Reproductive cycle, ovarian development, and vertebrate-type steroids profile in the freshwater prawn Macrobrachium rosenbergii

This study is intended to characterize the ovarian development and hemolymph vertebrate-type steroids concentration during the reproductive cycle of the freshwater prawn, Macrobrachium rosenbergii. A five-stage classification based on the external observation of the ovary's size and color as seen through the tegument was used. The results showed the existence of a direct correspondence between the ovarian stages and the gonadosomatic index, the oöcyte diameter, and the characteristics of ovarian histology. In each stage total bleeding of the prawns was conducted and the hemolymph concentrations of 17β-estradiol (E 2), testosterone (T) and 17α-hydroxyprogesterone (17-OHP; conjugated and unconjugated) were determined by solid-phase radioimmunoassay (RIA). High levels of unconjugated 17-OHP, relatively constant concentrations of unconjugated T, and null concentration of unconjugated E 2 were found throughout the five stages considered. Nonetheless, low levels of E 2 were determined in all stages, in conjugated (glucoronide) form. The highest levels were obtained in stage II and III (18.14 ± 14.52 pg/ml hemolymph) and progressively declined in the other stages (7.53 ± 6.76 pg/ml hemolymph). These results point out the possible involvement of vertebrate-type steroids in the endocrine regulation of the Macrobrachium rosenbergii's ovarian cycle. To our knowledge this is the first study concerning the vertebrate-like steroid levels throughout the ovarian cycle and is a step needed to characterize the hemolymph profile of these steroids in order to elucidate the possible role in the regulation of the reproductive crustacean cycle.

Effects of atrazine on female Wistar rats: Morphological alterations in ovarian follicles and immunocytochemical labeling of 90 kDa heat shock protein

Fertility in female mammals may be affected by a variety of endocrine disrupters present in the environment. Herbicide atrazine is an example of endocrine disrupter employed in agriculture, which disrupts estrous cyclicity in rats. Aiming to characterize morphologically the effect of low and sublethal doses of atrazine on the ovaries of Wistar rats, in an effort to determine the possible intrafollicular target site through which this herbicide acts adult females were submitted to both subacute and subchronic treatments. Additionally, immunocytochemical labeling of 90 kDa heat shock protein (HSP90) was performed in order to evaluate the role played by this protein in the ovary, under stressed conditions induced by herbicide exposure. The results indicated that atrazine induced impaired folliculogenesis, increased follicular atresia and HSP90 depletion in female rats submitted to subacute treatment, while the subchronic treatment with low dose of atrazine could compromise the reproductive capacity reflected by the presence of multioocytic follicle and stress-inducible HSP90. © 2007 Elsevier Ltd. All rights reserved.

Scanning electron microscopic study of the in situ effect of salivary stimulation on erosion and abrasion in human and bovine enamel

This in situ study investigated, using scanning electron microscopy, the effect of stimulated saliva on the enamel surface of bovine and human substrates submitted to erosion followed by brushing abrasion immediately or after one hour. During 2 experimental 7-day crossover phases, 9 previously selected volunteers wore intraoral palatal devices, with 12 enamel specimens (6 human and 6 bovine). In the first phase, the volunteers immersed the device for 5 minutes in 150 ml of a cola drink, 4 times a day (8h00, 12h00, 16h00 and 20h00). Immediately after the immersions, no treatment was performed in 4 specimens (ERO), 4 other specimens were immediately brushed (0 min) using a fluoride dentifrice and the device was replaced into the mouth. After 60 min, the other 4 specimens were brushed. In the second phase, the procedures were repeated but, after the immersions, the volunteers stimulated the salivary flow rate by chewing a sugar-free gum for 30 min. Enamel superficial alterations of all specimens were then evaluated using a scanning electron microscope. Enamel prism core dissolution was seen on the surfaces submitted to erosion, while on those submitted to erosion and to abrasion (both at 0 and 60 min) a more homogeneous enamel surface was observed, probably due to the removal of the altered superficial prism layer. For all the other variables - enamel substrate and salivary stimulation the microscopic pattern of the enamel specimens was similar.

