Roles of estrogen and progesterone in modulating renal nerve function in the rat kidney

The maintenance of extracellular Na+ and Cl- concentrations in mammals depends, at least in part, on renal function. It has been shown that neural and endocrine mechanisms regulate extracellular fluid volume and transport of electrolytes along nephrons. Studies of sex hormones and renal nerves suggested that sex hormones modulate renal function, although this relationship is not well understood in the kidney. To better understand the role of these hormones on the effects that renal nerves have on Na+ and Cl- reabsorption, we studied the effects of renal denervation and oophorectomy in female rats. Oophorectomized (OVX) rats received 17β-estradiol benzoate (OVE, 2.0 mg·kg-1·day-1, sc) and progesterone (OVP, 1.7 mg·kg-1·day-1,sc). We assessed Na+ and Cl-fractional excretion (FENa+ and FECl-, respectively) and renal and plasma catecholamine release concentrations. FENa+, FECl-, water intake, urinary flow, and renal and plasma catecholamine release levels increased in OVX vs control rats. These effects were reversed by 17β-estradiol benzoate but not by progesterone. Renal denervation did not alter FENa+, FECl-, water intake, or urinary flow values vs controls. However, the renal catecholamine release level was decreased in the OVP (236.6±36.1 ng/g) and denervated rat groups (D: 102.1±15.7; ODE: 108.7±23.2; ODP: 101.1±22.1 ng/g). Furthermore, combining OVX + D (OD: 111.9±25.4) decreased renal catecholamine release levels compared to either treatment alone. OVE normalized and OVP reduced renal catecholamine release levels, and the effects on plasma catecholamine release levels were reversed by ODE and ODP replacement in OD. These data suggest that progesterone may influence catecholamine release levels by renal innervation and that there are complex interactions among renal nerves, estrogen, and progesterone in the modulation of renal function.

Behavioral effects of endogenous or exogenous estradiol and progesterone on cocaine sensitization in female rats

Cocaine sensitization is a marker for some facets of addiction, is greater in female rats, and may be influenced by their sex hormones. We compared the modulatory effects of endogenous or exogenous estradiol and progesterone on cocaine-induced behavioral sensitization in 106 female rats. Ovariectomized female rats received progesterone (0.5 mg/mL), estradiol (0.05 mg/mL), progesterone plus estradiol, or the oil vehicle. Sham-operated control females received oil. Control and acute subgroups received injections of saline, while the repeated group received cocaine (15 mg/kg, ip) for 8 days. After 10 days, the acute and repeated groups received a challenge dose of cocaine, after which locomotion and stereotypy were monitored. The estrous cycle phase was evaluated and blood was collected to verify hormone levels. Repeated cocaine treatment induced overall behavioral sensitization in female rats, with increased locomotion and stereotypies. In detailed analysis, ovariectomized rats showed no locomotor sensitization; however, the sensitization of stereotypies was maintained. Only females with endogenous estradiol and progesterone demonstrated increased locomotor activity after cocaine challenge. Estradiol replacement enhanced stereotyped behaviors after repeated cocaine administration. Cocaine sensitization of stereotyped behaviors in female rats was reduced after progesterone replacement, either alone or concomitant with estradiol. The behavioral responses (locomotion and stereotypy) to cocaine were affected differently, depending on whether the female hormones were of an endogenous or exogenous origin. Therefore, hormonal cycling appears to be an important factor in the sensitization of females. Although estradiol increases the risk of cocaine sensitization, progesterone warrants further study as a pharmacological treatment in the prevention of psychostimulant abuse.

Evaluación invitro e in vivo de las características de liberación de gabapentina a partir de biomateriales obtenidos vía sol-gel.

Tesis (Maestría en Ciencias con Orientación en Farmacia) UANL, 2011.

