In vivo and in vitro regulation of Akt activation in human endometrial cells is estrogen dependent

Estrogen-bound estrogen receptors (ER) alpha and beta classically activate gene expression after binding to the estrogen response element in the promoter regions of target genes. Estrogen also has rapid, nongenomic effects. It activates several membranous or cytoplasmic kinase cascades, including the phosphatidylinositol 3-phosphate (PI3K/Akt) cascade, a signaling pathway that plays a key role in cell survival and apoptosis. Normal human endometrium is exposed to variable levels of steroid hormones throughout the menstrual cycle. We hypothesized that Akt phosphorylation in human endometrium may vary with the menstrual cycle and in early pregnancy and that fluctuations in estrogen level may play a role in Akt activation in endometrial cells. We analyzed Akt phosphorylation using in vivo and in vitro techniques, including Western blot, immunohistochemistry, and immunocytochemistry. Estradiol significantly increased Akt phosphorylation in endometrial cells. Rapid stimulation of Akt activation in cultured stromal cells was observed. Akt phosphorylation by estradiol was inhibited by the PI3K inhibitor, wortmannin, but not by the ER antagonist, ICI 182 780. The maximal effect on Akt activity was observed following 5-15 min of estradiol treatment. Our results suggest that estradiol may directly affect PI3K-related signaling pathway by increasing the phosphorylation of Akt in endometrial cells. Thus, estradiol may exert part of its proliferative and antiapoptotic effects by a nongenomic manner through the Akt signaling pathway.     

Oxidative damage and antioxidant enzyme activities in experimental hypothyroidism

Free radicals are now well known to damage cellular components. To investigate whether age and thyroid level affect peroxidation speed, we examined the levels of malondialdehyde and antioxidant enzyme activities in different age groups of hypothyroid rats. Hypothyroidism was induced in 30- and 60-day-old Wistar Albino rats by the i.p. administration of propylthiouracil (10 mg kg(-1) body weight) for 15 days. While malondialdehyde levels of 30- or 60-day-old hypothyroid rats were increased in liver, they were decreased in the tissues of the heart and thyroid. While glucose-6-phosphate dehydrogenase activity levels did not change in heart, brain and liver tissues of 30-day-old rats, they increased in brain and heart tissues of 60-day-old experimental groups, but decreased in the liver. Catalase activities decreased in the liver and heart of rats with hypothyroidism, but increased in erythrocytes. In control groups while malondialdehyde levels increased in brain, heart and thymus with regard to age, they decreased in plasma. Glucose-6-phosphate dehydrogenase and catalase activities were not affected by age in tissues of the thymus, thyroid and brain, but they were decreased in the heart tissue. The changes in the levels of lipid peroxidation and antioxidant enzyme activities which were determined in different tissues of hypothyroid rats indicate a cause for functional disorder of these tissues. Moreover, there may be changes depending on age at lipid peroxidation and antioxidant enzyme activity levels.     

Col IV and Fn distribution in prostatic adenocarcinoma and correlation of 67LR, MMP-9 and TIMP-1 expression with Gleason score

OBJECTIVE: To assess the immunoreactivity of 5 proteins related to basement membrane (BM) and extracellular matrix in order to investigate whether any of them correlates with differentiation of prostatic adenocarcinoma (PAc). Two of these markers are collagen type IV (Col IV), the collagenous component of basement membrane, and fibronectin (Fn), an adhesion protein in extracellular matrix. Others are matrix metalloproteinase-9 (MMP-9), a type IV collagenase, tissue inhibitor of matrix metalloproteinase-1 (TIMP-1), which has a high affinity for MMP-9, and 67-kd laminin receptor (67LR), which belongs to the non-integrin laminin binding receptor family. STUDY DESIGN: Forty-three PAc cases with Gleason scores ranging between 5 and 10 and 10 benign prostatic hyperplasia (BPH) cases, the control group, were included in the study. Formalin-fixed and paraffin-embedded tissue slides from each case were immunostained with the avidin-biotin-peroxidase method. Immunoreactivity was determined by means of a scoring system similar to the Gleason scoring system. RESULTS: Overexpression of Col IV, Fn, 67LR and MMP-9 was detected in PAc as compared with BPH, whereas no difference was determined in TIMP-1 expression. Among these, only 67LR correlated statistically with Gleason score. CONCLUSION: Expression of 67LR in tumor cells was significantly increased in parallel to tumor grade. This may be useful in microscopic evaluation of PAc.     

Effects of dietary chromium chloride supplementation on performance,some serum parameters,and immune response in broilers

This study was conducted to investigate the effect of increasing dietary levels of inorganic chromium (CrCl₃·6H₂O) on the performance, blood chemistry, and immune response of broilers. Eighty newly hatched Ross PM3 broiler chicks were evenly distributed to five groups of 16 chicks each. Two groups (control and only sheep red blood cell inoculated) were fed the basal diet containing 2.2 and 4.5 mg Cr/kg and the remaining groups were fed 20, 40, or 80 mg/kg Cr-supplemented diets for 44 d. Chicks in all groups, except in the control, at 3 and 5 wk of age, were injected intraperitonally with sheep red blood cell for determining the primary and secondary antibody responses, respectively. When the chicks were 4 wk of age, a delayed-type hypersensitivity test was performed. White blood cells were differentiated. Blood samples were collected for the determination of serum proteins, glucose, cholesterol, cortisol, minerals, and alkaline phosphatase activity and for antibody response. Chromium had no effect on weight gain, but 20 mg/kg supplemental Cr resulted in 18.57% reduction in feed consumption and improved feed efficiency by 16.77%. Chromium did not affect serum cholesterol and P levels but reduced serum glucose and increased serum protein, Cr, Ca, and Mg levels, and ALP activity. A slight reduction was observed with Cr supplementation in cortisol levels. Slight but not significant increases were observed with Cr in serum Zn and Cu. Chromium increased the ratio of bursa of Fabricius and liver to body weight. Heterophil and monocyte counts and heterophil/lymphocyte ratio were reduced and lymphocyte counts, total antibody, IgG, and IgM titers were increased by supplemental Cr. All levels of Cr increased the cell-mediated response to phytohemagglutinin. No alterations in tissues were observed by histopathological examinations.     

