Expression and localization of insulin-like growth factor gene family members in the caprine ovarian and uterine tissues during different pregnancy stages.

Goats are of significant economic importance, yet our knowledge of the molecular pathways involved in their pregnancy remains limited. This study aims to investigate the role of IGFs in uterine and ovarian cellular events during pregnancy in goats. Forty-two Hair Goats were examined, including four pregnancy groups representing embryo-positive (G1, n=7), early (G2, n=7), mid (G3, n=7), and late pregnancy (G4, n=7), as well as two luteal stage groups representing early (G5, n=7) and late (G6, n=7) phases. Uterine and ovarian tissues were collected, and RT-qPCR and immunohistochemistry were performed to evaluate IGF expression. The results showed that IGF1 and IGF2 expressions were significantly higher in G1 than in other pregnancy and control groups (p < 0.05). Additionally, IGFBP1 expression was higher in G2 than in G1 and G4 (p < 0.05), and IGFBP3 expression was higher in G4 than in any other pregnancy stage (p < 0.05). However, no statistically significant differences were observed in the expression levels of IGFBP4 and IGFBP6 between any of the groups. Finally, IGFBP5 expression was significantly higher in G1, G3, and G4 compared to G2 (p < 0.05). Overall, the dynamic changes observed in the expression of the IGF gene family during different stages of pregnancy highlight the crucial role of IGFs in regulating pregnancy in goats.

The curative effect of ozonated bidistilled water on Escherichia coli-induced endometritis in rats.

BACKGROUND: Endometritis is a prevalent and challenging condition that affects livestock, and its effective treatment is the use of intrauterine antibiotics. Antibiotic use may lead to negative consequences, including residue in tissues and the emergence of antibiotic resistance. OBJECTIVES: The aim of this study was to assess the therapeutic potential of ozonated bidistilled water in the treatment of Escherichia coli-induced endometritis in rat models, comparing it to conventional antibiotic treatment. METHODS: A total of 21 female Wistar Albino rats were used for this study. E. coli from the uterus of cows with endometritis at a concentration of 1.0 × 109 cfu/mL inoculated into rat uteruses. After inoculation, it was confirmed macroscopically that experimental endometritis was induced in all individuals. And, the rats with endometritis were randomly divided into three groups treated with intrauterine bidistilled water enriched with ozone, antibiotics and 0.9% NaCl. The uterine tissues were examined histopathologically. RESULTS: In the semiquantitative analyses, significantly lower histopathological scores were observed for both the bidistilled water enriched with ozone and antibiotic-treated endometritis groups compared to the control group (p < 0.05). CONCLUSIONS: Ozone-enriched bidistilled water can be used in the treatment of experimentally induced endometritis in rats, and can provide effective microbiological and histopathological improvement. Further studies involving larger populations of different species are needed to determine the medical suitability of this new treatment.

Expression of the insulin-like growth factor (IGF) gene family in feline uterus during pregnancy.

Members of the IGF gene family participate in cell differentiation and proliferation during pregnancy. We used 35 cats assigned to experimental groups (G) based on pregnancy stages: G1, pre-implantation; G2, implantation; G3, early pregnancy; G4, mid-pregnancy; G5, nonpregnant. Quantitative polymerase chain reaction (qPCR) and immunohistochemistry were used to analyze the expression of the IGF gene family. During pregnancy, expression of IGF-1 gene was significantly greater at implantation sites in the G1 and G2 groups than at placentation sites in G3 and G4 groups. IGF-2 expression was greater in the G2, G3 and G4 groups than in G1. Expression of the IGF-1R gene was significantly greater at placental sites in G3 than in G1 and G4. IGF-2R genes were expressed in all groups. Insulin-like growth factor binding proteins (IGFBPs) were expressed at intensities that depended on the stage of pregnancy; they were detected in different cell types and at different sites in the uterus. We found that members of the IGF gene family were expressed differentially in the endometrium during pregnancy. Our findings suggest that the IGF family may be a regulatory factor for pregnancy in cats.

Gene expression profiles of some cytokines, growth factors, receptors, and enzymes (GM-CSF, IFNγ, MMP-2, IGF-II, EGF, TGF-ß, IGF-IIR) during pregnancy in the cat uterus.