LH surge in Nelore cows (Bos indicus), after induced estrus or after ovarian superestimulation

Considering that there is limited information about the preovulatory LH surge in Zebu cattle (Bos indicus), the purpose of the present work was to assess the LH surge in Nelore cows during the estrous cycle and after ovarian superestimulation of ovarian follicular development with FSH. This information is particularly important to improve superovulatory protocols associated with fixed-time artificial insemination. Nelore cows (n = 12) had their estrus synchronized with an intravaginal device containing progesterone (CIDR-B ®) associated with estradiol benzoate administration (EB, 2.5 mg, i.m., Day 0). Eight days later all animals were treated with PGF2α (Day 8) in the morning (8:00 h) and at night, when CIDR devices were removed (20:00 h). Starting 38 h after the first PGF2α injection, blood sampling and ovarian ultrasonography took place every 4 h, during 37 consecutive hours. Frequent handling may have resulted in a stress-induced suppression of LH secretion resulting in only 3 of 12 cows having ovulations at 46.7 ± 4.9 and 72.3 ± 3.8 h, respectively, after removal of CIDR-B. Thirty days later, the same animals received the described hormonal treatment associated with FSH (Folltropin ®, total dose = 200 mg) administered twice a day, during 4 consecutive days, starting on Day 5. Thirty-six hours after the first injection of PGF2α, to minimize stress, only seven blood samples were collected at 4 h interval each, and ultrasonography was performed every 12 h until ovulation. In 11 of 12 cows (92%) the LH surge and ovulation were observed 34.6 ± 1.6 and 59.5 ± 1.9 h, respectively, after removal of progesterone source. The maximum values for LH in those animals were 19.0 ± 2.6 ng/ml (mean ± S.E.M.). It is concluded that, in Nelore cows submitted to a ovarian superstimulation protocol, the LH surge occurs approximately 35 h after removal of intravaginal device containing progesterone, and approximately 12 h before the LH surge observed after an induced estrus without ovarian superstimulation. © 2008 Elsevier B.V. All rights reserved.

Utilization of neuromuscular electrical stimulation in the acute phase of ankle immobilization at 90°: Study in rats

Objective: To evaluate the skeletal muscle glycogen content and plasmatic concentration of interleukin -6 (IL-6), interleukin-4 (IL-4), interleukin-10 (IL-10) and tumor necrosis factor-alpha (TNF-α) in rats submitted to electrical stimulation sessions during the first three days of ankle immobilization at the position of 90°. Methods: Albinomale Wistar rats(3-4 months) were maintained in vivarium. conditions with food and water ad libitum, Submitted to 12 h photoperiodic cycles of light/dark, and distributed into 7 experimental groups (n = 6): control(C), immobilized 1 day(I1) immobilized 1 day and electrically stimulated(IE1) immobilized 2 days(12), immobilized 2 days and electrically stimulated(IE2), immobilized 3 days(13) and immobilized 3 days and electrically stimulated(IE3). Groups I utilized an acrylic resin orthesis model and groups electrically stimulated (IE) utilized the orthesis and a session of electrotherapy by a Dualpex 961 (biphasic quadratic pulse, 10 Hz, 0.4 ms, 5.0 mA, one 20 min session a day). After the experimental period, the rats were anesthetized with pentobarbital sodium(40 mg/kg) and a blood sample was colleted to evaluate the plasmatic concentration of interleukins by means of the radioimmunoassay method. The soleus and the white portion of the gastrocnemius muscle were colleted for glycogen reserves analysis(GLY). Other groups of rats were used to apply the glucose tolerance test(GTT) and insulin tolerance test(ITT). For statistical analysis, the Kolmogorov-Smirnov normality test followed by ANOVA and the Tukey tests were utilized, with a critical level established at 5%. Results: In ITT test, groups IE enhanced the skeletal muscle glucose uptake, but no changes were observed in GTT after the therapy session, which indicates that electrical stimulation is a sensibilizing method to augment skeletal muscle glucose uptake. The GLY reserves were reduced in I groups, which indicate that disuse altered insulin sensitivity and compromised energetic homeostasis. However. the IE groups displayed an augment in GLY content, suggesting that electrical stimulation restores the enzymatic pathways altered by immobilization. The improvement in GLY was accompanied by an elevation of the plasmatic concentration of IL-6 and TNF-α, showing the participation of these interleukins in the control of metabolic profile. Plasmatic concentrations of IL-10 were elevated only after 3 days of IE while IL-4 did not display any modifications. Conclusion: The results suggest that neuromuscular electricaf stimulation is an important toot in the maintenance of energetic, conditions of musculature submitted to immobilization, and presents multifactor mechanisms linked to interleukins action that converge to maintain the energetic equilibrium of the tissue in disuse.

