mass - of the same density - extending from the common hepatic duct to the CBD. The left intrahepatic bile ducts were diffusely dilated around the hepatic mass. The common hepatic duct and CBD were dilated to 18mm secondary to the intraductal mass. On day 5, ERCP was repeated due to evidence of persistent biliary obstruction on EUS and fluctuating bilirubin (Table 1). This second ERCP showed filling defects at the hepatic duct bifurcation and proximal CBD. The left hepatic duct was completely obstructed (Figure 3). The previous biliary stent was replaced with an 8.5 Fr by 15 cm stent into the right hepatic duct, and biliary obstruction was relieved. Biopsies from the EUS showed a low-grade NET. Chromogranin A was elevated at 2111 ng/mL and 5-HIAA was within the reference range. The patient was discharged with plans for an octreotide scan and follow up with surgical oncology.Figure: Hepatic Mass on CT.Figure: Hepatic Mass on T2 weighted MRI.Figure: Biliary Tree on ERCP #2, with complete obstruction of left hepatic duct.Table: Table. Liver Panel Through Treatment CourseNETs are rare and can have a variety of presentations, based on their origin and activity. This case highlights one of the few documented occurrences of biliary obstruction as the presenting feature of a primary hepatic NET.

Discovering potential Streptomyces hormone producers by using disruptants of essential biosynthetic genes as indicator strains.

Autoregulators are low-molecular-weight signaling compounds that control the production of many secondary metabolites in actinomycetes and have been referred to as 'Streptomyces hormones'. Here, potential producers of Streptomyces hormones were investigated in 40 Streptomyces and 11 endophytic actinomycetes. Production of γ-butyrolactone-type (IM-2, VB) and butenolide-type (avenolide) Streptomyces hormones was screened using Streptomyces lavendulae FRI-5 (ΔfarX), Streptomyces virginiae (ΔbarX) and Streptomyces avermitilis (Δaco), respectively. In these strains, essential biosynthetic genes for Streptomyces hormones were disrupted, enabling them to respond solely to the externally added hormones. The results showed that 20% of each of the investigated strains produced IM-2 and VB, confirming that γ-butyrolactone-type Streptomyces hormones are the most common in actinomycetes. Unlike the γ-butyrolactone type, butenolide-type Streptomyces hormones have been discovered in recent years, but their distribution has been unclear. Our finding that 24% of actinomycetes (12 of 51 strains) showed avenolide activity revealed for the first time that the butenolide-type Streptomyces hormone is also common in actinomycetes.

Regulation of pituitary stem cells by epithelial to mesenchymal transition events and signaling pathways

The anterior pituitary gland is comprised of specialized cell-types that produce and secrete polypeptide hormones in response to hypothalamic input and feedback from target organs. These specialized cells arise from stem cells that express SOX2 and the pituitary transcription factor PROP1, which is necessary to establish the stem cell pool and promote an epithelial to mesenchymal-like transition, releasing progenitors from the niche. The adult anterior pituitary responds to physiological challenge by mobilizing the SOX2-expressing progenitor pool and producing additional hormone-producing cells. Knowledge of the role of signaling pathways and extracellular matrix components in these processes may lead to improvements in the efficiency of differentiation of embryonic stem cells or induced pluripotent stem cells into hormone producing cells in vitro. Advances in our basic understanding of pituitary stem cell regulation and differentiation may lead to improved diagnosis and treatment for patients with hypopituitarism.

Evaluation of endophytic bacteria from some rice varieties for plant growth promoting activities

Eighteen endophytic bacteria were isolated from different rice varieties by using Jensen’s nitrogen free medium. According to morphological and biochemical tests, isolates S1, R1, R2, R5 and R6 may be Rhizobium spp. and the remaining isolates may be Azospirillum spp. Fourteen out of isolates had plant growth hormone (Indole Acetic Acid) producing activity. R7 was the best Indole Acetic Acid producer among the isolated strains at two days incubation period in Jensen’s nitrogen free medium with L-tryptophan 0.5mg /ml at 37˚C and the concentration produced was 120.55 ppm. In nitrogen free semi-solid agar medium containing trace amount of bromothymol blue as indicator, all isolates showed nitrogen fixing activity during seven days incubation period. All isolates except S6 could solubilize phosphate on Pikovskaia’s medium supplemented with (0.5% Ca3(PO4)2). S5 was the best P- solubilizer at 5 days incubation period both in plate screening showing zone diameter 45mm and 539.78 ppm quantitatively. Fourteen isolates except (LS, R2, S4, S8) could also decompose potassium (ranging from 2-30mm in diameter) on K- decomposing medium with 0.12% Kmica. R3 was the best K-decomposer by plate screening (30mm in diameter) and 19.84 ppm quantitatively. S3 and R3 had good antagonistic activities against Rhizotonia solani and LS and R5 to Phythium spp. and S7 to Fusarium oxysporum respectively.

Altered lipid metabolism in Drosophila model of Huntington's disease.

