Genetic Structure and Nucleotide Sequences of Three Productivity Trait Genes, in Egyptian Buffalo

One of the main sources of meat and milk in Egypt is buffalo, of river type. The marker assisted selection-depending on the promised genetic markers is considered to be the effective way for improvement of buffalo productivity. This work aimed to study the genetic structure and nucleotide sequences of three productivity genes namely; LGB, PIT-1, and POUF-1 in Egyptian buffalo. This study is performed by using genomic DNA, which was extracted from 100 female buffaloes. The DNA extracts were subjected to PCR by using some specific primers of the tested genes. The PCR products were digested with dedicated restriction enzymes like; HaeIII for LGB and HinfI for both PIT-1 and POUF-1 genes. Depending on the appearance of restriction sites in the amplified fragments; GG

Acromegaly due to a Macroinvasive Plurihormonal Pituitary Adenoma and a Rectal Carcinoid Tumor

A macroinvasive pituitary adenoma with plurihormonality usually causes acromegaly and hyperprolactinemia, and also accompanies with neurologic symptoms such as visual disturbances. However, its concurrent presentation with a rectal carcinoid tumor is rarely observed. This study reports the history, biochemical, colonoscopic and immunohistochemical results of a 48-year-old female with acromegaly and hyperprolactinemia. Despite the large size and invasive nature of the pituitary adenoma to adjacent anatomical structures, she did not complain of any neurologic symptoms such as visual disturbance or headache. Immunohistochemical staining of the surgical specimen from the pituitary adenoma revealed that the tumor cells were positive for growth hormone (GH), prolactin (PRL), and thyroid stimulating hormone (TSH). Staining for pituitary-specific transcription factor-1 (Pit-1) was shown to be strongly positive, which could have been possibly contributing to the plurihormonality of this adenoma. Colonoscopy found a rectal polyp that was identified to be a carcinoid tumor using immunohistochemical staining. A macroinvasive pituitary adenoma with concomitant rectal carcinoid tumor was secreting GH, PRL, and TSH, which were believed to be in association with over-expression of Pit-1. This is the first case report of double primary tumors comprising a plurihormonal pituitary macroadenoma and rectal carcinoid tumor.

Efeitos da sobrecarga de fósforo dietético na expressão dos cotransportadores NaP-IIb e PiT-1 em ratos controles e urêmicos / Phosphorus overload effects on NaP-IIb and PiT-1 cotransporters in control and uremic rats

O fósforo é um dos minerais mais abundantes no corpo além de ser essencial para muitos processos biológicos. A homeostase do fósforo depende da absorção no intestino, da excreção renal, da remodelação óssea e de hormônios como o paratormônio, calcitriol e FGF-23. Nos pacientes com doença renal crônica, a excreção urinária de fósforo está comprometida levando à hiperfosfatemia, que contribui para o aumento da morbidade e mortalidade desses pacientes. A absorção intestinal de fósforo no intestino delgado, particularmente via cotransportadores NaP-IIb e PiT-1, é pouco estudada. O objetivo desse estudo foi avaliar os efeitos de dietas com diferentes concentrações de fósforo na expressão proteica e gênica dos cotransportadores NaP-IIb e PiT-1, bem como na apoptose dos enterócitos dos diferentes segmentos do intestino delgado de animais controles e urêmicos. Estudamos setenta e seis ratos Wistar machos, inicialmente divididos em dois grupos: controles (C) e urêmicos (Nx). Cada grupo foi subdividido em outros três, de acordo com a concentração de fósforo (P) na dieta: dieta Baixa (0,2%), dieta Padrão (0,54%) e dieta Alta (0,9%). Analisamos parâmetros bioquímicos (creatinina, P, Cai, PTH e FGF-23), a expressão proteica dos cotransportadores, através de Western Blotting, ELISA e imunofluorescência, a expressão gênica por PCR em tempo real e a apoptose dos enterócitos pela técnica TUNEL. Os resultados mostraram que os níveis séricos de creatinina, P, PTH e FGF-23 foram significativamente mais elevados nos animais Nx. Nos animais C com dieta baixa em P observamos aumento da expressão proteica do cotransportador NaP-IIb em todos os segmentos do intestino enquanto, no Nx Baixo, a expressão desse cotransportador foi menor somente no jejuno. Quanto ao PiT-1, sua expressão foi menor no íleo do grupo Nx Alto comparado ao seu respectivo controle. A expressão gênica do cotransportador NaP-IIb foi menor no jejuno do Nx Alto e maior no íleo desse mesmo grupo em relação aos seus...

