Evaluation of the aquaporin molecules characterization in the sperm cells of men from different aged.

Background/aim: Male infertility rises for many reasons, along with age; therefore, we aimed to research the characterization of aquaporin-3, 7, and 8 in human sperm belonging to different age groups. Material and methods: This study was conducted on sperm samples of men aged over 18 years. A total of 60 men were included in the study and divided into three age groups: group 1, age 18-25 years (n = 20); group 2, age 26-35 years (n = 20); and group 3, age ≥35 years (n = 20). Sperm ejaculates obtained from each participant were used for spermiogram tests, Kruger strict morphology analysis, and immunohistochemistry. Results: We observed no statistically significant differences in terms of macroscopic and microscopic sperm testing. The immunostaining score of aquaporin-3 was the lowest in group 1 and increased in group 3 and group 2, respectively (p < 0.05). Aquaporin-8 immunostaining only increased in group 2 (p < 0.05). Aquaporin-7 immunostaining scores were not different between the groups (p > 0.05). When the immunostaining scores of aquaporin molecules were compared with each other, aquaporin-7 was significantly increased compared with the others (p < 0.05). Conclusion: According to the results, it can be stated that aquaporin-3 and aquaporin-8 molecules were more expressed at age 26 to 35 years, and aquaporin-7 was densely expressed from age 18 to 25 years. If the characterization of these molecules is adversely affected, male infertility may eventually emerge. We recommend further advanced-level studies on this subject.

CREB1 and PPAR-α/γ Pathways in Hepatic Ischemia/Reperfusion: Route for Curcumin to Hepatoprotection.

Background: Hepatic ischemia/reperfusion injury is a major problem that can exacerbate complications, particularly in liver transplantations. Objectives: This study aimed to investigate the cellular mechanisms of ischemia/reperfusion injury and hepatoprotection by curcumin. Methods: Wistar albino rats were divided into four groups as Control, Sham, I/R, and Cur+I/R. Hepatic ischemia/reperfusion was induced in I/R and Cur+I/R animals, the latter of which was also given 50 mg/kg/day of curcumin for 14 days. Liver aminotransferases and the transcription regulators of inflammation (RelA, IκB, PPAR-α, PPAR-γ, CREB1) were examined along with the histological examination. Results: Hepatic ischemia/reperfusion was found to disrupt hepatic microstructure and downregulate PPAR-α, PPAR-γ, and CREB1 transcripts. Curcumin supplementation in hepatic ischemia/reperfusion recovered the structural organization and promoted the hepatocyte regeneration while increasing expressions of PPARs and CREB1. RelA and IκB were found unaltered, possibly due to the crosstalk between targeted transcripts by ischemia/reperfusion and curcumin. Conclusions: In sum, PPAR-α/γ and CREB1 were involved in hepatic ischemia/reperfusion and, moreover, were detected to be stimulated by curcumin. PPAR and CREB pathways were found to provide a route to hepatoprotection for curcumin supplementation as evidenced by the microstructural improvement.

Adverse effects of oxytetracycline and enrofloxacin on the fertility of Saanen bucks.

This study aimed to determine the adverse effects of oxytetracycline and enrofloxacin application on the fertility of Saanen bucks. For this purpose, twenty-four bucks were divided into three groups. Group I (control group) received only 5 ml of 0.9% NaCl for 7 days, group II was given a single dose of 20 mg/kg oxytetracycline and group III was given at a dose of 2.5 mg/kg per day for 7 days intramuscularly. Serum and semen samples were collected from the bucks at post-treatment 1, 3, 5, 7, and 9 days and examined spermatological parameters (quantity, motility, density, abnormal sperm ratio, and live-dead sperm ratio), serum testosterone levels (with ELISA) and sperm DNA parameters (with Comet assay). The results showed no change in sperm volume, abnormal sperm rate, and dead-live sperm ratio in group II and III following oxytetracycline and enrofloxacin administration. However, a decrease in sperm density, sperm motility, mass activity, and testosterone levels, and an increase in sperm DNA damage were detected. These spermatological parameters (density, motility, mass activity) and testosterone levels were less decreased and sperm DNA damage was less increased in group II than group III. The greater damage in group III may be attributed to the longer duration of enrofloxacin administration compared to oxytetracycline and the effect of enrofloxacin on DNA. The results obtained from this study suggest that usage of oxytetracycline and especially enrofloxacin should be restricted and antibiotics with fewer side effects on sperm should be preferred in Saanen bucks during the reproduction period.

Prenatal exposure to low-dose diclofenac sodium does not affect total neuron numbers in spinal segment T13 in rats.

The main purpose of the present study was to investigate the effects of diclofenac sodium (DS) on the total number of neurons in segment T13 of the spinal cord of offspring of pregnant rats using stereological methods. Eighteen adult female Wistar albino rats weighing 150-200g were used. Pregnant female rats were divided into three groups; a control group, a sham group and a DS (1mg/kg, intramuscular) exposed group. The DS and sham groups received injection from the 5th day of gestation to the 19th. Twenty eight days after birth, the offspring rats were perfused with 4% buffered formalin. T13, which is one of transverse spinal cord segments, were isolated and processed for routine paraffin histology. 5µm sections were obtained using a rotary microtome according to systematic random sampling strategies. Every 40th section was taken and sections were stained with modified Giemsa. All types of motor neuron cell were identified according to their morphology. In this study, the "disector-Cavalieri combination" method was used in the stereological examination of neurons. The motor neurons were counted in the right gray matter of the ventral horn in the spinal cord segment. The Kruskal-Wallis test was used for comparison the groups. In terms of motoneuron number, no significant difference among the groups was found (p>0.05). In conclusion, our results indicated that prenatal exposure to DS has no effect on the total number of motor neuron of the offspring rats.

