Alterations induced by juvenile hormone in glandular cells of the Apis mellifera venom gland I - Application on the larvae (Hymenoptera: Apidae)

Histological analyses were made in order to evaluate the effects of the topic application of a synthetic juvenile hormone (JH-III Sigma) on the development of the venom glands in workers of Apis mellifera. Three experimental groups were used: the first received 1 μl of a dilution of the juvenile hormone in hexane (2μg/μl); the second group received 1 μl of hexane; and the third group, the control, did not receive any kind of treatment. The application was made on larvae at the beginning of the fifth instar and the glands were collected at different developmental stages. The results showed that the application of the diluted hormone, as well as the hexane alone, accelerated gland development in relation to the control group at all developmental stages studied. These data suggest that the juvenile hormone acts on the development of the venom gland; nevertheless, this action could be amplified by the effect of the solvent used in the present work, as well as in other studies concerning this matter.

Juvenile hormone promotes changes in the expression of hypopharyngeal gland proteins of worker Apis mellifera (Hymenoptera: Apidae)

The present investigation compares the protein electrophoreses profiles of the hypopharyngeal glands of 12 and 25 day old Apis mellifera workers, some of which were experimentally treated with an analogue of juvenile hormone in the moment of the emergence while others were not treated. According to the evaluation of the presented variations by four main bands, it is concluded that the analogue juvenile hormone changes the glandular genetic expression pattern, promoting the disappearance of two from the four main bands in 25 day old workers. The effect of this hormone is discussed as an hypopharyngeal maturation inductor, in synergetic action with the bee age acting early in the glandular cycle.

Influence of the application of juvenile hormone on the haemolymph total proteins and electrophoretic pattern of worker larvae of Apis mellifera

The aim of the present work was to verify the influence of the juvenile hormone (JH) applied on worker larvae of Apis mellifera 2 to 5 days old over the haemolymph total protein and electrophoretic pattern. Each larvae received topical applications of 1 ml of a solution of JH in hexane (1 μg/ml) on their 2 nd, 3 rd 4 th and 5 th day after hatching and had the amount and electrophoretic pattern of proteins from the haemolymph analyzed during the remaining days of their life. As a control, haemolymph of larvae of the same age that did not receive any kind of treatment was analyzed. The results show that the application of JH on larvae 3 or more days old affect the amount and electrophoretic pattern of the proteins, with this effect lasting through the subsequent days.

Superovulation in Cycling Mares Using Equine Follicle Stimulating Hormone (eFSH)

Superovulation would potentially increase the efficiency and decrease the cost of embryo transfer by increasing embryo collection rates. Other potential clinical applications include improving pregnancy rates from frozen semen, treatment of subfertility in stallions and mares, and induction of ovulation in transitional mares. The objective of this study was to evaluate the efficacy of purified equine follicle stimulating hormone (eFSH; Bioniche Animal Health USA, Inc., Athens, GA) in inducing superovulation in cycling mares. In the first experiment, 49 normal, cycling mares were used in a study at Colorado State University. Mares were assigned to 1 of 3 groups: group 1, controls (n = 29) and groups 2 and 3, eFSH-treated (n = 10/group). Treated mares were administered 25 mg of eFSH twice daily beginning 5 or 6 days after ovulation (group 2). Mares received 250 (of cloprostenol on the second day of eFSH treatment. Administration of eFSH continued until the majority of follicles reached a diameter of 35 mm, at which time a deslorelin implant was administered. Group 3 mares (n = 10) received 12 mg of eFSH twice daily starting on day 5 or 6. The treatment regimen was identical to that of group 2. Mares in all 3 groups were bred with semen from 1 of 4 stallions. Pregnancy status was determined at 14 to 16 days after ovulation. In experiment 2, 16 light-horse mares were used during the physiologic breeding season in Brazil. On the first cycle, mares served as controls, and on the second cycle, mares were administered 12 mg of eFSH twice daily until a majority of follicles were 35 mm in diameter, at which time human chorionic gonadotropin (hCG) was administered. Mares were inseminated on both cycles, and embryo collection attempts were performed 7 or 8 days after ovulation. Mares treated with 25 mg of eFSH developed a greater number of follicles (35 mm) and ovulated a greater number of follicles than control mares. However, the number of pregnancies obtained per mare was not different between control mares and those receiving 25 mg of eFSH twice daily. Mares treated with 12 mg of eFSH and administered either hCG or deslorelin also developed more follicles than untreated controls. Mares receiving eFSH followed by hCG ovulated a greater number of follicles than control mares, whereas the number of ovulations from mares receiving eFSH followed by deslorelin was similar to that of control mares. Pregnancy rate for mares induced to ovulate with hCG was higher than that of control mares, whereas the pregnancy rate for eFSH-treated mares induced to ovulate with deslorelin did not differ from that of the controls. Overall, 80% of mares administered eFSH had multiple ovulations compared with 10.3% of the control mares. In experiment 2, the number of large follicles was greater in the eFSH-treated cycle than the previous untreated cycle. In addition, the number of ovulations during the cycle in which mares were treated with eFSH was greater (3.6) than for the control cycle (1.0). The average number of embryos recovered per mare for the eFSH cycle (1.9 ± 0.3) was greater than the embryo recovery rate for the control cycle (0.5 ± 0.3). In summary, the highest ovulation and the highest pregnancy and embryo recovery rates were obtained after administration of 12 mg of eFSH twice daily followed by 2500 IU of hCG. Superovulation with eFSH increased pregnancy rate and embryo recovery rate and, thus, the efficiency of the embryo transfer program.

