Different Concentrations Of Hormones Research Articles

Assessment of DNA methylation changes in tissue culture of Brassica napus.

Plant tissue culture, as a fundamental technique for genetic engineering, has great potential of epigenetic variation, of which DNA methylation is well known of importance to genome activity. We assessed DNA methylation level of explants during tissue culture of Brassica napus (cv. Yangyou 9), using high-performance liquid chromatography (HPLC) assisted quantification. By detecting methylation levels in hypocotyls cultured in mediums with different concentrations of hormones, we found dissected tissue:cultured with 0.1 mg/L 2,4-D and 1.0 mg/L 6-BA, presented the lowest methylation level and highest induction rate of callus (91.0%). Different time point of cultured explants also showed obvious methylation variations, explants cultured after 6 and 21 days exhibited methylation ratios of 4.33 and 8.07%, respectively. Whereas, the methylation ratio raised to 38.7% after 30 days cultivation, indicating that methylation level of hypocotyls ranged during tissue culture. Moreover, we observed that the methylation level in callus is the highest during regeneration of rape-seed, following the regenerated plantlets and hypocotyls. This paper indicated the function of hormones and differentiation of callus is relevant to the methylation levels during tissue culture.

Optimum Protocol for Shoot Formation in karandafi Red Sorghum (<i>Sorghum bicolor</i> (L.) Moench) through Somatic Embryogenesis Using Mature Embryo

In this study, an optimum protocol for shoot formation through somatic embryogenesis using mature embryo explants was developed. Calli were initiated on Murashige and Skoog (MS) media supplemented with varying concentration of 2,4-Dichlorophenoxyl acetic acid (2,4-D) ranging from 1.5 mg/l-4.0 mg/l alone or in combination with 0.5 mg/l Kinetin (KN). Significance difference (p 0.05) was observed among the different concentrations of hormone used for callus induction. The highest percentage callus formation was obtained from the media fortified with 4 mg/l 2, 4-D for mature embryo obtained from imbibed seed while for the preconditioned mature embryo, the media supplemented with 2 m/l 2,4-D + 0.5 mg/l kinetin (KN) recorded more percentage callus formation compared to what was obtained from the media supplemented with 2.5 mg/l2,4-D + 0.5 mg/l. More percentage shoot formation was obtained from the media supplemented with 1 mg/l 6-benzylaminopurine (BA). Average number of shoot per callus was also more in the media fortified with 1 mg/l BA (2.25) but this was not significantly different from what was obtained from the media fortified with 2 mg/l BA+ 0.1 mg/l 2,4-D at 5% level of significance.

Endocrine fingerprint analyses as an alternative strategy to detect the abuse of anabolic steroids

In doping analysis anabolic substances are usually detected by GCMS and LCMS. However, even substances with a chemical structure typical for this class of substances are sometimes not identified when their exact chemical structure is unknown. Therefore, indirect detection techniques may help to complement the classic detection methods in the future. For this purpose the effects of anabolic steroid abuse on the serum concentrations of different hormones were analyzed. Blood and urine samples from a group of male bodybuilders (n = 10) and a male control group (n = 6, 29.7 ± 9.35 years, 81.92 ± 4.18 kg bw, 183 ± 5 cm height) were collected for this study. The urine samples were subsequently analyzed for metabolites of anabolic steroids by the Institute of Biochemistry, German Sport University Cologne. The positively tested bodybuilders were allocated in the group abusing bodybuilders (ABB, n = 5, 24.2 ± 6.3 years, 102 ± 13.77 kg bw, 184 ± 7 cm height) and the negatively tested in the group natural bodybuilders (NB, n = 5, 34.4 ± 14.57 years, 94.5 ± 10.47 kg bw, 182 ± 7 cm height). The serum concentration of cortisol, thyroid-stimulating hormone (TSH), follicle-stimulating hormone (FSH), and luteinizing hormone (LH) in comparison to inhibin, thyroxin (T4), and insulin-like growth factor 1 (IGF-1) was measured by ELISA or RIA. In the ABB T4 and prolactin serum conc. were significantly reduced and the LH conc. significantly enhanced compared to the control group. The TSH serum conc. was significantly reduced in the ABB and NB group compared to the control group. For inhibin a significantly reduced serum conc. in ABB compared to the control group and a tendency toward a difference compared to NB could be observed. The results of this pilot study demonstrate that the comparative analysis of serum concentrations may be suitable to detect doping abuse: TSH as a marker for physical activity; LH, prolactin and inhibin as potential markers for steroid abuse.