Activation of Progesterone Receptor by ATP

Progesterone-receptor complex from freshly prepared hen oviduct cytosol acquired the ability to bind to isolated nuclei, DNA-cellulose and ATP-Sepharose when incubated with 5-10 mM ATP at 4°C. The extent of this ATP-dependent activation was higher when compared with heat-activation achieved by warming the progesterone- receptor complex at 23 °C. The transformation of progesterone-receptor complex which occurred in a time-dependent manner was only partially dependent on hormone presence. The ATP effect was selective in causing this transformation whereas ADP, AMP and cAMP failed to show any such effect. The non-hydrolizable analogs of ATP, adenosine 5'-[a,/3-methylene]triphosphate and adenosine 5-[/l,y-imido]triphosphate were also found to be ineffective. Presence of 10 mM sodium molybdate blocked both the ATP and the heat-activation of progesterone-receptor complex. Mn" or Mg` had no detectable effect on the receptor activation but the presence of Ca" increased the extent of ATP-activation slightly. EDTA presence (> 5 mM) decreased the extent of receptor activation by about 40 % and was, therefore, not included in the buffers used for activation studies. Divalent cations were also ineffective when tested in the presence of 1- 5 mM EDTA. The properties of progesterone-receptor complex remained intact under the above conditions when analyzed for steroid-binding specificity and Scatchard analysis. However, the ATP-activated progesterone-receptor complex lost the ability to aggregate when tested on low-salt sucrose gradients. ATP was equally effective in activating the rat-uterine estradiol-receptor complex at 4 "C and influenced the transformation of 4-S receptor form into a 5-S form when analyzed on sucrose gradients containing 0.3 M KCI. The presence of ATP also increased the rate of activation of progesterone-receptor complex at 23 °C. These findings suggest a role for ATP in receptor function and offer a convenient method of studying the process of receptor activation at low temperature and mild assay conditions.

Metabolic profiles and progesterone cycles in first lactation dairy cows

This study investigated the ovarian function, metabolic profiles and fertility in first lactation Holstein-Friesian dairy cows (mean 305 day milk yield: 7417 +/- 191 kg, n = 37). Reproductive profiles obtained from milk progesterone analysis were categorized into normal (n = 17) and four abnormal profiles (delayed ovulation, DOV1, n = 9; DOV2, n = 2; persistent corpus luteum, PCL1, n = 6; PCL2, n = 4; 1: immediately post-calving, 2: subsequent cycles). Fifty-five percent of cows had abnormal profiles with half of these being categorized as DOV1. Fertility of DOV1 and DOV2 cows was reduced whereas PCL1 and PCL2 cows had similar reproductive competence to normal profile cows. DOV1 animals had higher milk energy values, lower energy balances, lower dry matter intakes (DMI) and greater body weight and body condition score (BCS) losses post-calving than normal profile animals. DOV1 animals also had lower insulin-like growth factor-I (IGF-I) and higher betahydroxybutyrate (BHB) concentrations and tended to have the lower insulin and glucose concentrations in the pre-service period than normal profile cows. All PCL animals had vulval discharges postpartum. Despite this, the DMI, body weight and BCS changes, IGF-I concentrations and fertility of PCL1 animals was similar to normal profile cows. In conclusion, the high prevalence of delayed ovulation post-calving (DOV1) in primiparous high yielding cows lasted long enough (71 +/- 8.3 days) to have a detrimental impact on fertility and was associated with significant physiological changes. This study did not establish any detrimental effects of PCL profiles on fertility or production parameters. (C) 2002 Elsevier Science Inc. All rights reserved.

Bone morphogenetic protein-6 enhances gonadotrophin-dependent progesterone and inhibin secretion and expression of rnRNA transcripts encoding gonadotrophin receptors and inhibin/activin subunits in chicken granulosa cells