Electrophoretical Protein Profile in Apis mellifera (Hymenoptera; Apidae) Hypopharyngeal Glands under Juvenile Hormone Effect When the Colony is Grown in the Absence of Brood

Twelve-day-old and 25-day-old Apis mellifera workers were treated or not treated with juvenile hormone at the moment of emergence and reared in the colony without brood. Having the brood interference apart, the hormone effect on the hypopharyngeal glands protein expression was determined through the electrophoretical protein profiles of the both groups of bees. In those conditions, the hormone induced changes that were different from the control. Protein bands of 66 and 48 kDa were intensified in the 12-day-old bees, whereas band of 42 kDa was reduced in the 25-day-old bees. That indicated a different effect of the juvenile hormone in the function of bee aging, which promoted a glandular protein activation in the young bees and, in contrast, an inhibitory action in the 25-day-old bees workers.

Juvenile Hormone Effect on the Venom Gland Secretory Cycle in Workers of Apis mellifera (Hymenoptera, Apidae)

The present investigation analyzed the influence of Juvenile Hormone (JH) on the venom glands of Apis mellifera workers through protein dosage and electrophoresis of venom gland extracts of newly emerged workers which were treated with 1 μl JH dissolved in hexane, in concentration of 2μg/μl. Newly emerged workers non-treated and treated with 1 μl hexane were the controls. Both JH and hexane provoke quantitative changes on the gland protein titre and on the protein electrophoretic profile. The disappearance of protein bands in the venom gland extracts of 14 day-old treated workers, a situation normally found only in 35 day-old non-treated workers, suggests that the JH treatment induces a precocious maturation of the worker venom gland.

Diagnosis of 5α-reductase type 2 deficiency: Contribution of anti-Müllerian hormone evaluation

Aim: To evaluate anti-Müllerian hormone (AMH) levels in patients with clinical and molecular diagnosis of 5α-reductase 2 deficiency. Patients and methods: Data from 14 patients whose age ranged from 21 days to 29 years were analyzed according to age and pubertal stage. Sexual ambiguity was rated as Prader III in 11 patients. LH, FSH, testosterone (T), dihydrotestosterone (DHT) and AMH serum levels were measured in all but two patients, who had been previously submitted to gonadectomy; T and DHT were also measured in 20 age-matched controls. Results: Gonadotropin levels were normal in all but one patient who retained gonads (six of whom had reached puberty) and T/DHT ratio was elevated in all patients when compared to controls. All prepubertal patients had AMH levels < -1 SD for age, while most pubertal patients had AMH levels compatible with pubertal stage. Conclusions: Prepubertal patients with 5α-reductase 2 deficiency have AMH values in the lower part of the normal range. These data indicate that T does not need to be converted to DHT to inhibit AMH secretion by Sertoli cells. © Freund Publishing House Ltd., London.

Efficacy of hormone therapy with and without methyltestosterone augmentation of venlafaxine in the treatment of postmenopausal depression: A double-blind controlled pilot study

