Vaginal Plug Research Articles

Female Reproductive Performance in the Mouse: Effect of Oral Melatonin.

Although melatonin has some of the broadest ranges of actions on the physiology of vertebrates, especially on their reproductive processes, the mechanism by which melatonin regulates animal reproduction is still incompletely understood. This study was designed to determine the effect of oral melatonin on the reproductive performance of female mice. Female ICR mice (7 weeks old) were given melatonin-containing water (3, 30 and 300 μg/mL; melatonin) or water only (control) until 10 weeks of age. Then, some of the mice were successfully mated (confirmed by vaginal plugs), and the number of live births and their weights were recorded. Some mice were used for a histological analysis of the number of follicles in the ovaries. Others were used for oocyte collection after superovulation, and in vitro fertilization (IVF) was performed. The mRNA expression of the apopotosis-related genes (BAX, BCL2) in the IVF embryos were analyzed. After melatonin administration, the mice showed similar serum melatonin levels to that of the control. The number of antral follicles per mm2 unit area in the 30 μg/mL melatonin-treated group (14.60) was significantly higher than that of the control (7.78), which was lower than that of the 3 μg/mL melatonin-treated group (12.29). The litter size was significantly higher in the 3 μg/mL melatonin-treated group (15.5) than in the control (14.3). After IVF, the hatched blastocyst formation rate in the 30 μg/mL melatonin-treated group (85.70%) was significantly higher than that of the control (72.10%), and it was the same for the BCL2/BAX expression ratio. Although oral melatonin did not appear to have an effect on the serum melatonin rhythm in the mouse, melatonin did increase litter size at the 3 μg/mL dose level, and improved the developmental competency of IVF embryos at the 30 μg/mL level.

Deposition, removal and production site of the amorphous mating plug in the spider Philodromus cespitum

In order to avoid sperm competition, males of many taxa apply physical barriers, so-called mating plugs, into female genitalia. Females may control which males deposit a plug through pre-copulatory mate choice or by influencing plug efficacy to avoid costs imposed by plugging. However, subsequent suitors might remove the plugs. We investigated behavioural and morphological aspects of plug deposition and removal in a promiscuous spider, Philodromus cespitum (Philodromidae). We performed mating trials to investigate factors affecting plugging. To identify the plug origin, we conducted a morphological analysis using 3D X-ray microtomography and histology of the male copulatory organ and the female genital tract. In P. cespitum, the plug material is produced in the male genital bulb and transferred to the female together with sperm. The copulation is brief and terminated by the female. After mating, plugging material was found in the genital atrium of all females, covering it to a varying degree (10-100%). The extent of coverage was associated with the duration of movements of male copulatory organ connected with sperm transfer (i.e. full haematodochal expansions) and with the number of taps a male delivered with his legs to the female during courtship. Males larger than the female performed more tapping movements. Mating trials with plugged females revealed that males could remove plugs partly or entirely. Removal success increased with increasing foreleg length ratio between the male who removed the plug and the one who deposited it. We discuss our results in the light of the potential female control of plug deposition and removal.

Tokishakuyakusan, a traditional Japanese medicine (Kampo) mitigates iNKT cell-mediated pregnancy loss in mice.

Tokishakuyakusan (TSS) is a traditional herbal medicine that has been used empirically to prevent recurrent pregnancy loss. Its mode of action remains unclear. With their potent capacity to produce cytokines, invariant natural killer (iNKT) cells are involved in the control of fetomaternal immunity in early gestation. This study aimed to clarify the effect of TSS on iNKT cell activities in a well-studied murine miscarriage model. Pregnant mice were fed 1% TSS-containing or control diet from the day of vaginal plug formation. Alpha-galactosylceramide (AGC) was administered intraperitoneally to the pregnant mice at day 9.5 postcoitus (pc) to stimulate iNKT cells. Peripheral cytokine levels were evaluated using cytokine arrays. The percentage of iNKT cells among splenocytes was examined by flow cytometric analysis. The incidence of pregnancy loss was assessed at day 12.5 pc. The ratio of fetal resorptions to total conceptuses was significantly higher in the group exposed to TSS (34%) than in controls (78%). A rapid and robust surge in inflammatory cytokines, including IFN-γ and TNF-α, was detected in the peripheral blood of control animals 2hours after AGC administration. This peripheral cytokine induction was significantly attenuated in the TSS-fed group compared with the control. The percentage of iNKT cells among total splenocytes was lower in the TSS-fed group than in controls. The findings in this study suggest that the inhibitory effects of TSS on pregnancy loss may involve immune modulation of iNKT cells during early pregnancy.