Phosphorus is one of the most abundant minerals in the body besides being essential for many biological processes. Phosphorus homeostasis depends on absorption in the small intestine, renal excretion, bone remodeling and hormones such as parathyroid hormone, calcitriol and FGF-23. In patients with chronic kidney disease phosphorus urinary excretion is compromised leading to hyperphosphatemia, which contributes to increased morbidity and mortality of these patients. Intestinal absorption of phosphorus in the small intestine, particularly of NaP-IIb and PiT-1 cotransporters is poorly studied. The aim of this study was to evaluate the effects of diets with different concentrations of phosphorus in protein expression and gene of NaP-IIb and PiT-1 cotransporters as well as in apoptosis of enterocytes of different segments of intestine in control and uremic animals. We studied seventy-six male Wistar rats initially divided into two groups: controls (C) and uremic (Nx). Each group was subdivided into three others, according to the phosphorus concentration in the diet: Low diet (0.2% P), Standard diet (0.54% P) and High diet (0.9% P). We analyzed biochemical parameters (creatinine, P, iCa, PTH and FGF-23), protein expression of cotransporters, using Western Blot, ELISA, immunofluorescence, real-time PCR and apoptosis of enterocytes using the TUNEL technique. Results showed that serum creatinine, P, PTH and FGF-23 were significantly higher in Nx animals. In C animals with a diet low in P we observed increase in protein expression of NaP-IIb cotransporter in all segments of the intestine while in low Nx expression of this cotransporter was lower only in jejunum. As for PiT-1 expression was lower in the ileum of the high Nx group compared to their respective control. Gene expression of NaP-IIb cotransporter was lower in the jejunum of high Nx and higher in the ileum of the same group as compared to their respective controls. PiT-1 gene expression was higher in all...

Effect of bromocriptine on the expression of Pit-1 in prolactinoma rats / 中国药学杂志

OBJECTIVE: To investigate the effect of bromocriptine on the expression of Pit-1 in prolactinoma rats. METHODS: Firstly, to prepare prolactinoma model in rats. Adult Wistar rats were divided into two groups at random. The rats in control group were subscutaneously implanted with a blank implant. Rats in 17β-estradiol group were implanted with 17β-estradiol-containing implants. Secondly, rates in 17β-estradiol group were divided into two groups at random: model group and bromocriptine group. Water was administrated to rats in model group. Bromocriptine (0.225 mg · kg-1 · d-1) were orally administrated to rats in bromocriptine group, the rats in control group were orally administrated with water. After four weeks of treatment, all the animals were executed. Each pituitary gland was weighed. Serum prolactin (PRL) levels were measured by RIA method. Pit-1 mRNA levels in pituitaiy tissue were measured by RT-PCR method. RESULTS: The weights of pituitary gland and PRL levels in 17β-estradiol group were individually higher than those in control group (P<0.001). The expression levels of Pit-1 mRNA in bromocriptine group was obviously lower than that in model group (P≤0.001). CONCLUSION: Bromocriptine has potential preventive effect to estrogen-induced rat prolactinoma. The decrease of Pit-1 mRNA level may be involved in the mechanism of anti-prolactinoma effect of bromocriptine.