Androgen-modulated p21 and p53 gene expression in human non-transformed epithelial prostatic cells in primary cultures

The prostate gland is under androgen control. The aim of the present study was to evaluate the expression of two genes that are regulators of the cell cycle, the p53 and p21 genes, in human non-transformed epithelial prostatic cells (HNTEPs) treated with different concentrations of hormones. Samples of prostate tissue were obtained from 10 patients between 60 and 77 years of age. HNTEP cells were grown in basal medium and treated with dihydrotestosterone (DHT) in different conditions for 4 h. A low concentration of DHT resulted in a significant increase in cell growth; this effect was eradicated by addition of the antiandrogen hydroxyflutamide. Furthermore, the low concentration of DHT induced lower mRNA levels in the p53 and p21 genes in HNTEP cells. In turn, high DHT concentrations induced a significant increase in the expression of the p53 and p21 genes. The present data suggest that the p53 and p21 genes play a role in the control of responsiveness and androgen dose-dependent cell proliferation in HNTEP cells. Further studies are required to assess the intracellular signaling pathway regulated by p53 and p21 under the influence of androgens and its implications for the pathophysiology of prostate diseases.

Effects of media and indole butyric acid (IBA) concentrations on hopbush (Dodoneae viscosa L.) cuttings in green house.

Hopbush (Dodoneae viscosa L.) is an evergreen bush type tree; that is used for hedges and green walls in parks, gardens and houses, in South East of Iran. Propagation by stem cuttings is quicker and cheaper than seed, if the cuttings set in convenient media and rooting hormone. In order to investigate the effects of different media and different concentrations of indole 3-butyric acid (IBA) on rooting of hopbush ( Dodonea viscosa L.), cuttings an experiment was conducted using mist system in greenhouse in spring 2010. The treatments were 3 different media: sand, perlite, and sand + perlite, (1+1 by volume), with 4 levels of IBA concentrations (0 ppm, 2000 ppm, 4000 ppm, and 6000 ppm). A randomized complete block with factorial design was used with 5 replications. The average and the means were compared by Duncun’s multiple range test (1% and 5%); M STAT-C was used for comparing the interaction effects. The effect of medium on number of roots, percentage survival of stem cutting, root fresh weight and dry weight was significant, but on root length was not significantly affected. The effects of different concentrations of hormone on number of roots, percentage survival of stem cutting, root fresh weight and dry weight was significant. The interaction effect of media and hormone on root length was significant, too. The best result was obtained in per-lite, with 4000 ppm IBA. The results showed that perlite medium and 4000 ppm IBA concentration can be suggested for soft wood cutting of hopbush.

Efficient regeneration systems for two closely related Moricandia species possessing a C3 or C3-C4 intermediate photosynthetic character.

The C(3)-C(4) intermediate species Moricandia arvensis ( Brassicaceae) and its closest C(3) relative, Moricandia moricandioides, represent model species for studying the C(3)-C(4) photosynthetic character relative to the C(3) phenotype. In order to enable transgenic analyses in these two species, optimal regeneration systems based on leaf and/or stem internode segments were developed, and genotypes suitable for in vitro tissue culture were identified. Evaluation of the regeneration capability of 30 M. arvensis genotypes and 12 M. moricandioides genotypes revealed that all could form callus. However, shoots were only produced by 40% of the M. arvensis genotypes and 8% of the M. moricandioides genotypes. The two Moricandia species showed significant genotypic differences with respect to callus formation and shoot regeneration. For the 12 regenerative M. arvensis genotypes, 29-100% of the explants developed shoots, while 71% of the explants from the single regenerable M. moricandioides genotype formed shoots. The genotype used, the choice of stem or leaf explants and the composition of the medium (i.e. concentrations of different hormones and salts) significantly affected plant regeneration (chi-square analyses, P<0.05). Whole plants could be obtained in the greenhouse after 3-3.5 months for M. arvensis genotypes and after 4-4.5 months for M. moricandioides.