An investigation into the male reproductive system of two freshwater crabs from the Amazon: is there a sperm plug or packet formation?

ABSTRACTThe male reproductive system of Amazon freshwater crabs Sylviocarcinus pictus and Dilocarcinus pagei were described and compared in terms of anatomy and histochemistry In addition, sperm transfer experiments of D. pagei with sequential copulations were performed and the anatomy of the seminal receptacle was checked. The anatomy of the male reproductive system of both species is an inverted ‘U’ shape, connected by the testes, from which the pair of vas deferens divided into anterior, median and posterior regions depart. Sylviocarcinus pictus and D. pagei revealed coenospermia, which differed from the families of freshwater crabs in Southeast Asia. The histochemical composition of the seminal fluid is similar in both Trichodactylidae, following the pattern described for most Brachyura. Through the copulation experiments and anatomical results, these animals do not show a sperm plug, but rather form different sperm packs when engaging in multiple copulations with different males.

Quantitative evolutionary proteomics of seminal fluid from primates with different mating systems

BackgroundGenomic data from various organisms have been used to study how sexual selection has shaped genetic diversity in reproductive proteins, and in particular, to elucidate how mating systems may have influenced evolution at the molecular and phenotypic levels. However, large-scale proteomic data including protein identifications and abundances are only now entering the field of evolutionary and comparative genomics. Variation in both protein sequence and expression level may play important roles in the evolution of sexual traits and behaviors.ResultsHere, we broadly analyze the components of seminal fluid from primates with diverse mating systems ranging from monogamous to polygynous, and include genomics, proteomics, phylogenetic and quantitative characters into our framework. Our analyses show that seminal fluid proteins are undergoing rapid evolution and some of these quickly evolving proteins may be influenced by sexual selection. Through evolutionary analyses and protein abundance differences, we identified 84 genes whose evolutionary rates or expression levels were correlated with mating system and other sexual characters. We found that many proteins differ in abundance between monogamous and polygynous primate mating systems. Many of these proteins are enriched in the copulatory plug pathway, which suggests that post-zygotic selective barriers are important regardless of mating system type.ConclusionsThis work is the first to comprehensively compare seminal fluid proteins between human and non-human primates using high-throughput proteomics. Our findings highlight the impact of mating system variation on seminal fluid protein evolution and abundance.

COMPARATIVE CHARACTERISTICS OF THERAPEUTIC EFFECTS OF CRYOEXTRACT PLACENTA AND MESENCHIMAL STEM CELLS OF FATTY TISSUE IN TREATMENT OF PREMATURE FAILURE OF OVARIANS CAUSED BY CHEMOTHERAPY IN EXPERIMENT

Premature ovarian failure (POF) is a common consequence for woman after treatment of oncological diseases. Nowadays the actual task is to find new methods to restore and preserve the ovarian function in women after chemotherapy (CT).The aim of the research was to compare the therapeutic effects of cryoextract of placenta (CP) and mesenchymal stem cells of adipose tissue (MSCs AT) in restoring the sexual function in POF after CT in the experiment.Materials and methods: POF was modeled on female BALB \ c mice. Animals were divided into 4 groups of 15 females with a weight of 20.1 ± 1.1 g with a regular estrous cycle: 1 – intact group, 2 - group with modeled POF, 3 - group with modeled POF and treated with CP (0.01 g in / m 1 p / day for 5 days). 4-group with modeled POF and treated with MSCs AT (100,000 cells intraperitoneally at once). We studied the weight, estrous cycle, sexual activity, morphological changes in the reproductive organs in animals.Results and discussion: After CT weight was reliably reduced in all mice, the estrous cycle became monophasic, there were no vaginal plugs. In the 2nd group of animals, without treatment, weight and activity were restored at 6 weeks after CT, the estrus cycle - after 5 weeks in 40% of animals, sexual activity in 6 weeks in 30% of mice. In 3 group of animals, weight and activity were restored after 3 weeks, estrous cycle in 70% animals after 4 weeks, sexual activity - in 5 weeks. In 4 group of animals treated with MSCs AT, weight and activity were restored after 3 weeks, the estrous cycle in 85% mice after 4 weeks, sexual activity - after 5 weeks. Histological examination indicated: in 2 group of animals ovaries were reduced in size and structural elements in them were absence. In uteri there were atrophic changes. In groups of animals treated with CP and MSCs AT, there were lack of typical elements in ovaries, separate follicle-like structures were observed, the structure of uteri was close to intact.Conclusions: In the experiment KP and MSCs AT efficacy when treating the POF, induced by CT.