Cytokines regulate the promoter activities of human growth hormone gene in rat pituitary MtT/S cells / 中国免疫学杂志

Objective:To study the effect of interleukin-11(IL-11),ciliary neurotropic factor (CNTF) and transforming growth factor-β (TGF-β) on the hGH gene promoter activity in rat pituitary MtT/S cells and the interaction with pituitary-specific transcription factor Pit-1.Methods:Stable transformed MtT/S cell line which contains hGH gene promoter -484-30 bp and luciferase reporter gene firstly established,then the concentration of GH in the medium and lysate of MtT/S cells and luciferase activities in MtT/S cells were measured after treatment these cells with the above cytokines,the effects of cytokines on secretion and synthesis of GH,and the promoter activity of the hGH gene were observed.Results:The results showed that IL-11(20 nmol/L),CNTF(10 nmol/L) and TGF-β(5 nmol/L) regulated secretion and synthesis of GH,and the luciferase expression in stable transformed MtT/S cells.IL-11 and CNTF had the stimulatory effect,whereas TGF-β had the inhibitory effect.Neither overexpression of Pit-1 nor inhibiting Pit-1 expression affected the regulatory role of these cytokines.Conclusion:IL-11,CNTF and TGF-β regulate the GH production in pituitary MtT/S cell line by regulating the hGH gene promoter activity.Pit-1 may not be involved in these actions.

Análise da expressão dos genes PROP1 e CTNNB1 em craniofaringiomas adamantinomatosos com e sem mutação somática no CTNNB1 / Analysis of PROP1 and CTNNB1 expression genes in adamantinomatous craniopharyngiomas with and without CTNNB1 somatic mutation

Os craniofaringiomas são os tumores mais frequentes da região hipotálamohipofisária na faixa etária pediátrica. Apesar de serem histologicamente benignos, sua tendência infiltrativa e seu comportamento agressivo resultam em significante morbimortalidade. Histologicamente podem ser divididos em dois subtipos: adamantinomatosos e papilíferos. A patogênese dos craniofaringiomas é pouco compreendida. Mutações no gene CTNNB1, que codifica a proteína beta-catenina, são a única alteração molecular conhecida até o momento implicada na tumorigênese dos craniofaringiomas adamantinomatosos. Tais mutações afetam o sítio de degradação da beta-catenina, que passa a se acumular no citoplasma e no núcleo, ativando excessivamente a via de sinalização WNT, através da ligação aos fatores de transcrição da família LEF/TCF, levando a tumorigênese. Recentemente foi descoberto um novo mecanismo de determinação da linhagem celular hipofisária regulado pela beta-catenina, através do qual ela interage diretamente com o PROP1 para determinar a diferenciação celular hipofisária. De acordo com esse modelo, o complexo protéico PROP1/beta- catenina atua simultaneamente como repressor do HESX1 e ativador do PIT1, dependendo dos co-fatores associados. Pacientes com mutações germinativas inativadoras no PROP1 desenvolvem hipopituitarismo e podem apresentar aumento hipofisário com imagens de ressonância nuclear magnética (RNM) da região selar muitas vezes semelhantes àquelas dos craniofaringiomas, com hiperssinal em T1. Por outro lado, camundongos com expressão persistente do Prop1 exibem defeitos na regulação da proliferação celular hipofisária, incluindo cistos da bolsa de Rathke, hiperplasia adenomatosa e tumores, sugerindo que mutações com ganho de função no PROP1 também poderiam contribuir para a patogênese de tumores hipofisários em seres humanos. A semelhança entre as imagens de RNM dos pacientes com craniofaringiomas e daqueles com aumento hipofisário devido a mutações...