Mare's folliculogenesis: Assessment of ovarian and perifollicular vascular perfusion by doppler ultrasound

Background: Ultrasound Doppler is a new technology that has recently been used in large animal reproduction. As the conventional ultrasound (B-mode) it is a noninvasive technique, but with the advantage of allows the assessment of the hemodynamic of reproductive tract in real time. The observation of important features of the vascularization and changes that occur during physiological processes that were not previously seen on B-mode encourage a reassessment of the concepts already established on the events of the reproductive physiology of animals and their applications. Review: In attempt to re-understand the equine reproductive physiology and finding practical uses to this new technique, authors showed that, during the follicular deviation, features are observed by Doppler before being observed under B-mode ultrasound like changes in the speed of the blood flow two days before deviation of the follicle size and one day before the increase in blood flow area of the follicular wall. According to another study ovulation is characterized by a decreased blood flow of the follicular wall in the last four hours preceding it, as well as the serration of the granulosa layer and formation of a non vascularized apex, but in our ongoing study, the ability to decrease the vascularity was not found. Very vascularized follicles are associated with higher rates of oocyte maturation and pregnant that does less vascularized follicles in the preovulatory phase. Those follicles that have septated evacuation (or prolonged) showed more vascularization and serration of the granulosa one hour before ovulation than follicles that ovulate normally, and this vascularization includes the apex of the follicle, the follicular wall portion that is not vascularized in normal ovulation. Another study reported that hemorrhagic follicles have better vascularization of the follicular wall on the days preceding ovulation than follicles destined to ovulate. Some authors also showed that anovulatory follicles grow in size at the same rate as ovulatory follicles, but the percentage of vascularization of its wall is much smaller at 35 mm. Another study characterized that the vascular wall of the follicle that results in the first ovulation of the year is much smaller on the day before ovulation than the number of vessels present in a follicle that will ovulate in the middle of the breeding season. In these cases, the use of Doppler ultrasound can help to prevent economic losses as insemination of mares in cycles that are not able of resulting in pregnancy. This review aims to gather the information found in the literature about the characteristics of follicular hemodynamic of mares taking into account moments of deviation in follicle size, ovulation, ovulation failure and follicular viability. Conclusion: The Doppler technology has the potential to provide important information about the follicular environment and thus be used in practice in search of the perfect equine reproductive management, achieving better utilization of genetic material and increasing the financial return. The use of this new tool opens a large area for several interesting studies that will contribute to the knowledge of the physiological events of the mare for that this technique can soon be effectively applied.