Objective: This study evaluated the augmentation of venlafaxine with hormone therapy in the treatment of postmenopausal depression. The hormones evaluated were estrogen (0.625 mg) in combination with medroxyprogesterone acetate (2.5 mg) and methyltestosterone (2.5 mg). Design: Seventy-two menopausal women (mean age: 53.6 ± 4.27 years) diagnosed with depression (Montgomery-Åsberg Depression Rating Scale [MADRS] scores ≥ 20) were treated with venlafaxine and one of the following hormone therapy combinations, in a double-blind regimen: estrogen + medroxyprogesterone + methyltestosterone (group 1, n = 20); estrogen + medroxyprogesterone acetate (group 2, n = 20); methyltestosterone only (group 3, n = 16); and no hormone therapy (group 4, n = 16). Study duration was 24 weeks. Primary efficacy outcome was remission according to the MADRS, whereas secondary efficacy measures included the Clinical Global Impression (CGI), Blatt-Kupperman Index, and Women's Health Questionnaire (WHQ). Results: Forty-eight patients completed the study. All groups showed significant improvement from baseline. Group 3 demonstrated significant improvement on the MADRS compared with placebo (group 4) at weeks 20 (P = 0.048) and 24 (P = 0.030); effect size 8.04 (0.83; 15.26) (P = 0.029), but also had the highest dropout rate. Groups 1 and 3 had significant CGI improvement rates compared with placebo: 42.23% (P = 0.012) and 44.45% (P = 0.08), respectively. There were no differences in the WHQ or BKI scores among the groups. Conclusions: Methyltestosterone 2.5 mg had the highest effect size compared with placebo, but the high dropout rate prevented its efficacy from being determined. Estrogen plus medroxyprogesterone, combined with methyltestosterone or otherwise, demonstrated a trend toward increased efficacy of venlafaxine. Further larger-scale clinical trials are needed to elucidate the findings of this pilot study. © 2006 by The North American Menopause Society.

Substitution-newton-Raphson method for the solution of electric network equations

The power flow problem, in transmission networks, has been well solved, for most cases, using Newton-Raphson method (NR) and its decoupled versions. Generally speaking, the solution of a non-linear system of equations refers to two methods: NR and Successive Substitution. The proposal of this paper is to evaluate the potential of the Substitution-Newton-Raphson Method (SNR), which combines both methods, on the solution of the power flow problem. Simulations were performed using a two-bus test network in order to observe the characteristics of these methods. It was verified that the NR is faster than SNR, in terms of convergence, considering non-stressed scenarios. For those cases where the power flow in the network is closed to the limits (stressed system), the SNR converges faster. This paper presents the power flow formulation of the SNR and describes its potential for its application in special cases such as stressed scenarios. © 2006 IEEE.

The importance of hormone receptor analysis in osteosarcoma cells growth submitted to treatment with estrogen in association with thyroid hormone

Bone tumor incidence in women peaks at age 50-60, coinciding with the menopause. That estrogen (E2) and triiodothyronine (T3) interact in bone metabolism has been well established. However, few data on the action of these hormones are available. Our purpose was to determine the role of E2 and T3 in the expression of bone activity markers, namely alkaline phosphatase (AP) and receptor activator of nuclear factor κB ligand (RANKL). Two osteosarcoma cell lines: MG-63 (which has both estrogen (ER) and thyroid hormone (TR) receptors) and SaOs-29 (ER receptors only) were treated with infraphysiological E2 associated with T3 at infraphysiological, physiological, and supraphysiological concentrations. Real-time RT-PCR was used for expression analysis. Our results show that, in MG-63 cells, infraphysiological E2 associated with supraphysiological T3 increases AP expression and decreases RANKL expression, while infraphysiological E2 associated with either physiological or supraphysiological T3 decreases both AP and RANKL expression. On the other hand, in SaOs-2 cells, the same hormone combinations had no significant effect on the markers' expression. Thus, the analysis of hormone receptors was shown to be crucial for the assessment of tumor potential growth in the face of hormonal changes. Special care should be provided to patients with T3 and E2 hormone receptors that may increase tumor growth. Copyright © 2007 John Wiley & Sons, Ltd.

Morphologic changes and the expression of alpha-melanocyte stimulating hormone and melanocortin-1 receptor in melasma lesions: A comparative study

Melasma is a common acquired symmetrical hypermelanosis characterized by irregular light- to dark-brown macules on sun-exposed skin areas. The literature shows few studies on its physiopathogeny. However, changes in α-melanocyte stimulating hormone (α-MSH) secretion and melanocortin-1 receptor (MC1-R) expression may play a role to trigger this condition. Biopsies were taken from both melasma skin and adjacent perilesional normal skin of 44 patients. The biopsies were submitted for hematoxylin and eosin and Fontana-Masson staining and immunohistochemistry with Melan-A, α-MSH, and MC1-R, and processed for transmission electron microscopy. In some cases, they were submitted to MC1-R gene expression analysis by real-time polymerase chain reaction. Increased lymphohistiocytic infiltrate and solar elastosis, higher epidermal melanin were observed in melasma skin. Electron microscopy revealed a greater number of mature melanosomes in keratinocytes and melanocytes, and more prominent cytoplasmic organelles in melasma skin. There was no difference in melanocyte number between areas. However, melanocytes were larger and more dendritic in melasma skin. Immunohistochemistry with α-MSH and MC1-R showed significant labeling in melasmic epidermis but MC1-R messenger ribonucleic acid (RNAm) did not show significant quantitative difference between melasma and normal skin. © 2010 by Lippincott Williams & Wilkins.