Early pregnancy is one of the most critical periods of pregnancy, and many factors such as cytokines, enzymes, and members of the immune system have to cooperate in a balanced way. In the present study, the gene expression profiles of factors associated with pregnancy such as EGF, transforming growth factor beta, granulocyte-macrophage colony-stimulating factor, interferon gamma, insulin-like growth factor 2, insulin-like growth factor 2 receptor, and matrix metalloproteinase 2 were analyzed in uterine tissues of female cats. The cats were assigned to five groups: G1 (embryo positive, n = 7; 7th day after mating), G2 (after implantation, n = 7; 20th day after mating), G3 (midgestation, n = 7; 24-25th day after mating), G4 (late gestation, n = 7; 30-45th day after mating), G5 (oocyte group, n = 7; 7th day after estrus). Tissue samples from the uterus and placenta were collected after ovariohysterectomy. Relative messenger RNA levels were determined by real-time polymerase chain reaction. All the factors examined were detected in all tissue samples. In the course of pregnancy, significantly higher expression of EGF and matrix metalloproteinase 2 in G2 than in G1 was observed (P < 0.05). Insulin-like growth factor 2 expression was higher in all groups than in G1 (P < 0.05). Upregulation of EGF during implantation was detected. The expression of interferon gamma was significantly higher in G3 than in G1 (P < 0.05). Transforming growth factor beta and granulocyte-macrophage colony-stimulating factor were constantly expressed in all groups. In conclusion, the expressions of these factors in feline uterine tissue at different stages of pregnancy might indicate that these factors play roles in the development of pregnancy such as trophoblast invasion, vascularization, implantation, and placentation.

Expression of hypoxia-inducible factors and vascular endothelial growth factor during pregnancy in the feline uterus.

Hypoxia-inducible factors (HIFs) and vascular endothelial growth factor (VEGF) have critical roles during the development of the fetomaternal unit. The HIFs regulate placentation and vascularization by stimulation of VEGF gene expression. This study aimed to investigate the expression profiles of HIF gene family and VEGF in the cat uterus during pregnancy. Tissue samples of the whole uterine wall were collected after ovariohysterectomy and allocated to the following groups: embryo positive (group 1 [G1], n = 7, 7 days after mating), early pregnancy (group 2 [G2], n = 7, 20 days after mating), mid-pregnancy (group 3 [G3], n = 7, 24 days after mating), late pregnancy (group 4 [G4], n = 7, 30-45 days after mating), and oocyte positive groups (group 5 [G5], n = 7, 7 days after induction of ovulation with GnRH analog). Relative mRNA levels were determined by real-time polymerase chain reaction. As housekeeping gene, glyceraldehyde-3-phosphate dehydrogenase was used. The relative gene expression of HIF1A in G5 was found to be significantly higher than that of other groups (G1, G2, G3, and G4) (P < 0.05). In addition, the expression of HIF2A in G5 was higher than that of G1 and HIF2A gene expression at placentation sites of G4 was higher than in G1, G2, and G3 (P < 0.05). Immunohistochemistry indicated that HIF1A, HIF2A, and VEGF expressions were observed in different cell types of uterine and placental tissues in late pregnancy and oocyte groups. The expression of HIF3A did not change significantly in any group investigated. These observations suggest that HIFs and VEGF may play a role in the establishment and development of pregnancy.

Tarantula cubensis extract alters the degree of apoptosis and mitosis in canine mammary adenocarcinomas.

In the present study, 13 clinical cases of canine mammary adenocarcinoma were evaluated in order to understand the effect of Tarantula cubensis extract (TCE) on tumor tissue. Punch biopsies were taken from the tumors before treatment with TCE. Subcutaneous injections of TCE were administered three times at weekly intervals (3 mL per dog). Between days 7 and 10 after the third injection, the tumor masses were extirpated by complete unilateral mastectomy. Pre- and post-treatment tumor tissues were immunohistochemically assessed. The expression of B-cell lymphoma 2 (Bcl-2) was found to be higher in pre-treatment compared to post-treatment tissues (p < 0.01) whereas Ki-67 expression was lower in post-treatment tissues (p < 0.01). No significant differences in fibroblast growth factor or vascular endothelial growth factor expression were observed between pre- and post-treatment tissues (p > 0.05). The apoptotic index was determined to be low before treatment and increased during treatment. These results suggest that TCE may be effective for controlling the local growth of canine mammary adenocarcinoma by regulating apoptosis.