The aims were to examine ovarian expression of bone morphogenetic protein (BMP) ligands/receptor mRNAs in the chicken and to test the hypothesis that theca-derived BMP(s) modulates granulosa cell function in a paracrine manner. RT-PCR revealed expression of multiple BMPs in granulosa and theca cells from prehierarchical and preovulatory follicles with greater expression in theca cells; both cell types expressed BMP receptors-1A, -1B and -II consistent with tissue responsiveness. Preovulatory granulosa cells F1, F2 and F3/4) were cultured with BMP-6 (expressed by theca but not granulosa) in the presence/absence of LH, FSH or 8-Br-cAMP. RMP-6 increased 'basal' and gonadotrophin-induced inhibin-A and progesterone secretion by each cell type but did not enhance the effect of 8-Br-cAMP. This indicates that the observed synergism between BMP-6 and gonadotrophin might involve BMP-induced up-regulation of gonadotrophin receptors. In support of this, BMP-6 alone increased LH-receptor (LHR) mRNA in F1 cells and FSH-receptor (FSHR) mRNA in F1, F2 and F3/4 cells. RMP-6 also enhanced LH/FSH-induced LHR transcript amount in each cell type but did not raise FSHR transcript amounts above those induced by BMP-6 alone. To further explore BMP6 action on inhibin-A secretion, we quantified inhibin/activin subunits (alpha, beta(A), beta(B)) mRNAs. Consistent with its effect on inhibin-A secretion, BMP-6 enhanced 'basal' expression of alpha- and beta(A)-Subunit mRNA in F1, F2 and F3/4 cells, and beta(B)-subunit mRNA in F3/4 cells. BMP-6 markedly enhanced FSH/LH-induced expression of alpha-subunit in all follicles and FSH-induced beta(A)-subunit in F2 and F3/4 follicles but not in F1 follicles. Neither BMP-6 alone, nor FSH/LH alone, affected 'basal' OB mRNA abundance. However, co-treatment with gonadotrophin and BMP-6 greatly increased beta(B)-subunit expression, the response being lowest in F1 follicles and greatest in F3/4 follicles. Collectively, these results support the hypothesis that intra-ovarian OMPs of thecal origin have a paracrine role in modulating granulosa cell function in the chicken in a preovulatory stage-dependent manner.

In vitro fermentation and prebiotic potential of novel low molecular weight polysaccharides derived from agar and alginate seaweeds

Fermentation properties and prebiotic potential of novel low molecular weight polysaccharides (LMWPs) derived from agar and alginate bearing seaweeds was investigated. Ten LMWPs were supplemented to pH, temperature controlled anaerobic batch cultures inoculated with human feces from three donors, in triplicate. Microbiota changes were monitored using Fluorescent in-situ hybridization and short chain fatty acids, the fermentation end products were analysed using gas chromatography. Of the ten LMWPs tested, Gelidium seaweed CC2253 of molecular weight 64.64 KDa showed a significant increase in bifidobacterial populations from log(10) 8.06 at 0 h to log(10) 8.55 at 24 h (p = 0.018). For total bacterial populations, alginate powder CC2238 produced a significant increase from log(10) 9.01 at 0 h to log(10) 9.58 at 24 h (p = 0.032). No changes were observed in the other bacterial groups tested viz. Bacteroides, Lactobacilli/Enterococci, Eubacterium rectale/Clostridium coccoides and Clostridium histolyticum. The polysaccharides also showed significant increases in total SCFA production, particularly acetic and propionic acids, indicating that they were readily fermented. In conclusion, some LMWPs derived from agar and alginate bearing seaweeds were fermented by gut bacteria and exhibited potential to be used a novel source of prebiotics.

Effects of single- and multi-strain probiotics on biofilm formation and in vitro adhesion to bladder cells by urinary tract pathogens

Inhibition of biofilm seems to be a major mechanism of urinary tract pathogen exclusion, related to, and possibly dependent upon, the probiotic ability to reduce environmental pH. Exclusion via competition of binding sites is a possible in vivo mechanism for these probiotics. If an additive or synergistic effect exists between strains within a mixture, it does not manifest itself in a greater effect through these two inhibitory mechanisms.

Bile salt composition is secondary to bile salt concentration in determining hydrocortisone and progesterone solubility in intestinal mimetic fluids.