The occurrence and function of the sphragis in the zerynthiine genera Zerynthia, Allancastria and Bhutanitis (Lepidoptera: Papilionoidea: Papilionidae)

ABSTRACTIn some butterfly species males attach a large external mating plug termed a sphragis to the female abdomen during mating. This is derived from male accessory secretions and covers the female ostium bursae and surrounding areas, thus preventing or delaying remating. Specimens of all 12 species of the genera Zerynthia, Allancastria and Bhutanitis (Lepidoptera: papilionidae), which form a natural clade within the Zerynthiini, were examined for presence or absence of a sphragis and their male and female genitalia were studied. In all three genera female genitalia lack a typical sinus vaginalis and the sterigma is modified to form an exposed, shiny, well-sclerotized genital plate, derived from the fusion and expansion of the lamellae ante- and postvaginales. The exposed ostium bursae is situated near the posterior end of the genital plate in Zerynthia, whereas in Allancastria and Bhutanitis it is near the anterior end. A crude irregularly formed sphragis was found at least facultatively in all species. The sphragides of Zerynthia and Bhutanitis were generally poorly developed, in most cases only partially covering the female genital plate. In Allancastria the sphragis mostly covered the genital plate entirely, and generally incorporated numerous long scales derived from the male’s 8th abdominal segment; scales were sometimes sparse or absent, probably due to depletion from repeated matings by males. In Zerynthia, males lacked the dense terminal abdominal tuft of elongated scales found in Allancastria, and their sphragis lacked scales. The sphragis of Bhutanitis thaidina incorporated scales from the male valves, whereas in the B. lidderdalii sphragis (and probably B. ludlowi) the scales derived from the male’s 8th abdominal segment. The role of the scales and possible reasons for the difference in the development of the sphragis among these genera are discussed.

Gestational diabetes influences the expression of hypertrophic genes in left ventricle of rat's offspring.

Objective(s):Gestational diabetes increases the risk of congenital heart disease in the offspring, but the molecular mechanism underlying this process remains unclear. Therefore, the current study was conducted to assess the effects of induced gestational diabetes on expression of some involved genes in cardiac hypertrophy in the offspring of diabetic rats.Materials and Methods:Diabetes was induced in 40 adult Wistar rats by intraperitoneal injection of 45 mg/kg of streptozotocin. The day of appearance of the vaginal plug was assumed as day zero of gestation for inducing diabetes. After pregnancy, the offspring was maintained until they reach the age of 12 weeks. Then, their hearts were excised and were sectioned for molecular study. We analyzed the expression pattern of some hypertrophic genes by the quantitative real-time RT-PCR.Results:The mRNA expression levels of all studied genes including c-jun, c-fos, c-myc, alpha-myosin heavy chain (α-MHC), atrial natriuretic factor (ANF) and β-MHC, which are important in cardiomyocyte hypertrophy, were higher in the offspring of the diabetic group compared to controls. Significant differences were found for β-MHC and c-myc with P<0.01 and for α-MHC and c-fos with P<0.05.Conclusion:Gestational diabetes upregulates expression of c-jun, c-fos c-myc, α-MHC, ANF and β-MHC genes that are involved in cardiac hypertrophy in the offspring of diabetic rats.