Huntington’s disease (HD) is late-onset, progressive neurodegenerative disorder caused by expansion of polyglutamine (polyQ) repeat within Huntingtin (Htt) protein. In HD patients, energy-related manifestations such as modulation of weight during entire course of disease with energy deficit at terminal stage have been reported, however, underlying reason remains elusive till date. Lipids, carbohydrate and protein constitute a predominant fraction of body’s energy reservoir and perturbation in their homeostasis may influence weight. To discern role of these energy molecules in weight alteration, we quantified them in an in vivo transgenic Drosophila model of HD. We document that diseased flies exhibit change in weight due to an altered lipid metabolism, as evident from considerably high lipid levels at the time of disease onset followed by a pathologic decline at end-stage. An alteration in intracellular lipid droplet size suggested altered cellular lipid turnover. Furthermore, diseased flies displayed substantial changes in carbohydrate and protein content. Interestingly, alteration in weight and lipid levels are independent of the feeding pattern in diseased condition and exhibit weak correlation with insulin-like peptide or adipokinetic hormone producing cells. We propose that therapeutic intervention aimed at restoring lipid levels and associated metabolic pathways may improve longevity and quality of patient’s life.

Microscopic venous invasion in patients with pancreatic neuroendocrine tumor as a potential predictor of postoperative recurrence

In-vivo cell kinetics study following peroperative intravenous infusion of bromodeoxyuridine (BrdU) was done in 10 cases of primitive neuroectodermal tumors (PNET) and 44 nonglial tumors of different histological types. The histological features usually regarded as indicators of aggressive behaviour were examined in these tumors and correlated with in-vivo labelling index (LI). In the case of meningiornas, increased cellularity, pleomorphism and mitosis had no correlation with LI. Benign non-recurrent meningiomas usually showed LI 5 1% (mean 0.6 ± 0.3%) whereas recurrent and malignant meningioma showed higher LI (mean 2.6 ± 0.5% and 2.8 ± 0.4% respectively). Follow-up study suggested that meningiomas having benign histological appearance with LI > 1% might have increased chance of recurrence. In cases of PNETs among different histological features, mitosis and differentiation seemed to be related to the biological behaviour. Astrocytic differentiation was associated with lower rate of proliferation. In pituitary adenomas different hormone producing and null cell adenomas showed similar low proliferative potential (0.6 ± 0.3%). Benign nerve sheath tumors, craniopharyngioma and choroid plexus papilloma showed low in-vivo LI of less than 1%.Thus the present study revealed the inadequacies of routine histological examination in assessing the aggressiveness of the nonglial tumors, especially rneningiomas. In-vivo LI may be a good supplement to histological diagnosis as well as helping to assess the prognosis and accordingly the management of individual cases.

Isolation of Phosphate Solubilizing Bacteria from Maize Rhizosphere and Their Potential for Rock Phosphate Solubilization–Mineralization and Plant Growth Promotion

ABSTRACTPhosphate solubilizing bacteria (PSB) play a significant role in plant P nutrition by their effect on soil P dynamics and their subsequent ability to make P available to plants via solubilization and mineralization processes. This study aimed to evaluate the effect of separate and combined use of indigenous PSB, poultry manure (PM) and compost on solubilization and mineralization of rock phosphate (RP) and their subsequent effect on growth and P accumulation of maize (Zea mays L.). A group of fifty seven bacteria were isolated from the rhizosphere/rhizoplane of maize that had been grown in soils collected from varying altitudes (655–2,576 m) of the mountain region of Rawalakot, Azad Jammu and Kashmir, Pakistan. After screening, the capacity of eleven isolates to solubilize mineral phosphate was quantitatively evaluated using insoluble Ca3(PO4)2 in culture medium as a time course study through spectrometer. The growth hormone producing (IAA) capacity of the isolates was also determined. Furthermore, five potential isolates were tested for their ability to increase P release capacity (mineralization) of insoluble RP in an incubation study. The effect of PSB inoculation on maize was determined in a completely randomized greenhouse experiment where root and shoot biomass and P accumulation in plants were assessed. The P solubilization index of selected isolates varied from 1.94 to 3.69, while the P solubilization efficiency ranged between 94.1% and 269.0%. The isolates MRS18 and MRS27 displayed the highest values. The P solubilization in the liquid medium was maximum at 6 and 9 days of incubation ranging between 9.91 and 44.04 µgmL−1 and the isolates MRS27 and MRS34 exhibited the highest solubilization. Six isolates showed additional capability of producing IAA ranging between 2.66 and 28.41 µgmL−1. Results of the incubation study indicated that P release capacity (P mineralization) of RP-amended soil varied between 6.0 and 11.8 µgPg−1 that had been significantly increased to 30.6–36.3 µgPg−1 (maximum value) when PSB were combined with RP. The combined application of PSB and organic amendments (PM, compost) with RP further increased P mineralization by releasing a maximum of 37.7 µgPg−1 compared with separate application of RP (11.8 µgPg−1) and organic amendments (21.5 and 16.5 µgPg−1). The overall effect of PSB (as a group) with RP over RP alone on maize growth showing a relative increase in shoot length 21%, shoot fresh weight 42%, shoot dry weight 24%, root length 11%, root fresh weight 59%, root dry weight 35% and chlorophyll content 32%. This study clearly indicates that use of PSB, and organic amendments with insoluble RP could be a promising management strategy to enhance P availability in soil pool and improve plant growth in intensive cropping systems.