Porphyromonas gingivalis LPS stimulation downregulates DNMT1, DNMT3a, and JMJD3 gene expression levels in human HaCaT keratinocytes

Objective: The role of epigenetic regulation in inflammatory diseases such as periodontitis is poorly known. The aim of this study was to assess whether Porphyromonas gingivalis lipopolysaccharide (LPS) can modulate gene expression levels of the some enzymes that promote epigenetic events in cultures of the human keratinocytes and gingival fibroblasts. In addition, the same enzymes were evaluated in gingival samples from healthy and periodontitis-affected individuals. Materials and methods: Primary gingival fibroblast and keratinocyte (HaCaT) cultures were treated with medium containing P. gingivalis LPS or P. gingivalis LPS vehicle for 24 h. After this period, cell viability was assessed by MTT test and total RNA extracted to evaluate gene expression levels of the following enzymes by qRT-PCR: DNA methyltransferase 1 (DNMT1), DNA methyltransferase 3a (DNMT3a), histone demethylases Jumonji domain containing 3 (JMJD3) and ubiquitously transcribed tetratricopeptide repeat, X chromosome (UTX). To evaluate gene expression in healthy and periodontitis-affected individuals, total RNA was extracted from biopsies of gingival tissue from healthy and periodontitis sites, and gene expression of DNMT1, DNAMT3a, JMJD3, and UTX was evaluated by qRT-PCR. Results: No significant differences were found in the gene expression analysis between healthy and periodontitis-affected gingival samples. The results showed that LPS downregulated DNMT1 (p < 0. 05), DNMT3a (p < 0. 05), and JMJD3 (p < 0. 01) gene expression in HaCaT cells, but no modulation was observed in gingival fibroblasts. Conclusion: P. gingivalis LPS exposure to human HaCaT keratinocytes downregulates gene expression of the enzymes that promote epigenetic events. Clinical relevance: The advance knowledge about epigenetic modifications caused by periodontopathogens may to possibly led to the development of new periodontal therapies. © 2012 Springer-Verlag.

Ovarian response to porcine FSH in association with ablation-induced or spontaneous follicular wave development during the estrous cycle in crossbred and Brazilian Warmblood mares

The primary objective of this study was to examine the follicular and ovulatory responses following treatment with pFSH in association with ablation-induced or spontaneous follicular wave emergence or follicle deviation during diestrus in crossbred (Mangalarga × Arabian) and Brazilian Warmblood mares with a propensity for spontaneous multiple ovulations; secondary considerations were given to the collection of embryos In Experiment 1, crossbred mares were administered (im) saline (control, n= 7) or pFSH (25 mg) when the largest follicle of the ablation-induced follicular wave reached ≥13 mm (n= 7) or ≥20 mm (n= 7) or, after pre-treatment ovulation (Day 0) on Day 6 (n= 7) In Experiment 2, crossbred mares were administered (im) saline (control, n= 10) or a larger dose of pFSH (50 mg, n= 7) when the largest follicle of the ablation-induced follicular wave reached ≥13 mm In Experiment 3, Brazilian Warmblood mares were administered (im) saline (control, n= 7), pFSH (25 mg, n= 7 or 50 mg, n= 5) or EPE (12.5 mg, n= 7) as a positive control on Day 6 Ultrasonic technology was used to ablate all follicles ≥8 mm and to monitor follicular development and detect ovulation Treatment with pFSH or EPE was done twice daily until the largest follicle reached ≥32 mm; thereafter, hCG (2500 IU) was administered (iv) when the largest follicle reached ≥35 mm Artificial insemination was done 12 h after hCG and embryo collections were done 8 d after post-treatment ovulations In Experiments 1 and 2, treatment of crossbred mares with pFSH post-ablation in association with the expected time of wave emergence or follicle deviation did not (P> 0.05) enhance the follicular or ovulatory responses or collection of embryos compared to controls In Experiment 3, although the enhanced ovulatory response of mares to EPE at the expected time of spontaneous wave emergence was not different (P> 0.05) from controls, it was greater (P< 0.05) than the response to pFSH In conclusion, the novelty of using follicle ablation prior to pFSH treatment at the time of wave emergence or follicle deviation did not enhance the follicular or ovulatory responses or collection of embryos to treatment in crossbred mares In addition, the hypothesis that Brazilian Warmblood mares with a greater propensity for spontaneous multiple ovulations are as responsive to pFSH compared to EPE was not supported Thus, the combined experimental results of the present study continue to support the general consensus that pFSH is relatively ineffective for follicular superstimulation/superovulation in mares © 2012 Elsevier B.V.