Induction of Zika specific CD8 T cell responses in the placenta after heat shock protein gp96-Ig vaccination

Abstract Heat shock protein (HSP) gp96 is a biological adjuvant that activates antigen-presenting cells and simultaneously delivers antigen specific peptides to MHC I for CD8 T cell activation. Here, we have transformed human embryonic kidney cell line (HEK-293) into powerful miniature osmotic pumps that continually secrete engineered form of gp96, gp96-Ig, along with its chaperoned antigens (ZIKV envelope immunogenic peptides) to induce ZIKV specific CD8+ cytotoxic T cell immune responses. B6 females were mated with B6 or BALB/c males and gestation day (GD) 0.5 was determined by the presence of vaginal plug. At GD 7.5 one million of irradiated 293-gp96-Ig secreting cells were injected subcutaneously (SC). Mice were followed up until the end of pregnancy (GD 19.5–21.5) or humanely euthanized 5 days after vaccination (GD 12.5). Frequency of CD8+ T cells was determent by flow cytometry, immunohistochemistry and antigen, ZIKV envelope specificity was confirmed by in vitro ZIKV peptide stimulation and by intracellular cytokine staining. We found that vaccinated mice have the same number of live-born pups as control mice without pathological changes in placenta or in fetus. This is the first finding that confirms the presence of gp96-Ig-ZIKV-induced CD8 T cells around arteries and larger blood vessels in the maternal part of the placenta, decidua. Furthermore, secreted gp96 vaccine induces very high frequency of ZIKV specific CD8 T cells in the maternal decidua (15%) without compromising placental or fetal viability. In summary, we have succeeded in generating vaccine that is expressing gp96-Ig and ZIKV envelope protein and also we confirmed that vaccination with secreted gp96-Ig-ZIKA is safe and induces CD8 T cell responses in maternal decidua.

Maternal exposure to Bisphenol‐A Induces Epigenetic Changes in metabolic syndrome markers of Offspring

Environmental factors exert significant effects of on disease progression later in the life, when exposed in‐utero. Bisphenol A (BPA) is a monomer of plastic products widely used in everyday life. BPA shows hormone mimicking properties, and its adverse effects on endocrine glands and hormones may affect developmental embryology of offspring. Bisphenol A may possibly have an epigenetic role in onset and progression of several adult diseases related to metabolic system. To determine the epigenetic effects of in‐utero Bisphenol A on markers related to metabolic syndrome in mice offspring. This study was conducted at department of Anatomy Liaquat University of Medical &amp; Health Sciences Jamshoro and Animal house of Sindh Agriculture University Tando Jam Pakistan after Ethical approval. Adult female BALB/c strains mice were divided in three groups each comprised of 10 female mice– G1 (controls), G2 (ethanol 0.10 ml i.p) and G3 (BPA 10 mg i.p). Animals were housed individually in polypropylene cages, in standard lighting, controlled temperature and humidity conditions. Control group was fed ad libitum. After mating, the positive vaginal plug was checked to confirm pregnancy. Offspring body weight, behavior, movement, blood glucose; serum insulin, cholesterol and HDLc were measured at 5 months age and at 15 months age before sacrifice of animals. Pancreas, kidney, adipose tissue and uterine tissue were stained with H &amp; E staining and Immunohistochemistry and examined under microscopy. Data was analyzed on SPSS 22.0 (at 95% confidence interval). Offspring of group G3 reveals abnormal changes in behavior and movement. Body weight, blood glucose, serum insulin, cholesterol and HDLc were high compared to controls (P=0.0001). Islets of Lanherghan's, Kidney and Uterine tissue reveals abnormal anatomical changes with anti‐ Islet‐1 Ig, Ksp cadherin, and anti‐ MLH antibody respectively. In utero exposure of endocrine disruptor Bisphenol A exerts Diabetogenic and Atherogenic effects with abnormal immunohistochemistry changes in endocrine pancreas, kidney and uterus.Support or Funding InformationThis study was funded by Liaquat University of Medical &amp; Health Sciences, Jamshoro, PakistanThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Prenatal Exposure to Bisphenol A Analogues on Male Reproductive Functions in Mice.