Simulated intestinal fluids (SIFs) used to assay the solubility of orally administered drugs are typically based on a single bile salt; sodium taurocholate (STC). The aim of this study was to develop mimetic intestinal fluids with a closer similarity to physiological fluids than those reported to date by developing a mixed bile salt (MBS) system (STC, sodium glycodeoxycholate, sodium deoxycholate; 60:39:1) with different concentrations of lecithin, the preponderant intestinal phospholipid. Hydrocortisone and progesterone were used as model drugs to evaluate systematically the influence of SIF composition on solubility. Increasing total bile salt concentration from 0 to 30 mM increased hydrocortisone and progesterone solubility by 2- and ∼25-fold, respectively. Accordingly, higher solubilities were measured in the fed-state compared to the fasted-state SIFs. Progesterone showed the greatest increases in solubility in STC and MBS systems (2-7-fold) compared to hydrocortisone (no significant change; P>0.05) as lecithin concentration was increased. Overall, MBS systems gave similar solubility profiles to STC. In conclusion, the addenda of MBS and lecithin were found to be secondary to the influence of BS concentration. These data provide a foundation for the design of more bio-similar media for pivotal decision-guiding assays in drug development and quality control settings.

Annual pattern of plasma melatonin and progesterone concentrations in hair and wool ewe lambs kept under natural photoperiod at lower latitudes in the southern hemisphere

To study the annual pattern of plasma melatonin and progesterone concentrations in hair [Santa Ines (SI)] and wool [Romney Marsh (RM) and Suffolk (SU)] ewe lambs kept under natural photoperiods at 21 degrees 59'S, 12 ewe lambs (four/breed) were used. For melatonin, blood samples were collected monthly throughout the year at the onset (17:00, 19:00 and 21:00 hr) and end (04:00, 06:00 and 08:00 hr) of the night, and for progesterone the samples were collected in the morning, two to three times a week throughout the year. Plasma melatonin concentrations at different times of the day changed according to the season. In diurnal periods (17:00 and 8:00 hr) no seasonal differences were observed but they became evident in the nocturnal intervals (21:00 and 4:00 hr) and transitional night-day (6:00 hr) times. The patterns of melatonin secretion were higher in winter and autumn than in spring and summer. The patterns of plasma progesterone secretion were affected by interaction between breed and season. There was no seasonal variation in plasma progesterone concentrations for SI females. The progesterone pattern for RM and SU females varied with season. The plasma levels were higher in autumn and winter than in spring and summer. At 21 degrees 59'S hair and wool ewe lambs showed the same annual pattern of plasma melatonin concentration while the annual progesterone profiles were quite different. For SI females this pattern was constant along all seasons and for RM and SU females this pattern was higher during autumn and winter than spring and summer.

Effect of timing of estradiol benzoate administration upon synchronization of ovulation in suckling Nelore cows (Bos indicus) treated with a progesterone-releasing intravaginal device

The present study investigated how the timing of the administration of estradiol benzoate (EB) impacted the synchronization of ovulation in fixed-time artificial insemination protocols of cattle. To accomplish this, two experiments were conducted, with EB injection occurring at different times: at withdrawal of the progesterone-releasing (N) intravaginal device or 24 h later. The effectiveness of these times was compared by examining ovarian follicular dynamics (Experiment 1, n = 30) and conception rates (Experiment 2, n = 504). In Experiment 1, follicular dynamics was performed in 30 Nelore cows (Bos indicus) allocated into two groups. on a random day of the estrous cycle (Day 0), both groups received 2 mg of EB i.m. and a P4-releasing intravaginal device, which was removed on Day 8, when 400 IU of eCG and 150 mu g of PGF were administered. The control group (G-EB9; n = 15) received 1 mg of EB on Day 9, while Group EB8 (G-EB8; n = 15) received the same dose a day earlier. Ovarian ultrasonographic evaluations were performed every 8 h after device removal until ovulation. The timing of EB administration (Day 8 compared with Day 9) did affect the interval between P4 device removal to ovulation (59.4 +/- 2.0 h compared with 69.3 +/- 1.7 h) and maximum diameter of dominant (1.54 +/- 0.06 a cm compared with 1.71 +/- 0.05 b cm, P = 0.03) and ovulatory (1.46 +/- 0.05 a cm compared with 1.58 +/- 0.04 b cm, P < 0.01) follicles. In Experiment 2,504 suckling cows received the same treatment described in Experiment 1, but insemination was performed as follows: Group EB8-AI48h (G-EB8-AI48h; n = 119) and Group EB8-AI54h (G-EB8-AI54h; n = 134) received 1 mg of EB on Day 8 and FrAI was performed, respectively, 48 or 54 h after P4 device removal. Group EB9-AI48h (G-EB9-AI48h; n = 126) and Group EB9-AI54h (G-EB9-AI54h n = 125) received the same treatments and underwent the same FTAI protocols as G-EB8-AI48h and G-EB8-AI54h, respectively; however, EB was administered on Day 9. Conception rates were greater (P < 0.05) in G-EB9-AI54h 163.2% (79/125) a], G-EB9-AI48h [58.7% (74/126) a] and G-EB8-AI48h [58.8% (70/119) a] than in G-EB8-AI54h [34.3% (46/134) b]. We concluded that when EB administration occurred at device withdrawal (D8), the interval to ovulation shortened and dominant and ovulatory follicle diameters decreased. Furthermore, when EB treatment was performed 24 h after device removal, FTAI conducted at either 48 or 54 h resulted in similar conception rates. However, EB treatment on the same day as device withdrawal resulted in a lesser conception rate when FTAI was conducted 54 h after device removal. (C) 2007 Elsevier B.V. All rights reserved.