Craniopharyngiomas are the the commonest tumors to involve the hypothalamo-pituitary regions in childhood population. Histologically they are benign, and can be divided in two primary subtypes: the adamantinomatous and the papillary. Although histologically benign, their infiltrative tendency and aggressive behavior can result in great morbidity. The pathogenesis of craniopharyngiomas is poorly understood. To date, beta-catenin gene (CTNNB1) mutations have been identified only in the adamantinomatous subtype. These mutations affect the degradation target box of beta-catenin that accumulates in the cytoplasm and the nucleus increasing the transcriptional activity of WNT pathway through interaction with the transcription factors of LEF/TCF family, leading to tumorigenesis. Recently, an interaction between beta-catenin and PROP1 was described as a new mecanism for beta-catenindependent regulation of pituitary cell-lineage determination. According to this novel model, the PROP1/beta-catenin proteic complex would act as a binary switch to simultaneously repress the transcription factor HESX1 and to activate expression of transcription factor PIT1, depending on the associated cofactors. Patients with loss-of-function mutations in PROP1 present combined pituitary hormonal deficiency generally associated with pituitary enlargement and the magnetic resonance imaging (MRI) of the sellar region in these patients sometimes resembles that of the craniopharyngiomas, with T1 hyperintense signal. On the other hand, transgenic mice with persistent Prop1 expression exhibit defects consistent with misregulation of pituitary cell proliferation, including adenomatous hyperplasia with formation of Rathke's cleft cysts and tumors suggesting that misregulation of PROP1 expression in human could contribute to pathogenesis of pituitary tumors. The similarity between the MRI images of craniopharyngiomas patients and that of patients with loss-of-function mutations in...

Effects of Cadmium on Placental Function and Reproduction in Rats / 대한산업의학회지

OBJECTIVES: This study investigated the toxic effects of cadmium on placental function and reproduction in rats. For this study, the mRNA levels of the placental prolactin-growth hormone (PRL-GH) gene family, placental trophoblast cell frequemcy and reproductive data were analyzed. METHODS: Pregnant F344 Fisher rats (200 g+/-23 g) were intraperitoneally injected with 0, 0.5, and 5.0 mg/kg B.W/day of cadmium (CdCl2) dissolved in saline from days 7-11 or 16-20 of pregnancy, and were sacrificed at days 11 or 20, respectively. The mRNA levels were analyzed by Northern blot hybridization and reverse transcription-polymerase chain reaction. The hormone concentration was analyzed by radioimmunoassay and the frequemcy of the placental trophoblast cells was observed by histochemical study. Reproductive data were surveyed at day 20 of the pregnancy and after the births. Statistical analysis was carried out using the SAS program (version 8.1). RESULTS: The mRNA levels of the PRL-GH gene family were reduced dose dependently by cadmium. The mRNA levels of Pit-1a and -b isotype genes were also reduced by cadmium. The hormone concentration of PL-Iv and -II was decreased by cadmium. During the second half of pregnancy (days 11-21), a high dose of cadmium exposure significantly reduced the frequency of spongiotrophoblast and trophoblast giant cells that secrete the PRL-GH hormones. In the last stage of pregnancy (day 20), a high dose of cadmium exposure induced the apoptosis of spon-giotrophoblast cells in the junctional zone of the placenta. Reproductive data such as placental and infant weight, number of live fetuses were decreased, and number of resorptions and dead fetuses, post-implantation loss were increased significantly in the cadmium exposed group compared with the control. CONCLUSIONS: Cadmium disrupts the functions of the placenta and these effects leads to reproductive disorders in rats.

Effects of Toluene on the Expression of Placental PRL-GH Family Genes and Reproduction in the Rat / 대한산업의학회지