This study was performed to examine whether prenatal exposure to bisphenol (BP) A analogues, BPE and BPS, negatively impacts male reproductive functions and testis development using mice as a model. CD-1 mice were orally exposed to control treatment (corn oil), BPA, BPE, and BPS (0.5, 20, or 50 µg/kg/day) from gestational day 11 (the presence of vaginal plug = 1) to birth. Although sperm counts were significantly reduced by BPA, BPE, or BPS on postnatal day (PND) 60, only BPE or BPS exposure remained low for sperm motility. Exposure to BPA, BPE, and BPS disrupted the progression of germ cell development, as morphometric analyses exhibited an abnormal distribution of the stages of spermatogenesis. Furthermore, BPS with a dose of 50 µg/kg/day increased a level of serum estradiol-17β in adult males. On PND12, BPE and BPS significantly increased TUNEL positive cells in neonatal testis, following disruption of the expression of apoptosis, autophagy, and oxidative stress-related factors. In addition, the expression of methyltransferases for DNA methylation and histone modification was also affected by prenatal exposure to BPA, BPE, or BPS in neonatal testis. These results suggest that prenatal exposure to BPE and BPS with physiologically relevant doses affects male reproductive functions probably due to spermatogenic defect in the developing testis.

Influence of polygamous versus monogamous mating on embryo production in four different strains of mice after superovulatory treatment

We determined the effect of monogamous or polygamous mating with 2 females on vaginal plug (VP) rate, embryo donors (ED), 2-cell embryo production, and male performance after superovulation of females aging 24d or 45-48d. C57BL/6NCrl (B6N), BALB/cAnCrl (BALB/cN), FVB/NCrl (FVB/N), and Crl:CD1(ICR) (CD-1) females received 5 IU eCG and 5 IU hCG (24d) or 7.5 IU eCG and 7.5 IU hCG (45-48d) 48 h apart. After the hCG injection, females were paired with males, which alternated weekly in monogamous or polygamous mating. Significant differences in the percentage of VP-positive females between monogamous and polygamous mating were observed for B6N (71% vs. 49%), FVB/N (77% vs. 51%), and CD-1 (90% vs. 67%) at 45-48d. BALB/cN and CD-1 showed higher VP rates than B6N and FVB/N. A significantly higher percentage of ED was found for monogamous than for polygamous mating for FVB/N (87% vs. 61%) at 24d and for B6N (91% vs. 53%) and CD-1 (90% vs. 68%) at 45-48d. In all strains of mice and in both age groups, no significant differences were observed in the number of intact 2-cells per VP-positive female, ED or treated female between monogamous and polygamous mating except in the B6N strain where monogamous mating resulted in a significantly higher number of intact 2-cell embryos per treated female than polygamous mating at both ages. The present results imply that polygamous mating can be implemented for 2-cell embryo production in all strains studied except for B6N when all females are euthanized. However, when only VP+ females are sacrificed polygamous mating can be employed for all 4 strains studied.

The effects of type I collagenase on the degelification of chimpanzee (Pan troglodytes) semen plug and sperm quality

BackgroundSemen from the chimpanzee species becomes a colloidal solid after ejaculation. The formation of this copulatory plug is believed to prevent additional spermatozoa of subsequent mating events from accessing the ova. However, this naturally preserved strategy hampers the processes for sperm preparation. In this study, we investigated whether collagenase can be used to degelify the semen plug and accelerate the semen liquefaction process in zoo captive chimpanzee species (Pan troglodytes).ResultsWe showed that incubation of chimpanzee ejaculates with 0.1% type I collagenase efficiently and significantly (p < 0.05) releases 2.7-fold more spermatozoa from the coagulated ejaculates, and this degelification process did not alter sperm morphology or viability; nor did it stimulate spontaneous capacitation or an acrosome reaction as assessed by tyrosine phosphorylation and peanut agglutinin stains; moreover, based on computer assisted sperm analysis assay, motility-related parameters remained similar to those of untreated spermatozoa. When collagenase effects were evaluated on cryopreserved sperm samples, we observed post collagenase treatment in which 2.5% glycerol, as a cryoprotectant, preserved sperm acrosome integrity better than 7.8%; however, 7.8% glycerol, as a cryoprotectant, maintained sperm motility better than that of 2.5% glycerol.ConclusionsOur results demonstrated for the first time that type I collagenase can be used to obtain a significantly higher number of spermatozoa from colloid chimpanzee semen ejaculate without affecting the physiological properties of spermatozoa, and these results are critical for the subsequent gamete development. Our results would benefit sperm preparation processes and cryopreservation efficiency per ejaculate, as more unaffected spermatozoa can be released from the semen plug within a shorter period of time. These results would also benefit the genetic diversity of the chimpanzee species, using sperm cells from less dominant individuals, and for achieving better pregnancy success in primates with significantly higher amounts of sperm for artificial insemination.