Evaluation of thyroxine (T4) and progesterone-17-alpha-OH circadian rhythm in swine females (Sus scrofa domestica - Linnaeus, 1758)

No presente trabalho foram utilizadas quatro fêmeas suínas, adultas, mestiças, não-gestantes e sem sinais clínicos de estro, criadas e mantidas sob condições industriais de criação. Objetivou-se avaliar a ocorrência de ritmicidade biológica circadiana para tiroxina e 17-alfa -OH progesterona. Os ensaios para dosagens hormonais foram executados utilizando-se a técnica de radioimunoensaio (RIE) em fase sólida e para isso foi empregado conjunto de reagentes comerciais (COAT-A-COUNT R). As análises séricas de tiroxina mostraram valores mais elevados ao redor das 15 horas, decrescendo a partir dai até atingir níveis menores no intervalo da zero às 4 horas. Quanto a 17-alfa -OH progesterona, observaram-se níveis mais elevados por volta das 3 horas, decrescendo gradativamente ao longo do dia, até atingir menor concentração no intervalo das 12 às 15 horas.

Fertility Rates Following Fixed-Time Artificial Insemination in Dairy Heifers in a Practical Progesterone-Based Protocol

Background: In bovines, more efficient management practices are important for maximizing profitability. In order to increase the pregnancy rates in artificial insemination (AI) programs, several hormonal protocols were developed to synchronize the follicular wave and the moment of ovulation in beef and dairy cattle. In dairy cattle, detection of estrus can be difficult due to a number of factors including the incidence of silent estrus. Hormonal treatments designed to control both luteal and follicular function has permitting efficient synchronizations of time of ovulation. Thus, the AI can be performed in a large number of animals on a fixed schedule without the need for detection of estrus. Using these management techniques, the fixed-time artificial insemination (TAI) can overcome the problem of accurate estrus detection and help in reducing the incidence of repeat breeding. In addition, with TAI in cattle operations, it is possible to facilitate management practices and commercialization, and to reduce the time and semen wasting with animals inseminated at incorrect times. The investigation of practical and efficient TAI protocols is important for reducing the labor and animal handling of TAI in dairy cattle, as well as for increasing the profitability of the cattle management system. This study was carried out in order to investigate the effectiveness of TAI in dairy heifers treated with a practical progesterone-based protocol.Materials, Methods & Results: This experiment was conducted at the university farm located in southwestern Brazil, during May 2009. Thirty-nine cycling crossbred dairy heifers were employed in this study. All animals received a single intramuscular injection of estradiol benzoate and intravaginal progesterone releasing device in a random stage of the estrous cycle (Day 0). on day 7 the animals were treated with PGF2a analogue and on day 9 the device was removed. Forty-eight hours after the device removal (day 11) a synthetic analogue of GnRH was administered and the animals were fixed-time artificially inseminated at the time of GnRH injection. The inseminations were performed using four different batches from the same Holstein bull. Among the heifers that were synchronized (87.2%), 30.8% ovulated until 24 h after TAI and 56.4% ovulated between 24 and 32 h after TAI. The conception rate was 61.5%. No effects of ovulation time in conception rates were detected. The conception rate from heifers that ovulated until 24 h after TAI was 58.3% and from heifers that ovulated between 24 and 32 h after TAI was 77.3%. The mean of ovulatory follicle in heifers that ovulated until 24 h was 14.3 mm and in heifers that ovulated between 24 and 32 h was 11.9 mm.Discussion: Taking together, the findings of the present study, along with those of others, emphasize the concept that development of practical methods for TAI offers significant advantages to dairy producers if conception rates are close or greater to those obtained after breeding at detected estrus. Thus, the results of the present study reinforce the possibility of making dairy cattle production more cost-effective using TAI. In conclusion, with the progesterone-based TAI protocol of the present experiment all synchronized animals ovulated up to 32 h after GnRH+TAI and no effects of ovulation time related to conception rate was detected. The exogenous control of luteal and follicular development facilitated the reproductive management and animal handling. Also, inseminating the heifers at the moment of GnRH injection in a progesterone-based TAI protocol is a practical strategy and provided satisfactory results regarding ovulation and conception rates in dairy heifers.