OBJECTIVES: The purpose of this experimental study was to investigate the toxic effects of toluene on the placental functions and reproductionin the rat. In this study, the expression of placental prolactin-growth hormone (PRL-GH) and Pit-1 genes, the frequency of placental trophoblast cells, and the reproductive data were analyzed. METHODS: The pregnancy of the Sprague-Dawley rats (250+/-25 g) was determined by verifying the presence of the copulatory plug or sperm in the vaginal smear and the day on which this was observed was defined as pregnancy day 0. The pregnant rats were divided into three groups. The control group was intraperitoneally (ip) injected with sesame oil, and the other two groups were given either 150 or 750 mg/kg BW/day of toluene resuspended in sesame oil during pregnancy days 7-11 and 16-20. The rats from the three experimental groups were sacrificed on pregnancy days 11 and 20, respectively. The mRNA levels of the PRL-GH, Pit-1a and b isotype genes were analyzed by Northern blot hybridization and Reverse transcription-polymerase chain reaction (RT-PCR), respectively. The hormonal concentration was analyzed by Radioimmunoassay. The frequency of the placental trophoblast cells was determined by means of a histochemical study. Reproductive data, such as the placenta and infnat weight, pregnancy period and litter size were surveyed at pregnancy day 20 and after birth. Statistical analysis was carried out by means of the SAS program (version 8.1). RESULTS: The mRNA levels of the PRL-GH family genes were reduced in a linear fashion by exposure to toluene. The mRNA levels of the Pit-1a and b isotype genes, which induce the expression of the PRL-GH family genes, were also reduced by exposure to toluene. The placental lactogen Iv and II concentrations in the rat placenta, fetus and maternal blood were also decreased by exposure to toluene. During the last stage of gestation, exposure to a high dose of toluene reduced the frequency of the spongiotrophoblast cells that secrete the PRL-GH hormones. Reproductive data such as the placenta and infant weight, and litter size were reduced, and the pregnancy period was extended in the toluene exposed group as compared with the control group. CONCLUSIONS: Toluene disrupts the PRL-GH hormone metabolism in the rat placenta and this leads to reproductive disorder.

Gene Expression of Somatostatin Receptor (Subtype 2 & 5), Gi2 alpha and Pit-1 in GH-secreting Pituitary Adenomas / 대한내분비학회지

BACKGROUND: Mutation of Gs protein subunit (gsp oncogene), detected in about 30~40% of growth hormone (GH)-secreting pituitary tumors, is associated with an increased long-acting somatostatin analog octreotide sensitivity. However, the mRNA expression of somatostatin receptor (sst) was not changed in the GH-secreting pituitary tumor, regardless of whether they were gsp oncogene positive or negative. This suggests that the expression of genes coding for Gi2 alpha , Pit-1 and the other factors involved in the regulation of secretory activity in somatotrophs is likely to be altered in gsp oncogene positive tumors. We observed the impact of the gsp oncogene on the expression of the genes coding for Gi2 alpha, Pit-1 and sst (2&5) in GH-secreting pituitary tumors. METHODS: The GH response to octreotide was examined in 13 acromegalic patients before transsphenoidal adenomectomy. Genomic DNA and RNA were extracted from fresh frozen tumor tissues. PCR was performed to amplify and sequence the region between codon 184 and 251 that includes exons 8 and 9 of the Gs gene. Sst2, sst5, Gi2 alpha and Pit-1 mRNA levels were measured by semi-quantitative RT-PCR. RESULTS: Sst2 and sst5 mRNA transcripts were detected in all tumors (7 gsp +, 6 gsp-). The amount of sst transcripts varied considerably varied between the tumors. There were no significant differences in sex, age, tumor size, grade or basal GH levels. Pit-1 and sst2 mRNA levels were not different. In contrast, Gi2 alpha mRNA levels were significantly higher in gsp (+) while sst5 mRNA levels were higher in gsp (-). CONCLUSION: These data suggests that gsp oncogene may increase Gi2 alpha levels but decrease sst5 mRNA levels. However, Pit-1 and sst2 mRNA expression may not be affected by gsp oncogene. The increased expression of the Gi2 alpha gene might be an inhibitory compensatory response to the action of gsp oncogene.

Gene Expression of the Somatostatin Receptors, Gi2 alpha and Pit-1 alpha in GH3 Cells Permanently Transfected with a Mutant Gs alpha Gene / 대한내분비학회지