Comparative female genital morphology in Stegodyphus spiders (Araneae: Eresidae)

The anatomy of the female genitalia sets the arena for sperm competition in species in which females mate multiply and store sperm. In spiders, females possess cuticular internal structures that have evolved into diverse sperm storage sites. Here, we investigate the female genital morphology of seven eresid spider species. We used X-ray micro-computed tomography for 3D reconstruction of the anatomy of the female genital system in the social Stegodyphus dumicola, S. mimosarum and S. sarasinorum, and the subsocial S lineatus, S. pacificus, S. tentoriicola as well as Eresus sandaliatus. We used histology to assess the sites of sperm storage in two selected species S. lineatus and S. dumicola. Our results show that the internal genitalia of the Stegodyphus species consist of two bilateral folds instead of closed ducts as often reported for entelegyne spiders. Along each fold, three regions are discriminable that differ in their specific morphology but are all surrounded by glandular tissue. Between species, the regions differ in shape and dimension. In virgin females, the lumina of all regions are filled with secretion. In mated females, spermatozoa were found in all three regions; however, only those sperm that are stored in the posterior region seem to become activated. Sperm found in anterior regions are embedded in a substance that might act as a mating plug. Our data suggest that the regions of the female genital tract differ in accessibility by the male as well as in their potential use for ejection and manipulation of sperm by the female.

Reproductive and developmental toxicity of 2- (2H-1, 2, 3-benzotriazol-2-yl) -4-methyl-6- (2-methylpropen-2-yl) phenol in mice: an experimental study

Objective: To investigate the reproductive and developmental toxicity of 2- (2H-1, 2, 3-benzotriazol-2-yl) -4-methyl-6- (2-methylpropen-2-yl) phenol in mice and to provide a basis for its risk assessment. Methods: The reproductive and developmental toxicity of 2- (2H-1, 2, 3-benzotriazol-2-yl) -4-methyl-6- (2-methylpropen-2-yl) phenol was tested using the screening method of chemicals with reproductive and developmental toxicity in "Chemical Testing Method" (SEPA). After five days of adaptive feeding, 120 specific pathogen-free healthy Kunming mice (male/female ratio=1:1) were orally administered 0 (control) , 146, 292, and 584 mg/kg 2- (2H-1, 2, 3-benzotriazol-2-yl) -4-methyl-6- (2-methylpropen-2-yl) phenol for two weeks. One male mouse was mated with one female mouse in a single cage. The day on which a vaginal plug was observed was defined as gestation day 0 (GD0). The exposure for female mice was sustained to four days postpartum and the exposure for male mice was sustained for two weeks after mating. The body weight, food intake, body length, tail length, and sex ratio were recorded and the reproductive index was calculated. The reproductive organs were weighed and subjected to histopathological examination. Results: The 584 mg/kg group had significantly lower body weight at weeks 5 and 6 and food intake at week 6 in male mice, uterus weight and uterus/body weight ratio in female mice, and body weight, body length, and tail length on day 0 in offspring compared with the control group (all P<0.05). The 292 mg/kg group had significantly lower testis weight of male mice and food intake of female mice at gestational week 2 than the control group (both P<0.05). The 146 mg/kg group had significantly lower food intake of female mice at gestational week 2 than the control group (P<0.05) . Conclusion: For male and female Kunming mice, the no observed adverse effect levels of 2- (2H-1, 2, 3, -benzotriazol-2-yl) -4-methyl-6- (2-methylpropen-2-yl) phenol are both 146 mg/kg.

