First comprehensive report of bacteria spp. associated with cloaca of Laudakia nupta (Sauria: Agamidae) in Iran using molecular studies / 한국실험동물학회지

Iran bears a remarkable variety of reptiles. One of the lizard families occurring in Iran is the Family Agamidae which is widely are distributed throughout the old world. The large-scaled rock agamid, Laudakia nupta, is one of the well-known agamid. There are few reports of cloacal microbial on reptiles hence their function in cloacae remains unknown. Laudakia nupta usually live in rural and urban areas and close vicinity to man, they are likely to play an important role in the spread of disease that may be caused by these microorganisms and their transmission to man. Therefore, the aim of this study was to identify the bacterial flora colonizing the cloacal region of Laudakia nupta using molecular studies. The cloacal fluids were directly placed on nutrient agar (NA) plates and incubated at 25 ± 2 ℃ for 48 h. The resulting bacterial colonies were transferred to fresh nutrient agar (NA) plates for molecular studies. Twelve isolates were obtained from 17 specimens of Laudakia nupta. All bacteria isolates were identified as Bacillus subtillis (5), Bacillus cereus (4), Bacillus sp. (1), Pseudomonas putida (1), and Pseudomonas sp. (1) based on partial sequences of the 16 s rRNA gene. This is the first comprehensive report of bacteria spp. associated with cloaca of Laudakia nupta using molecular studies. In this research, we found that Laudakia nupta can be a carrier of bacteria which can transfer microorganisms to hosts.

Ovary stereological features and serum biochemical factors following induction of polycystic ovary syndrome with testosterone enanthate in mice: an experimental study

Background: Polycystic ovary syndrome [PCOS] is an endocrine disorder featured by insulin resistance and hyperandrogenism. Testosterone enanthate can induce PCOS in mice models

Objective: We investigated the ovary stereological features along with the oxidative stress and inflammatory factors in mice following PCOS induction using testosterone enanthate

Materials and Methods: Twelve female NMRI mice [3 wk old] were divided into 2 groups [n=6/each]: Control and PCOS. PCOS was induced through daily injections of testosterone enanthate [1 mg/100g subcutaneous s.c for 5 wk]. Finally, ovaries were studied stereologically. The serum levels of the follicle-stimulating hormone, luteinizing hormone, testosterone, interleukin-6, and tumor necrosis factor-alpha were measured using ELISA kit. Serum levels of Malondialdehyde and the antioxidant capacity were measured relatively using thiobarbituric acid and ferric reducing antioxidant power assay

Results: The mean total volume of ovary and the mean volume of cortex [p<0.001], volume of oocyte in the preantral [p=0.011] and antral follicle [p=0.015], thickness of zona pellucida [p=0.016], the number of antral follicles [p=0.012], the serum levels of follicle-stimulating hormone [p<0.001] and the antioxidant capacity [p=0.020] reduced significantly in the PCOS group compared to the control. The number of primary [p=0.017] and preantral [p=0.006] follicles and the serum levels of testosterone [p<0.001], Luteinizing hormone [p=0.002], Malondialdehyde, Interleukin 6 and Tumor necrosis factor-? [p<0.001] showed a significant increase in the PCOS group compared to the control

Conclusion: Testosterone enanthate induced PCOS causes stereological features in the ovary, increases the oxidative stress and inflammatory markers in mice

Ethanolic extract of Ficuscarica leave Suppresses Angiogenesis by Regulating VEGF-A and Integrin β3 mRNA Expression in Human umbilical vein endothelial cells.

Background: In the present study, we investigated the anti-angiogenic effects of the ethanol extract of Ficus carica leave on human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were used in this study. The cells were cultured in DMEM medium and then incubated with different concentrations of ethanolic extract of Ficus carica leave (0-25 μg\ml) in the presence or absence of the extract for 24 hours. Cell viability was analyzed using neutral red assay. Endothelial cell tube formation was measured with the Matrigel basement membrane matrix. The level of VEGF and Integrin β3 mRNA expression in the HUVECs was measured with reverse-transcription quantitative real-time polymerase chain reaction (RT-q real time PCR). Results: We observed that the extract dose dependently inhibited the tube formation of HUVECs. Furthermore, the extract significantly decreased mRNA expression levels of VEGF-A and Integrin β3 in HUVECs at 20 μg\ml concentration of the extract compared to untreated control cells (P<0.05). Conclusion: Our findings suggest that ethanolic extract of Ficus carica leave contains anti-angiogenic activities and could be a candidate as a potential agent for the prevention of angiogenesis related disorders.

Hydrostatic pressure affects in vitro maturation of oocytes and follicles and increases granulosa cell death

This study examines the effects of hydrostatic pressure on in vitro maturation [IVM] of oocytes derived from in vitro grown follicles. In this experimental study, preantral follicles were isolated from 12-day-old female NMRI mice. Each follicle was cultured individually in Alpha Minimal Essential Medium [alpha-MEM] under mineral oil for 12 days. Then, follicles were induced for IVM and divided into two groups, control and experiment. In the experiment group follicles were subjected to 20 mmHg pressure for 30 minutes and cultured for 24-48 hours. We assessed for viability and IVM of the oocytes. The percentage of apoptosis in cumulus cells was determined by the TUNEL assay. A comparison between groups was made using the student's t test. The percentage of metaphase II oocytes [MII] increased in hydrostatic pressure-treated follicles compared to controls [p<0.05]. Cumulus cell viability reduced in hydrostatic pressure-treated follicles compared to controls [p<0.05]. Exposure of follicles to pressure increased apoptosis in cumulus cells compared to controls [p<0.05]. Hydrostatic pressure, by inducing apoptosis in cumulus cells, participates in the cumulus oocyte coupled relationship with oocyte maturation

Hydrostatic pressure improves in-vitro maturation of oocytes derived from vitrified-warmed mouse ovaries

Cryopreservation has limited successes and in-vitro maturation is used to improve its results. Hydrostatic pressure [HP] plays an important role in follicular development. This study was designed to examine the effects of HP on in-vitro maturation of oocytes and cell death in cumulus cells derived from vitrified-warmed mouse ovaries. Preovulatory follicles were harvested from non-vitrified and vitrified-warmed 6-8 week-old female NMRI mouse ovaries and randomly assigned to following groups: non-vitrified [control], non-vitrified with HP exposure [treatment I], vitrified-warmed [treatment II] and vitrified-warmed with HP exposure [treatment III]. The follicles of treatments I and III were subjected to HP [20 mmHg] for 30 min and after that all groups were cultured for 24h and assessed for in-vitro maturation of oocytes. The viability and apoptosis of cumulus cells and oocytes were assessed using supravital nuclear staining and TUNEL assay, respectively. Oocytes harvested follicles in both control and treatment II had a significantly lower percentage of metaphase II oocytes [MII] than the treatment I and III [23.5 +/- 3.1, 15.03 +/- 4.6 and 32.7 +/- 3.2, 25.5 +/- 4.6; respectively] [p<0.05]. Viability of the cumulus cells reduced in treatment I, II and III [83.4, 83.3 and 77.7%] compared to control [86.9%], [p<0.05]. The apoptotic index in cumulus and oocyte complexes in treatments I and III [10.7 +/- 0.8 and 15.3 +/- 0.8] was higher than in control and treatment II [6.7 +/- 0.5 and 9.7 +/- 0.5] [p<0.05]. These results demonstrate that HP had a mild effect on cell death incidence in cumulus cells without any effect on oocyte. However, it can be used as a mechanical force to improve in-vitro maturation of oocytes derived from vitrified-warmed mouse ovaries