Hedgehog signaling activation induces stem cell proliferation and hormone release in the adult pituitary gland.

Hedgehog (HH) signaling is known to be essential during the embryonal development of the pituitary gland but the knowledge about its role in the adult pituitary and in associated tumors is sparse. In this report we investigated the effect of excess Hh signaling activation in murine pituitary explants and analyzed the HH signaling status of human adenopituitary lobes and a large cohort of pituitary adenomas. Our data show that excess Hh signaling led to increased proliferation of Sox2+ and Sox9+ adult pituitary stem cells and to elevated expression levels of adrenocorticotropic hormone (Acth), growth hormone (Gh) and prolactin (Prl) in the adult gland. Inhibition of the pathway by cyclopamine reversed these effects indicating that active Hh signaling positively regulates proliferative processes of adult pituitary stem cells and hormone production in the anterior pituitary. Since hormone producing cells of the adenohypophysis as well as ACTH-, GH- and PRL-immunopositive adenomas express SHH and its target GLI1, we furthermore propose that excess HH signaling is involved in the development/maintenance of hormone-producing pituitary adenomas. These findings advance the understanding of physiological hormone regulation and may open new treatment options for pituitary tumors.

Diet-Induced Obesity and Ghrelin Effects on Pituitary Gonadotrophs: Immunohistomorphometric Study in Male Rats.

The close relationship between energy metabolism, nutritional state, and reproductive physiology suggests that nutritional and metabolic disorders can disrupt normal reproductive function and fertility. Considering the importance of leptin and ghrelin effects in regulation of the hypothalamic-pituitary-gonadal axis, the objective of this study was to investigate the influence of obesity and centrally applied ghrelin on immunohistochemical appearance and quantitative morphology of the pituitary follicle-stimulating hormone (FSH) and luteinizing hormone (LH) producing cells in adult male rats. In this experimental study, animals were given two differ- ent diets: normal-fat (NF) and high-fat (HF), for 4 weeks, corresponding to normal and positive energy balance (n=2×14), respectively. Each group was subsequently divided into two subgroups (n=7) receiving intracerebroventricular (ICV) injections of either ghrelin [G, 1 µg/5 µL phosphate buffered saline (PBS)] or vehicle (5 µL PBS, control group) every 24 hours for five consecutive days. Morphometric analyses showed that in HF control group, the percentage of FSH cells per unit volume of total pituitary gland tissue (in μm(3)), i.e. volume density (Vvc), was increased (P<0.05) by 9.1% in comparison with the NF controls. After ICV treatment with ghrelin, volume (Vc) and volume density (Vvc) of FSH cells in ghrelin+NF (GNF) and ghrelin+HF (GHF) groups remained unchanged in comparison with NF and HF controls. Volume of LH cells in HF control group was increased by 17% (P<0.05), but their Vvc was decreased by 8.3% (P<0.05) in comparison with NF controls. In GNF group, the volume of LH cells increased by 7% (P<0.05), in comparison with the NF controls, but in GHF group, the same parameter remained unchanged when compared with HF controls. The central application of ghrelin de- creased the Vvc of LH cells only in GNF group by 38.9% (P<0.05) in comparison with the NF control animals. The present study has shown that obesity and repetitive ICV administra- tion of low doses of ghrelin, in NF and HF rats, modulated the immunohistomorphometric features of gonadotrophs, indicating the importance of obesity and ghrelin in regulation of the reproductive function.

Implications of Hydrogen Sulfide in Glucose Regulation: How H2S Can Alter Glucose Homeostasis through Metabolic Hormones.

Diabetes and its comorbidities continue to be a major health problem worldwide. Understanding the precise mechanisms that control glucose homeostasis and their dysregulation during diabetes are a major research focus. Hydrogen sulfide (H2S) has emerged as an important regulator of glucose homeostasis. This is achieved through its production and action in several metabolic and hormone producing organs including the pancreas, liver, and adipose. Of importance, H2S production and signaling in these tissues are altered during both type 1 and type 2 diabetes mellitus. This review first examines how H2S is produced both endogenously and by gastrointestinal microbes, with a particular focus on the altered production that occurs during obesity and diabetes. Next, the action of H2S on the metabolic organs with key roles in glucose homeostasis, with a particular focus on insulin, is described. Recent work has also suggested that the effects of H2S on glucose homeostasis goes beyond its role in insulin secretion. Several studies have demonstrated important roles for H2S in hepatic glucose output and adipose glucose uptake. The mechanism of H2S action on these metabolic organs is described. In the final part of this review, future directions examining the roles of H2S in other metabolic and glucoregulatory hormone secreting tissues are proposed.