Ovarian Tumor in a Mare-Thecoma-Case Report

Thecomas are tumors originating from the ovarian stroma and of rare occurrence in the equine species. Clinical, ultrasonographic, endocrine, and histopathologic descriptions of thecoma are scarce. This article aims to describe a case of ovarian changes in a mare with anomalous growth of the right ovary and anestrus for 1 year. After surgical removal, the ovarian growth was diagnosed as a thecoma. Histopathological findings were consistent with the findings previously described in the literature, characteristic of a thecoma, and decisive for the correct diagnosis. © 2013 Elsevier Inc.

Ovarian Fibrothecoma in Mare - Case Report

A 5-year-old Mangalarga mare was referred to the Veterinary Teaching Hospital of the Veterinary Medical School in Botucatu with a complaint of a persistent 2-month ovarian follicle which had grown progressively to 20 cm in diameter. Ultrasonography showed a circular, cystic structure in the right ovary, whose wall was thickened and interior was filled with anechoic content. Ovarian neoplasia was suspected, and a unilateral ovariectomy was performed through the paralumbar fossa, using general anesthesia through inhalation. Histopathological evaluation concluded that the ovarian enlargement was caused by a fibrothecoma. The present report describes features of fibrothecoma, which is extremely rare in the equine species. © 2013 Elsevier Inc. All rights reserved.

Hormonal treatment of dairy goats affected by hydrometra associated or not with ovarian follicular cyst

Hydrometra is considered a very important pathological condition, because it represents one of the main causes of temporary infertility in dairy goats. The objective was (i) to evaluate a protocol for the treatment of hydrometra associated (n=2) or not (n=17) with follicular ovarian cyst in 19 dairy goats and (ii) to assess its reproductive efficiency after treatment. For this purpose, 10. mg of dinoprost (PGF) divided in two equal doses were administered to all animals intravulvosubmucosally on Days 0 and 10. In addition, 500. IU hCG were administered on Day 7. Ultrasound exams were performed in all females from Days 0 to 3, 7 and 10 to 13 of treatment, in order to evaluate uterus drainage after each treatment. Goats were monitored for estrus after both treatments and mated after the second dose of PGF. Blood samples were collected from 11 goats to determine plasma progesterone concentrations before, during and after treatment. Of the 19 goats treated, 16 lost weight after the first dose, probably due to uterine discharge. Complete drainage of uterine fluid was observed in 11/19 (57.9%) and 17/19 (89.5%) after the first and second doses, respectively. Afterwards, we diagnosed 2 more goats with follicular cysts, for a total of 21.1% (4/19) of animals exhibiting hydrometra and ovarian cyst concomitantly. In one doe the diagnosis was on Day 2 and in the other on Day 11. All does showed progesterone concentrations superior to 1. ng/mL at Day 0, with an average of 10.6 ± 1.4. ng/mL. Out of the 10 goats mated, only two became pregnant after treatment, corresponding to 10.5% of the total (2/19). Although prostaglandin was effective to drain the uterine fluid and led to the onset of estrus, it did not improve the pregnancy rate. The use of hCG in female goats was not effective in luteinizing the cysts. It can be concluded that hydrometra alone or associated with ovarian follicular cyst may adversely affect goat reproductive performance. © 2012 Elsevier B.V.