Analysis of agonist and antagonist effects on thyroid hormone receptor conformation by hydrogen/deuterium exchange

Thyroid hormone receptors (TRs) are ligand-gated transcription factors with critical roles in development and metabolism. Although x-ray structures of TR ligand-binding domains (LBDs) with agonists are available, comparable structures without ligand (apo-TR) or with antagonists are not. It remains important to understand apo-LBD conformation and the way that it rearranges with ligands to develop better TR pharmaceuticals. In this study, we conducted hydrogen/deuterium exchange on TR LBDs with or without agonist (T 3) or antagonist (NH3). Both ligands reduce deuterium incorporation into LBD amide hydrogens, implying tighter overall folding of the domain. As predicted, mass spectroscopic analysis of individual proteolytic peptides after hydrogen/ deuterium exchange reveals that ligand increases the degree of solvent protection of regions close to the buried ligand-binding pocket. However, there is also extensive ligand protection of other regions, including the dimer surface at H10-H11, providing evidence for allosteric communication between the ligand-binding pocket and distant interaction surfaces. Surprisingly, Cterminal activation helix H12, which is known to alter position with ligand, remains relatively protected from solvent in all conditions suggesting that it is packed against the LBD irrespective of the presence or type of ligand. T 3, but not NH3, increases accessibility of the upper part of H3-H5 to solvent, and we propose that TR H12 interacts with this region in apo-TR and that this interaction is blocked by T 3 but not NH3.Wepresent data from site-directed mutagenesis experiments and molecular dynamics simulations that lend support to this structural model of apo-TR and its ligand-dependent conformational changes. (Molecular Endocrinology 25: 15-31, 2011). Copyright © 2011 by The Endocrine Society.

Adjuvant therapy with GnRH agonists/tamoxifen in breast cancer should be a good council for patients with hormone receptor-positive tumours and wish to preserve fertility

Infertility represents one of the main long-term consequences of combination chemotherapy used for the treatment of breast cancer. Approximately 60%-65% of breast cancers express the nuclear hormone receptor in premenopausal women. Adjuvant endocrine therapy is an integral component of care for patients with hormone receptor-positive (HR+) tumours. The GnRH agonist (GnRHa) alone or in combination with tamoxifen produces results at least similar to those obtained with the different chemotherapy protocols in patients with HR+ tumors with respect to recurrence-free survival and overall survival, Presentation of the hypothesis: It is time to indicate adjuvant therapy with GnRHa associated with tamoxifen for patients with breast cancer (HR+ tumours) if they want to preserve their reproductive function. Testing the hypothesis: Assessment of ovarian reserve tests: follicle stimulating hormone (FSH), anti-Mullerian hormone (AMH), inhibin B, antral follicle count (AFC) and ovarian volume 6 months, and 1 year after the end of therapy with GnRHa/tamoxifen. The recurrence-free survival and overall survival should be analysed. Implications of the hypothesis: The major implication will be to avoid adjuvant chemotherapy for patients with breast cancer (HR+ tumours) that request fertility preservation. It is expected that ovarian function should not be altered in almost all cases. © Todos os direitos reservados a SBRA - Sociedade Brasileira de Reprodução Assistida.

Substitution of corn and soybean oil by cassava meal and African palm tree oil in diets of laying hens

Fourty-two White Leghorns laying hens, from the commercial Cuban hybrid L-33, were used for eight weeks during the laying peak (36 to 43 weeks of age), to assess the substitution of corn by cassava root meal (Manihot esculenta Crantz) and the crude soybean oil by crude oil of African palm tree (Elaeis guineensis J.) in the diets of laying hens. Analysis of variance was conducted, according to simple classification design, with three treatments and 14 repetitions (a cage with a hen). The treatments consisted of three diets (1- corn meal + soybean oil; 2- 25 % cassava meal + African palm tree oil; 3- 53 % cassava meal + African palm tree oil), with 15.71 % CP; 3.83 % Ca and 0.36 % P available. The viability was of 100 % in all treatments. No differences were found for laying (92.21, 92.09 and 91.59 %), which surpassed the potential of this hybrid during the laying peak (90 %), conversion (118g feedstuff/egg in the three treatments), egg mass produced (3066, 3114 and 3071 g/bird) and mass conversion (1.99, 1.95 y 1.98 feed consumed/egg mass). The pigmentation of the egg yolk was reduced as the level of cassava meal increased in the diets (6, 4 and 3 at Roche's scale), as well as the cost of the feed consumed in 56 d per hen (2.56, 2.15 and 1.83 USD/bird). The possibility of substituting, totally, corn meal by that of cassava and soybean oil by that of the African palm tree in the diets of laying hens during the laying peak was determined, with positive economic effect and without damaging the productive performance of birds.