Changes of hormones and lipoprotein parameters in infants with hypothyroidism; influence of the disease and replacement therapy

Clinical signs of thyroid gland hormones deficiency appear at different ages in infants with different concentrations of hormones. They depend on causes which had caused the hypothyroid degree of subsequents. Concentrations of hormones parameters: tT3, tT4, TSH, fT3, fT4, TBG and Tg (fluoroimmunoassay, DELFIA) and lipid parameters: total cholesterol (Chol), triglycerides (TRG), high density lipoprotein, (HDL-chol) and density lipoprotein, (LDLchol) were determined in order to establish changes that appear under the influence of the disease and replacement therapy in infants with hypothyroidism. In order to establish the influence of infant?s age on a determined parameter, all parameters where determined in the sample of healthy infants. A group of healthy infants (N=100) aged one mounth to 18 years, was devided in 5 age subgroups. The group of patients was devided into the same age groups as healthy group of the moment of diagnosis (N=56). Metabolic condition of an infant was established according to found by ANOVA test according to Tudey Snedecor method for the following parameters; T3 (d=0.35 p&lt;0.001), fT3 (d=1.63 p&lt;0.001) fT4 (d=2.59 p&lt;0.001), TSH (d=2.27 p&lt;0.001), TBG (d=90.82 p&lt;0.001), Tg (d=4.59 p&lt;0.02), Cho. (d=0.48 p&lt;0.001) and LDL-chol. (d=0.51 p&lt;0.001). These findings must be taken into account when the influence of replacement therapy on the observed parameters concentration changes is interpreted. The average concentration values were compared by Student?s t-test in infant before replacement therapy with that of the control subgroup of infants of the same age in order to evaluate the influence on values of the examined parameters. The average concentration values in infants after the replecement therapy were compared that before the replacement therapy as well as the control group, findings in order to the influence on the examined parameters. We observed significant changes hormone parameters in all agesubgroups under hypothyroid condition and return to control levels, except for T4 where the values were significantly higher in all age groups (p&lt;0.05) in comparison to the control group. As for lipids, we found significantly higher values in all age subgroups for chol (p&lt;0.01) and LDL-chol (p&lt;0.001); they did not differ from that in control group after the replacement therapy. Patients were classified at the moment of clinical examination into the following categories: normometabolic (NM) and hypometabolic (HM). They were compared by Student?s T-test of different pairs with the condition before the replacement therapy. The percentage share in the change of TRG concentrations (p&lt;0.02), LDL-chol (p&lt;0.025) and T3 (p&lt;0.001) was significantly different between NM and HM, and the long-lasting replacement therapy caused significant individual differences in Chol concentrations (NM ? 37.90% p&lt;0.001, HM ? 15.14% p&lt;0.001), HDL-chol (NM ? 7.81% p&lt;0.001, HM ? 12.05% p&lt;0.001), LDL-chol (NM ? 50.15% P&lt;0.001 HM 20.03 p&lt;0.01) and T3 (NM ? 105.04% p&lt;0.001, HM 32.18% p&gt;0.005). We conclude that hypothyroidism caused significant hormonal and metabolic changes in infants; however there is no clinical and laboratory index regarding the results of replacement therapy; this is the reason why the patient?s condition must be evaluated individually.

Polycystic ovaries in non-obese and obese patients: possible pathophysiological mechanism based on new interpretation of facts and findings

This study was designed to investigate the basic concentrations of different hormones in obese and non-obese patients with polycystic ovarian disease (PCOD). Eight women with PCOD, of whom four were obese with body mass index (BMI, kg/m2) of > 25 and four were non-obese with BMI < 25, volunteered to participate in this study. Serum samples were taken every 20 min over an 8 h period, starting at 2300 h, on day 5 of a spontaneous or gestagen-induced cycle. Basic insulin concentration was found to be significantly higher in the obese women compared with their non-obese counterparts (P < 0.0001). Serum concentrations of insulin-like growth factor binding protein (IGFBP-I) and sex hormone binding globulin (SHBG) were found to be significantly lower (P < 0.001 for both hormones) in the obese compared with the non-obese women. Serum concentrations of insulin-like growth factor I (IGF-I) did not differ between the two groups. The non-obese women had significantly higher serum concentrations of luteinizing hormone (LH) (P < 0.001) and of growth hormone (GH) (P < 0.002) than their obese counterparts. Based on these results, two models of the development of PCOD were suggested. In obese women, hyperinsulinaemia causes an excessive production of androgens through the enhancement of IGF-I receptors which, in synergism with LH, causes increased activity of cytochrome P-450c 17a. In non-obese patients, relative increase of GH concentration stimulates excessive ovarian IGF-I production. At this point synergism with LH results in excessive production of androgens by the same mechanism as in obese patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Plant regeneration from cucumber ( Cucumis sativus) protoplasts