BACKGROUND: Cyclic AMP stimulates the expression of the somatostatin (SRIF) receptor (sst1-5) and human growth hormone (GH)-secreting pituitary tumors with the gsp oncogene which increases intracellular cAMP levels, and shows a good inhibitory response of the GH to SRIF. Taken together, we hypothesized that the gsp oncogene may increase the SRIF receptor expression or and factors related to the postreceptor signal transduction of the SRIF, in order to enhance its responsiveness to SRIF. To test this hypothesis, we investigated if the gsp oncogene could increase the sst1, sst2, Gi2 alpha, and pit-1 alpha gene expression in GH3 cells. METHODS: GH3 cells were permanently transfected with the plasmid expressing Gs alpha gene, where the arginine of codon 201 was replaced with histidine. Intracellular cAMP levels and GH concentrations were measured by radioimmunoassays. Gene expressions of the sst1, sst2, Gi2 alpha, and pit-1 alpha were determined by RT-PCR. RESULTS: Intracellular cAMP levels and medium GH release were increased by 1.7 and 2.7-fold in GH3 cells expressing the gsp oncogene, respectively. In GH3 cells expressing the gsp oncogene, the sst1 mRNA levels were decreased, whereas those of the sst2, Gi2 alpha and pit-1 alpha mRNA were increased. A 4-h forskolin (10 M) stimulation remarkably increased the sst1 and sst2 mRNA levels in GH3 cells expressing wild and mutant Gs alpha . However, forskolin did not affect the Gi2 alpha and pit-1 alpha mRNA levels. In contrast, SRIF (1 M, 2 h) decreased the sst2 mRNA levels only in GH3 cells expressing the gsp oncogene. CONCLUSION: These results suggest that higher expressions of sst2, Gi2 alpha, and pit-1 alpha, induced by the gsp oncogene may be a mechanism by which gsp-positive pituitary tumors show a greater response to SRIF. The discrepancy between these and in vivo results should be explored further.

Effects of Toluene, Xylene and Trichloroethylene on the Regulation of GnRH, GnRH Receptor and Pit-1 Gene Expression in Male Rat Hypothalamus and Pituitary / 대한산업의학회지

The workplace exposure of chemicals has steadily increased, therefore the concern for subsequent effect on reproductive outcome has been an important issue in occupational medicine. In previous studies, higher rates of spontaneous abortion, reduced fertility and menstrual disorder among women, and an impairment of sperm quantity and quality among men have been associated with a wide variety of chemical agents. This study was conducted to evaluate the effects of toluene, xylene and trichloroethylene (TCE) injection on the mRNA levels of GnRH, GnRH receptor and Pit-1 genes in male rats hypothalamus and pituitary and the effects on the plasma levels of FSH, LH, prolactin and testosterone. Sprague-Dawley male rats were divided into five groups of five each according to concentration of toluene, xylene and TCE. The rats were injected subcutaneously to 0, 50, 100, 200, 400 mg/kg body weight/day of toluene, xylene and TCE, respectively for 6 days. Rat brains were excised and hypothalamus and pituitary were separated. Reverse transcription-polymerase chain reaction (RT-PCR) and RNase protection assay (RPA) were used to evaluate the GnRH, GnRH receptor and Pit-1 mRNA levels. Plasma concentrations of FSH, LH, prolactin and testosterone were assayed by radioimulunoassay (RIA). The results were as follows; 1. GnRH, GnRH receptor and Pit-1 mRNA levels in toluene and xylene injected groups, and GnRH receptor mRNA levels in TCE injected group were lowered dose-dependently. Especially, GnRH receptor and Pit-1 mRNA levels in 200 mg/kg of toluene injected group, and GnRH, GnRH receptor and Pit-1 mRNA levels in 400 mg/kg of toluene injected group were significantly lowed than control group (p<0.05). GnRH receptor and Pit-1 mRNA levels in 400 mg/kg of xylene injected group, and GnRH receptor mRNA levels in 400 mg/kg of TCE injected group were significantly lower than control group (p<0.05). 2. The plasma levels of prolactin and testosterone in 400 mg/kg of toluene injected group, and LH in 100, 200 and 400 mg/kg of xylene injected group, and testosterone in 400 mg/kg of TCE injected group were significantly lower than control group (p<0.05). In conclusion, we speculated that toluene and xylene affected reproductive system secondarily through hypothalamus-pituitary axis, and TCE affected directly through steroidogenesis. And we recomended that further study for assessment of the reproductive toxiclty of mixed organic solvent exposures should be conducted.