The ovarian cycle histochemistry and its relationship with hepatopancreas weight in the blue crab Callinectes danae (Crustacea: Portunidae)

Zara, F.J., Gaeta, H.H., Costa, T.M., Toyama, M.H. and Caetano, F.H. 2011. The ovarian cycle histochemistry and its relationship with hepatopancreas weight in the blue crab Callinectes danae (Crustacea: Portunidae). -Acta Zoologica (Stockholm) 00:1-13. Several studies use macroscopic patterns of the ovarian development in crustaceans. Here, we examined the relationship between ovary histochemistry, changes in gonadosomatic and hepatosomatic indices against the macroscopic pattern of the ovarian development in Callinectes danae. Animals were collected in the south coast of São Paulo State, Brazil. Ovaries were macroscopically classified as juvenile, rudimentary, developing, intermediary, mature, and rudimentary ovigerous. Samples were fixed in 4% paraformaldehyde, processed for historesin, and stained with HE, protein, and neutral and acid polysaccharides detection. The juvenile oocytes are not enclosed by follicular cells and have fewer yolk nuclei being less intense in PAS reactivity than rudimentary oocytes. Developing oocytes show yolk granules and thick follicular cells. Yolk granules were positive for proteins and neutral polysaccharides. The intermediary stage is marked by a qualitative increase in yolk granules and the onset of chorion formation. In mature oocytes, cytoplasm is completely filled by yolk granules and the chorion is completely formed. Ovigerous ovaries have several atretic follicles and large quantities of hemocytes in the process of tissue reorganization. In C. danae, the changes in cell, goandosomatic and hepatosomatic indices coinciding with macroscopic observations and any combination of different macroscopic stages in a single pattern should be avoided. © 2011 The Royal Swedish Academy of Sciences.

Ovarian morphology and oogenesis of Leptodesmus dentellus (Diplopoda, Polydesmida, Chelodesmidae)

This study presents the ovarian morphology and the dynamics of the vitellogenesis process in the oocytes of the diplopod Leptodesmus dentellus. The oocytes are arranged in clusters called ovisacs which are distributed in pairs along the midline of the body forming the ovary. Regions similar to the germarium appear paired in the anterior region of the ovary; however, the development of the oocytes of this species does not follow a regionalisation in the reproductive organ. Cells in three developmental stages are found throughout the length of the ovary. Calcium, proteins, lipids and neutral polysaccharides were detected in the oocytes of L. dentellus. The polysaccharides and the proteins found in the oocytes have a double origin: endogenous, with the participation of the germinative vesicle, and exogenous, from follicular epithelium. The origin of the lipids is exogenous, i.e., they are incorporated into the oocytes, probably derived from the perivisceral fat body, which are highly developed in this region. The deposition of calcium is pre-vitellogenic and probably functions as a reserve during the juvenile stages. © 2012 Koninklijke Brill NV, Leiden.

The effects of different styles of musical auditory stimulation on cardiac autonomic regulation in healthy women

The literature investigated the effects of chronic baroque music auditory stimulation on the cardiovascular system. However, it lacks in the literature the acute effects of different styles of music on cardiac autonomic regulation. To evaluate the acute effects of baroque and heavy metal music on heart rate variability (HRV) in women. The study was performed in 21 healthy women between 18 and 30 years old. We excluded persons with previous experience with music instrument and those who had affinity with the song styles. All procedures were performed in the same sound-proof room. We analyzed HRV in the time (standard deviation of normal-to-normal respiratory rate (RR) intervals, root-mean square of differences between adjacent normal RR intervals in a time interval, and the percentage of adjacent RR intervals with a difference of duration greater than 50 ms) and frequency (low frequency [LF], high frequency [HF], and LF/HF ratio) domains. HRV was recorded at rest for 10 min. Subsequently they were exposed to baroque or heavy metal music for 5 min through an earphone. After the first music exposure they remained at rest for more 5 min and them they were exposed again to baroque or heavy metal music. The sequence of songs was randomized for each individual. The power analysis provided a minimal number of 18 subjects. Shapiro-Wilk to verify normality of data and analysis of variance for repeated measures followed by the Bonferroni test for parametric variables and Friedman's followed by the Dunn's post-test for non-parametric distributions. During the analysis of the time-domain indices were not changed. In the frequency-domain analysis, the LF in absolute units was reduced during the heavy metal music stimulation compared to control. Acute exposure to heavy metal music affected the sympathetic activity in healthy women.