A procedure is described for the isolation and culture of protoplasts from cotyledons and leaves of Cucumis sativus. The plants used for protoplast isolation were grown in vitro. Protoplasts were plated according to different procedures, which were compared on the basis of the observed plating efficiency. Protoplasts cultured in agarose solidified medium (either by the bead-type culture or the disc-type culture) showed plating efficiencies of up to 50%. In contrast, division frequency was low (<2%) when protoplasts were cultured in liquid medium. Frequent removal of the medium and addition of fresh medium with lower osmolarity and different concentrations of hormones resulted in the production of yellow capable capable of regeneration. Shoots were obtained from leaf protoplasts approx. 3 months after protoplast isolation. It was possible to induce regeneration in calli subcultured for 1 year.

Influence of hydrocortisone on human fetal small intestine in organ culture.

The influence of hydrocortisone on the differentiation and proliferation of human fetal small intestine was studied. Fetal intestine (12- to 14-week gestation) was cultured during 5 days at 37 degrees C in serum-free Leibovitz L-15 medium alone or supplemented with hydrocortisone (12.5, 25, and 50 ng/ml). The addition of different concentrations of hormone did not affect the morphology of the intestinal explants. Brush border membrane hydrolytic activities, namely, sucrase, lactase, glucoamylase, trehalase, and alkaline phosphatase activities, were assayed in the intestinal tissue. A specific increase of lactase and alkaline phosphatase activities was induced by the addition of 25 and 50 ng hydrocortisone/ml culture medium. The DNA synthesis evaluated by the incorporation of [3H]thymidine was increased by the addition of 50 ng hydrocortisone/ml. The sites of incorporation into the different layers of the intestinal wall were studied by radioautography. The incorporation of the radioactive precursor occurred mainly in the epithelium and to a lesser degree in the mesenchyme and muscular layers. Labeled epithelial nuclei were located in the intervillous areas and developing crypts but not on the villi. The addition of hydrocortisone induced a significant increase of the labeling index of the epithelial cells. The present work provided for the first time some basic data on the influence of hydrocortisone on brush border hydrolytic activities and on epithelial cell proliferation of human fetal small intestine.

The success rate of in vitro fertilization of human oocytes in relation to the concentrations of different hormones in follicular fluid and peripheral plasma

Seventeen infertile patients were stimulated with clomiphene citrate and human chorionic gonadotrophin (hCG). During laparotomy or laparoscopy, follicles were aspirated. At least one fertilizable oocyte was obtained from 13 patients, who had normal preovulatory plasma 17 beta-estradiol (mean 2.2 nmol/l) in contrast to the 4 patients with infertilizable oocytes (mean 0.4 nmol/l). A close association was found between the success rate of fertilization and the increment of plasma progesterone from the day before to the day after the operation. If the concentrations of 17 beta-estradiol and progesterone in follicular fluid were below an arbitrary limit (17 beta-estradiol less than 500 nmol/l in association with progesterone less than 2,000 nmol/l), no oocytes could be fertilized. Apparently aspiration of oocytes and follicular fluid did not disturb ovarian steroid production during the luteal phase. The results suggest that plasma 17 beta-estradiol and progesterone determinations may serve as useful predictors for oocyte fertilizability in an in vitro system, following clomiphene-hCG stimulation and multiple oocyte recovery.

The effect of hormones on the haemodynamics in animals of different age groups

The investigation was undertaken to study to effect of varying doses of epinephrine, vasopressin, insulin, thyroxin, estradiol-dipropionate on the haemodynamics and contractility of the myocardium of rats in different age groups, as well as the effect of epinephrine and vasopressin on coronary blood circulation in dogs of various age groups. The age group peculiarities of reactions to the hormones were compared with changes in their content in the blood of animals of different age groups. Upon administering small concentrations of the hormones in this study, more pronounced changes in haemodynamics and contractility of the myocardium were observed in senile animals. The administration of larger doses led to more pronounced changes in mature animals. During the process of aging, there is a decrease in the range of the reactions of the cardiovascular system to the action of hormones. In case of long term (14 days) administration of thyroxin, it was found that in senile animals the contractility of the heart was lower than that in short term (4 days) administration. In long term administration of thyroxin, it was found that in mature animals the index for the myocardium's contracting ability was more pronounced than in short term administration. Irregular changes in the concentrations of different hormones in the blood, changes in sensitivity and ability of the myocardium to respond to their action lead to changes in the hormonal regulation of the heart, and to the development of aging metabolic and functional changes.