Effects of ciprofloxacin and quercetin on fetal brain development: a biochemical and histopathological study

Purpose Teratogens cause birth defects and malformations while human development is being completed. In pregnancy, urinary tract infection (UTI) is a common health problem caused by bacteria. The fluoroquinolones such as ciprofloxacin, levofloxacin, moxifloxacin, and gemifloxacin can treat various types of bacterial infections successfully. The aim of this study is to determine whether the use of ciprofloxacin during pregnancy causes oxidative stress on brain tissues of the fetus, and whether quercetin contributes to prevent this damage if stress has already occurred. Materials and methods In our study, 22 young female Wistar albino rats weighing 250 g were used. Rats were mated overnight in separate plastic cages. Female rats were regarded as pregnant when a vaginal plug was observed, and these were divided into four groups of control, ciprofloxacin, quercetin, and cipro + quercetin. Two daily i.p. 20 mg/kg doses of ciprofloxacin were administered to ciprofloxacin group between 7 and 17 d of pregnancy. Throughout the study, daily (20 d) 20 mg/kg quercetin was dissolved in corn oil and administered to the quercetin group by oral gavage. Rats were fed ad libitum throughout the study. Fetuses were taken by C-section on the 20th day of pregnancy. Thereafter, the brain tissues were subjected to histological assessments and biochemical analyzes. Results The experimental groups were compared with the control group; ciprofloxacin affected fetal development, especially caused damage to neurons in brain tissue and cause hemorrhagic defects. And also, it was determined that many parameters were affected such as antioxidant parameters, enzyme levels and levels of malondialdehyde (MDA) (a marker of lipid peroxidation). Quercetin is a member of flavonoid with strong antioxidant properties, and our results indicate that the use of ciprofloxacin in pregnancy can result damage to fetal brain tissue. Conclusions Unlike these results when some parameters are evaluated it is understood that this harmful effects suppressed by quercetin.

Rescue of the fetal damage associated with high intrauterine pressure by 17β-estradiol injection in ovariectomized progesterone-treated pregnant mice.

The present study examined the effects of progesterone (P) and 17β-estradiol (E2) on fetal damage and intrauterine pressure in ovariectomized pregnant mice. The mice were ovariectomized on gestational day (GD) 9 (copulation plug = GD 0), and daily subcutaneous injection of various doses of P (2, 3 or 4 mg) or 4 mg P plus E2 (0.05 or 0.1 μg) was given thereafter. Although P alone increased percentage of normal fetuses on GD 17 dose-dependently, fetal injury with edematous hematomata on their extremities was frequently observed. In the group treated with 4 mg P, the injured fetus was found at the highest percentage (18%) and intrauterine pressure was significantly higher than that in intact pregnant mice (controls). No injured fetus on GD 17 was found by the treatment with 4 mg P plus 0.05 or 0.1 μg E2, and the treatments decreased the intrauterine pressure to the level of controls. Percentage of normal fetuses in the ovariectomized mice treated with 4 mg P plus 0.05 μg E2 was similar to that of controls, while that in the ovariectomized mice treated with 4 mg P plus 0.1 μg E2 markedly decreased. The results suggest that estrogen decreases intrauterine pressure to defend fetal damage in ovariectomized P-treated mice, and a high estrogen level interrupted pregnancy while keeping this estrogen action.

596: Ethyl pyruvate: A novel approach to protect the fetus against antenatal inflammatory damage