Monitoring ovarian cycles and pregnancy in brown brocket deer (Mazama gouazoubira) by measurement of fecal progesterone metabolites

In the present study, pregnancy and the estrous cycle were monitored in captive brown brocket deer (Mazama gouazoubira) by measuring fecal progestagens with a commercial enzyme immunoassay (EIA), along with behavioral data. Fecal samples were collected twice a week during pregnancy and daily during the estrous cycle and post-partum period. It was possible to distinguish between inter-luteal and luteal phases of the estrous cycle. Behavioral estrus corresponded with low concentrations of fecal progestagens. Samples from two consecutive cycles were available from five hinds, and the mean estrous cycle (n = 10) was 26.9 +/- 1.7 d (mean +/- S.E.M.). However, when two extreme cycles (34 and 37 d) were deleted, the mean estrous cycle was 24.7 +/- 1.2 d. Three animals became pregnant (gestation ranged from 208 to 215 d). After fertile breeding, progestagen concentration in these hinds remained among luteal phase concentrations throughout pregnancy, with the exception of a few peaks. Within 4 d post-partum, two hinds reached interluteal phase values, while one hind maintained luteal concentrations for at least 1 week. (C) 2005 Elsevier B.V. All rights reserved.

Evaluation of estrogen receptor alpha and beta and progesterone receptor expression and correlation with clinicopathologic factors and proliferative marker Ki-67 in breast cancers

To elucidate the molecular profile of hormonal steroid receptor status, we analyzed ER-alpha, ER-beta, and PGR mRNA and protein expression in 80 breast carcinomas using reverse transcriptase polymerase chain reaction (RT-PCR), quantitative RT-PCR, and immunohistochemical analysis. Qualitative analysis revealed positive expression of ER-alpha, ER-beta, and PGR mRNA in 48%, 59%, and 48% of the breast carcinomas, respectively. ER-alpha, ER-beta, and PGR transcript overexpression was observed in 51%, 0%, and 12% of the cases, respectively, whereas moderate or strong protein expression was detected in 68%, 78%, and 49% of the cases, respectively. Tumor grade was negatively correlated with transcript and protein levels of ER-alpha (P = .0169 and P = .0006, respectively) and PGR (P = .0034 and P = .0005, respectively). Similarly, proliferative index Ki-67 was negatively associated with transcript and protein levels of ER-alpha (P = .0006 and P < .0001, respectively) and PGR (P = .0258 and P =. 0005, respectively). These findings suggest that ER-alpha and PGR expression are associated with well-differentiated breast tumors and less directly related to cell proliferation. A significant statistical difference was observed between lymph node status and ER-beta protein expression (P = .0208). In ER-alpha-negative tumors, we detected a correlation between ER-beta protein expression and high levels of Ki-67. These data suggest that ER-beta could be a prognostic marker in human breast cancer. (C) 2008 Elsevier B.V. All rights reserved.