Insulin-like growth factor-I and insulin-like growth factor binding protein-3 levels of children living in an iodine- and selenium-deficient endemic goiter area

Serum insulin-like growth factor-I (IGF-I) and insulin-like growth factor binding protein-3 (IGFBP-3) levels were investigated in 31 children living in an endemic goiter area and 33 healthy subjects living in an nonendemic area. Serum IGF-I and IGFBP-3 levels of iodine- and selenium-deficient children were found to be lower than those of control subjects (p<0.001). There was a positive correlation between the IGF-I with chronological age and body mass index. There was also positive correlation between the IGF-I and IGFBP-3. No significant difference was found between the goitrous and nongoitrous children. These results suggest that IGF-I and IGFBP-3 levels are affected by thyroid dysfunction as a result of iodine and selenium deficiency. However, IGF-I and IGFBP-3 levels are not associated with goiter.     

Erythrocyte antioxidant activity and trace element levels in Behçet’s disease

Selenium (Se), zinc (Zn), copper (Cu), and antioxidant enzyme (superoxide dismutase [SOD] and glutathione peroxidase [GSH-Px]) levels in sera were detected in Behçet patients. Age and sex matched controls were used to find out if oxidative stress takes place in the etiopathogenesis of Behçet’s disease. Superoxide dismutase levels were found to be lower in the whole patients group when compared to controls. In whole patients and inactive patients’ group Zn and Se levels were found to be higher, but not different in the active patients group when compared to controls. No significant difference was found between the groups as Cu and glutathione peroxidase levels were taken into consideration. According to the results of the present study, SOD level is low in Behçet’s disease patients’ sera independent from the phase of the disease, and as a result of decreased SOD activity, increased production of free oxygen radicals may play a role in the etiopathogenesis of the disease.     

Ghrelin immunohistochemistry of gastric adenocarcinoma and mucoepidermoid carcinoma of salivary gland.

Ghrelin (G-HH) synthesized in several tissues including salivary and stomach glands stimulates appetite in humans by modulating neuropeptide Y neurons in the hypothalamic arcuate nucleus. Loss of appetite is one of the most important symptoms of stomach cancer. We conducted a study using immunohistochemistry to determine whether salivary glands and stomach cancer tissues produce ghrelin. We determined that negative ghrelin immunohistochemistry discriminates tumors from normal tissues and may therefore further our understanding of the clinically important problem of reduced food intake and anorexia in cancer patients. Radioimmunoassay analyses confirmed that cancer cells do not produce a G-HH peptide, whereas normal cells yield this peptide.     

Metabolic characteristics of experimental free vascularized canine gracilis muscle transfers

A canine gracilis model was used to study muscle energy metabolism and enzyme activities after free vascularized muscle transfer. Fifteen male mongrel dogs underwent orthotopic, free transfer of the left gracilis with microneurovascular anastomosis. After a minimum of 10 months' recovery, muscle biopsy specimens were obtained from the transfers and the contralateral controls and analyzed for relative fiber type areas and maximum activities of phosphorylase, hexokinase, phosphofructokinase, glycerol-3-phosphate dehydrogenase (GPDH), pyruvate kinase, lactate dehydrogenase, citrate synthase, succinate dehydrogenase, 3-hydroxyacyl coenzyme A dehydrogenase (HAD), and creatine phosphokinase. Biopsy specimens obtained before and after a 10 minute, 20-Hz contraction were analyzed for glucose, glycogen, glycolytic intermediates, phosphocreatine, total creatine, and adenine nucleotides (adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, inosine monophosphate, and inosine). There was no significant transfer versus control difference in type I relative fiber area (45 +/- 4 percent versus 44 +/- 3 percent). Total creatine was significantly reduced in the transferred muscles relative to control (83.1 +/- 3.0 mmol/kg versus 100.6 +/- 5.1 mmol/kg dry weight). Maximal activities of phosphorylase, pyruvate kinase, lactate dehydrogenase, citrate synthase, succinate dehydrogenase, HAD, and creatine phosphokinase were diminished in transfers relative to controls, although hexokinase activity was significantly higher in the freely transferred gracilis muscles. During the 20-Hz contraction, muscle transfers produced less force initially, although the force/time integral over the 10-minute stimulation was similar in transfers (277 +/- 25 N/g/second) and controls (272 +/- 24 N/g/second). The contraction was associated with significant glvcogen use and lactate accumulation in both transfers and controls, although this was less pronounced for the transfers. Glycolytic flux appeared muted in the transfers relative to controls. Significant, similar high-energy phosphagen reductions and inosine monophosphate accumulation were noted during the contraction in both groups. Contractile activity is associated with the expected pattern of muscle metabolite changes following free vascularized transfer, indicating the components of cellular energy metabolism are not qualitatively altered after microneurovascular muscle transfer. In contrast, quantitative differences suggest that free vascularized muscle transfer can be associated with a muscle enzyme profile consistent with deconditioning and the presence of denervated muscles fibers in the absence of fiber type profile changes.