Ovarian response and embryo yield of Angora and Kilis goats given the day 0 protocol for superovulation in the non-breeding season.

The aim of this study was to compare ovarian response and embryo yield of Day 0 protocol in Angora goats (AG) and indigenous Kilis goats (KG) in the non-breeding season. A total of 16 Angora goats (AG group) and 11 Kilis goats (KG group) were used in this study. In the synchronization process, after controlled internal drug release withdrawal, when estrus signs were observed, natural mating was performed. Ovarian response was determined by synchronized laparotomy 6 days after natural mating, and number of corpora lutea (CL) was recorded. Embryos were collected and morphologically evaluated by stereomicroscope. Synchronization rates did not differ between AG (88%, 14/16) and KG group (91%, 10/11). In AG and KG groups, the proportion of CL on the right (44% and 53%, respectively) and left (56% and 47%, respectively) ovaries were similar. The CL number per animal did not differ significantly between the two breeds and was determined as 4.4 ± 0.90 in AG group and 6.4 ± 1.44 in KG group. Transferable embryo yields were significantly higher in AG group (31/42, 74%) compared to KG group (16/46, 35%) in the non-breeding season (P < 0.01). In conclusion, it is suggested that the day 0 protocol can be used for goat superovulation in the non-breeding season; however, transferable embryo yields are affected by the breed.

Time related changes in luteal prostaglandin synthesis and steroidogenic capacity during pregnancy, normal and antiprogestin induced luteolysis in the bitch.

In nonpregnant and pregnant dogs the corpora lutea (CL) are the only source of progesterone (P4) which shows an almost identical secretion pattern until the rapid decrease of P4 prior to parturition. For the nonpregnant dog clear evidence has been obtained that physiological luteal regression is devoid of a functional role of the PGF2alpha-system and seems to depend on the provision of StAR. Yet in pregnant dogs the rapid prepartal luteal regression, coinciding with an increase of PGF2alpha, may be indicative for different regulatory mechanisms. To assess this situation and by applying semi-quantitative Real Time (Taq Man) RT-PCR, expression patterns were determined for the following factors in CL of pregnant and prepartal dogs and of mid-pregnant dogs treated with the antiprogestin Aglepristone: cyclooxygenase 2 (Cox2), prostaglandin E2 synthase (PGES), prostaglandin F2alpha synthase (PGFS), its receptors (EP2, EP4 an FP), the steroidogenic acute regulatory protein (StAR), 3beta-hydroxysteroid-dehydrogenase (3betaHSD) and the progesterone receptor (PR). Peripheral plasma P4 concentrations were determined by RIA. CL were collected via ovariohysterectomy from pregnant bitches (n=3-5) on days 8-12 (Group 1, pre-implantation period), days 18-25 (Group 2, post-implantation period), days 35-40 (Group 3, mid-gestation period) and during the prepartal progesterone decline (Group 4). Additionally, CL were obtained from groups of 5 mid-pregnant dogs (days 40-45) 24h, respectively 72h after the second treatment with Aglepristone. Expression of Cox2 and PGES was highest during the pre-implantation period, that of PGFS and FP during the post-implantation period. EP4 and EP2 revealed a constant expression pattern throughout pregnancy with a prepartal upregulation of EP2. 3betaHSD and StAR decreased significantly from the pre-implatation period to prepartal luteolysis, it was matched by the course of P4 concentrations. Expression of the PR was higher during mid-gestation and prepartal luteolysis than in the two preceding periods. After application of Aglepristone the overall mRNA-expression resembled the situation during prepartal luteolysis except for EP2, which remained unchanged. These data suggest that - as in the nonpregnant bitch - also in the pregnant bitch luteal production of prostaglandins is associated with luteal support rather than luteolysis. On the other hand induction of luteolysis by the PR blocker Aglepristone points to a role of luteal P4 as an autocrine factor in a positive loop feedback system controlling the availability of P4, StAR and 3betaHSD.