The acute effects of thymoquinone on acute peripheral nerve injury: an experimental study.

BACKGROUND: The purpose of this study was to evaluate the acute effects of thymoquinone (TQ) on acute nerve injury. METHODS: A rat model of crush injury of the sciatic nerve was used. Animals were divided into 3 groups: control, trauma, and TQ treatment groups (n=6 per group). Seven days after injury, sciatic nerve specimens were obtained from the site of the injury and analyzed histologically and stereologically. Axon diameter, myelin thickness, and axon density were measured. RESULTS: There were no significant differences in axon diameter, myelin thickness, or axon density among groups. CONCLUSION: TQ has no acute therapeutic effect on acute nerve injury.

Enhanced G2/M Arrest, Caspase Related Apoptosis and Reduced E-Cadherin Dependent Intercellular Adhesion by Trabectedin in Prostate Cancer Stem Cells.

Trabectedin (Yondelis, ET-743) is a marine-derived tetrahydroisoquinoline alkaloid. It is originally derived from the Caribbean marine tunicate Ecteinascidia turbinata and currently produced synthetically. Trabectedin is active against a variety of tumor cell lines growing in culture. The present study focused on the effect of trabectedin in cell proliferation, cell cycle progression, apoptosis and spheroid formation in prostate cancer stem cells (CSCs). Cluster of differentiation (CD) 133+high/CD44+high prostate CSCs were isolated from the DU145 and PC-3 human prostate cancer cell line through flow cytometry. We studied the growth-inhibitory effects of trabectedin and its molecular mechanisms on human prostate CSCs and non-CSCs. DU-145 and PC-3 CSCs were treated with 0.1, 1, 10 and 100 nM trabectedin for 24, 48 and 72 h and the growth inhibition rates were examined using the sphere-forming assay. Annexin-V assay and immunofluorescence analyses were performed for the detection of the cell death. Concentration-dependent effects of trabectedin on the cell cycle were also evaluated. The cells were exposed to the different doses of trabectedin for 24, 48 and 72 h to evaluate the effect of trabectedin on the number and diameter of spheroids. According to the results, trabectedin induced cytotoxicity and apoptosis at the IC50 dose, resulting in a significant increase expression of caspase-3, caspase-8, caspase-9, p53 and decrease expression of bcl-2 in dose-dependent manner. Cell cycle analyses revealed that trabectedin induces dose-dependent G2/M-phase cell cycle arrest, particularly at high-dose treatments. Three-dimensional culture studies showed that trabectedin reduced the number and diameter of spheroids of DU145 and PC3 CSCs. Furthermore, we have found that trabectedin disrupted cell-cell interactions via E-cadherin in prostasphere of DU-145 and PC-3 CSCs. Our results showed that trabectedin inhibits cellular proliferation and accelerates apoptotic events in prostate CSCs; and may be a potential effective therapeutic agent against prostate cancer.

Tissue damage in kidney, adrenal glands and diaphragm following extracorporeal shock wave lithotripsy.

This study was designed to investigate whether exposure to short-term extracorporeal shock wave lithotripsy (ESWL) produces histologic changes or induces apoptosis in the kidney, adrenal glands or diaphragm muscle in rats. The effect of shock waves on the kidney of male Wistar rats (n = 12) was investigated in an experimental setting using a special ESWL device. Animals were killed at 72 h after the last ESWL, and the tissues were stained with an in situ Cell Death Detection Kit, Fluorescein. Microscopic examination was performed by fluorescent microscopy. Apoptotic cell deaths in the renal tissue were not observed in the control group under fluorescent microscopy. In the ESWL group, local apoptotic changes were observed in the kidney in the area where the shock wave was focused. The apoptotic cell deaths observed in the adrenal gland of the control group were similar to those observed in the ESWL groups, and apoptosis was occasionally observed around the capsular structure. Apoptotic cell deaths in the diaphragm muscle were infrequently observed in the control group. Apoptosis in the ESWL group was limited to the mesothelial cells. This study demonstrated that serious kidney, adrenal gland and diaphragm muscles damage occurred following ESWL, which necessitated the removal of the organ in the rat model. It is recognized that the ESWL complications related to the kidney, adrenal gland and diaphragm muscles are rare and may be managed conservatively.

A histological investigation concerning the effects of diclofenac sodium to the lung in 4- and 20-week-old rats treated prenatally.

OBJECTIVE: We aimed to investigate the possible postnatal effects on the lung tissues of the rat offspring treated with diclofenac sodium (DS) during pregnancy. METHODS: After mating, pregnant female rats were separated into the control (n = 10) and DS (n = 10) groups. DS (1 mg/kg) was injected intraperitoneally (i.p) to the drug-treated group for the period of gestational days 5-19. Physiological saline (1 ml, i.p.) was given to the control groups. After birth, pups were separated into DS treatment groups (n = 24) and control group (n = 24). The DS and control group animals were anaesthetised with i.p. injection of urethane and their lungs were removed to prepare for histopathological evaluation. RESULTS: Histological examination of the lung tissues of the 4- and 20-week-old rats revealed no significant differences between males and females in both the control and DS treated rats. CONCLUSION: Because of the use of DS in the pregnant women further studies are needed in this field.