In pregnancies complicated by inflammation, activation of RAGE signaling pathways may be responsible for poor fetal and neonatal outcomes including stillbirth and neonatal brain injury. Ethyl pyruvate (EP) improved survival in murine models of endotoxemia via RAGE inhibition, and was already shown to be safe in humans. We investigated the effect of maternal administration of EP on neonatal growth and neurodevelopmental outcome using a genetically engineered over-expressing RAGETg murine model of in-utero inflammation. Saline or endotoxin (LPS, 5 or 10 μg/dam) were given intra-peritoneal (i.p.) on day 15 (d0=sperm plug), in mice randomly assigned to the following groups: 1) RAGETg and 2) Wild-type (WT) (n=9-30 pups/group) as controls. EP (40 mg/kg) was administered i.p. 2 hours later. Doses were chosen based on prior experiments demonstrating live birth. Pups underwent post-natal daily behavioral testing (i.e. cliff aversion, forelimb grasp, negative geotaxis) from birth to day 14 of life. Using immunohistochemistry (IHC), we assessed cortical neuronal density in animals sacrificed on day 20 of life. 1) In-utero exposure to EP did not increase post-natal mortality rate; 2) Compared to control, weight gain was inhibited in WT receiving LPS5 (p=0.02), which was ameliorated by EP (p=0.0001); 3) WT and RAGETg mice exposed to LPS demonstrated delay in achieving behavioral milestones, an effect reversed by EP (p<0.001, Figure); 4) WT and RAGETg mice showed improved neurobehavioral battery with EP independent of fetal gender; 5) EP protected against LPS-induced cortical neuronal loss (p=0.001) by IHC. We report a novel neuroprotective effect of EP from perinatal inflammatory insult.

Evaluating the potential of stichopus horrens selenka as a source of aphrodisiac agent on mice

Stichopus horrens Selenka or locally known as ‘gamat emas ’has been reputed to have aphrodisiac property which able to increase sexual desire and help to improve potency. However, there is no scientific evidence to support the claim. Therefore, this study was conducted to evaluate the potential ofS. horrens extract as a source of aphrodisiac agent. Three doses; 50, 100 and 200 mg kg ’ ’of S. horrens extract were administered intraperitoneally to male mice for measuring the mounting behavior and mating assessment while 200 mg kg−1 being administered for serum testosterone analysis. The effect of extract dose between each hour interval showed significant differences (p <0.05) on mounting frequency. Meanwhile, the treated mice show marked evidences in mounting behavior at all doses level although only mild aphrodisiac effect was shown at dose 50 mg kg−1. Despite this, the extract at doses 100 and 200 mg kg−1 showed significant difference (p <0.05) compared to negative control. This indicated the potentiality ofS. horrens extract to pose aphrodisiac property and be as competence as sildenafil citrate. In addition, there was an increased level of serum testosterone in treated male mice at dose 200 mgkg−1 compared to negative control. In mating assessment test, no sperm plug was observed on female mice. This study provided a preliminary evidence on the potential of S. horrens as a source of aphrodisiac agent.

Arsenic aggravated reproductive toxicity in male rats exposed to lead during the perinatal period.

The aim of this study was to assess the reproductive toxic effects of arsenic on adult Wistar rats exposed to lead during the perinatal period. The pregnant rats were allowed ad libitum access to tap water containing 819 mg of lead (Pb) per L or without Pb from conception until weaning. Litter size, survival rate and developmental milestones of the pups delivered by Pb exposed dams were comparable to those of the control rats. Conversely, the pups exposed to Pb during the perinatal period exhibited significant delay in cliff avoidance, negative geotaxis, surface righting reflex, ascending wire mesh and testis descent. The control and perinatal Pb-exposed male rats were maintained on tap water containing 2.3 mg of arsenite (As) per L or without arsenite from the pubertal period (post-natal day 55) to adulthood (post-natal day 115) and assessed for reproductive end points. The results revealed that the (1) relative weights of the testis, epididymis, seminiferous tubules and ventral prostate; (2) daily sperm production; (3) epididymal sperm density and (4) numbers of motile, viable, and HOS tail swelled sperm declined significantly in the rats exposed to either Pb or As. The activity levels of testicular 3β- and 17β-hydroxysteroid dehydrogenases were also significantly decreased in the experimental rats. Significant elevation in the levels of reactive oxygen species and lipid peroxidation in association with reduced activities of antioxidant enzymes in the testis and different epididymal regions was recorded in the experimental rats. In the fertility study, although each male in the control and experimental groups produced a copulatory plug and impregnated a female, the mean conception time significantly increased in the experimental groups. The mean number of implantations decreased significantly in the females mated with the experimental males. Moreover, the results of the present study also indicate that reproductive alterations were more deteriorated in the Pb-exposed rats treated with arsenic when compared to individual exposures. In conclusion, the data clearly suggest that reproductive toxicity in male rats exposed to Pb during the perinatal period is exacerbated by As treatment during the pubertal period.