Renal and peripheral vascular responsiveness to arginine vasotocin in bullfrog, Rana catesbeiana.

Arginine vasotocin (AVT), the amphibian neurohypophysial hormone, has pressor and antidiuretic actions in bullfrogs (Rana catesbeiana). Much higher doses of exogenous AVT were required for the pressor response than those for the antidiuretic response. When bullfrog legs and kidneys were perfused under constant pressure simultaneously, the kidney vasculature was more sensitive to bolus injection of AVT than was the leg vasculature. The same was true when the tissues were perfused with different concentrations of hormone. AVT constricts preglomerular vessels in the bullfrog. We suggest that the preglomerular vasculature of the bullfrog is more sensitive to AVT than is the peripheral vasculature. This may explain in part the antidiuretic action of the hormone at nonpressor doses. Evolutionary speculations are discussed.

Experimental analysis of development of cardiac insufficiency in old age

The investigation was undertaken to study to effect of varying doses of epinephrine, vasopressin, insulin, thyroxin, estradiol-dipropionate on the haemodynamics and contractility of the myocardium of rats in different age groups, as well as the effect of epinephrine and vasopressin on coronary blood circulation in dogs of various age groups. The age group peculiarities of reactions to the hormones were compared with changes in their content in the blood of animals of different age groups.Upon administering small concentrations of the hormones in this study, more pronounced changes in haemodynamics and contractility of the myocardium were observed in senile animals. The administration of larger doses led to more pronounced changes in mature animals. During the process of aging, there is a decrease in the range of the reactions of the cardiovascular system to the action of hormones.In case of long term (14 days) administration of thyroxin, it was found that in senile animals the contractility of the heart was lower than that in short term (4 days) administration. In long term administration of thyroxin, it was found that in mature animals the index for the myocardium's contracting ability was more pronounced than in short term administration.Irregular changes in the concentrations of different hormones in the blood, changes in sensitivity and ability of the myocardium to respond to their action lead to changes in the hormonal regulation of the heart, and to the development of aging metabolic and functional changes.

The plasmatic release and adrenal storage of adrenaline and noradrenaline during third part of pregnancy, parturition and post-partum in the rat

The evolution of adrenaline and noradrenaline in plasma and adrenals of rats during pregnancy, parturition and post parturition was studied. Plasmatic adrenaline demonstrated marked increases on the 18th day of pregnancy and during parturition. At the same time adrenaline stores of adrenal gland declined. Noradrenaline in plasma was significantly lower during 16th to 20th day of pregnancy and during parturition. The noradrenaline storage in adrenal gland followed nearly similar pattern as in the plasma. An accelerated rate of adrenaline release in the blood from adrenal gland during pregnancy is suggested. Noradrenaline variations appear to be the consequence of changed sympathetic nervous activity due to modifications in the endocrine status of the pregnant rats. The results suggest that variations in the concentrations of different hormones induced by the pregnancy produce altered pattern of adrenaline and noradrenaline release and storage.

Effects of individual or combined administration of reserpine and niamide on plasmatic release and adrenal storage of catecholamines in pregnant rats

Summary Plasmatic adrenaline and noradrenaline in 20 days pregnant rats were markedly lower than the controls. Adrenal stores of noradrenaline during pregnancy were higher than controls but adrenal adrenaline in pregnant rats was lower. The effects of reserpine and niamide administration in control and pregnant rats differed in the intensities of their actions and showed high statistical significances. 24 hours after a single injection of reserpine, adrenaline in plasma and adrenals of control rats declined. Similar treatment to pregnant rats declined plasmatic adrenaline but both adrenal and plasmatic noradrenaline increased. Niamide administration reduced adrenal and plasmatic adrenaline in controls but in pregnants adrenaline in plasma increased and decreased in adrenals. Adrenal stores of noradrenaline in both cases were augmented after niamide administration. There was a high rise in plasmatic noradrenaline in pregnant rats after niamide injection but in controls it declined. The combined administration of reserpine and niamide depressed plasmatic adrenaline in control as well as pregnant rats. Plasmatic noradrenaline in controls reduced with rise in adrenal stores but the pregnant rats demonstrated increased plasmatic noradrenaline and decreased adrenal noradrenaline after combined administration. The differences in intensities and modes of actions of both drugs in control and pregnant rats appear to be the consequence of hormonal state of rats and suggest that variable concentrations of different hormones in pregnancy counteract the effects of reserpine and niamide.