The effect of protocatechuic acid on ovarian histopathology and reserve in rat ovarian torsion model.

OBJECTIVES: The aim of the study is to investigate the effects of Protocatechuic Acid (PCA), which is an antioxidant, anti-inflammatory and anti-apoptotic agent, on ovarian tissue and ovarian reserve against ischemia-reperfusion (IR) injury in a rat ovarian torsion model. BACKGROUND: Reactive oxygen radicals cause histopathological changes in the ovarian tissue during the reperfusion phase. PCA may have protective effects on ovarian tissue and reserve due to its antioxidant and antiapoptotic properties. METHODS: A total of 24 Wistar adult female rats were divided into 3 groups as the control (sham operation, n = 8), IR (Ischemia-Reperfusion, n = 8), and IR+PCA (Ischemia-Reperfusion + 80 mg/kg protocatechuic acid, n = 8). The IR and IR + PCA groups underwent 3 hours of ischemia followed by 3 hours of ovarian reperfusion. Protocatechuic acid (80 mg/kg) was administered to the IR+PCA group 30 minutes before reperfusion. After reperfusion, the ovaries were removed for histopathological and biochemical examination. RESULTS: Histopathological score and TUNEL+ cell count were significantly lower and AMH expression level was significantly higher in the IR+PCA group when compared to the IR group (p <0.05). However, in the comparison of the follicle counts, there was no statistically significant difference between all groups. Due to the increase in antioxidant activity, the MDA levels were found to be significantly lower in the IR+PCA group compared to the IR group (p < 0.05). CONCLUSION: Protocatechuic acid may be an effective antioxidant in protecting ovarian tissue and follicle reserve against IR injury of the ovary (Tab. 1, Fig. 4, Ref. 36).

Protective Effects of N-Acetyl Cysteine on Undescended Testis after Orchiopexy: A Rat-model Study.

ABSTRACT       Objective: To assess the effectiveness of utilising N-acetyl cysteine (NAC) to treat tissue damage brought on by undescended testis (UT) in rats after orchiopexy. STUDY DESIGN: Experimental study. Place and Duration of the Study: Bolu Abant Izzet Baysal University, Bolu, Turkey, from January 2018 to June 2020. METHODOLOGY: The UT model was created by administering flutamide to pregnant rats. Four groups of animals were created as the control group (offsprings of pregnant rats without flutamide), group II (UT), group III (UT + orchiopexy), and group IV (UT + orchiopexy + NAC); each containing eight animals. RESULTS: Group IV had a higher level of glutathione peroxidase than groups III and II (p=0.001 and p=0.002, respectively). Malondialdehyde was reduced in group IV compared with groups III and II (both p<0.001). There were differences in mean apoptotic cell counts (ACC) among the groups (p<0.001). ACC in group IV was lower than in group III (p<0.001). Sperm counts were higher in group IV than in groups III and II, and in group III they were higher than group II (p<0.001 all) and similar between groups IV and control group (p=0.102). CONCLUSION: Orchiopexy reduced UT-related testicular damage, additionally using NAC following orchiopexy may further reduce testicular damage through its antioxidant effects. KEY WORDS: Undescended testis, Testis damage, Orchiopexy, N-acetyl cysteine, Antioxidant.

Investigating the effects of carvacrol in rats using oxygen-induced retinopathy model.

Purpose: Investigating the effects of intraperitoneal carvacrol administration in rats using the oxygen-induced retinopathy (OIR) model. Methods: A total of 28 newborn Sprague Dawley rats were used and the OIR model was created using the 50/10% oxygen model. The study composed of four groups in total. While the OIR model was not used in Group I (control group), it was created for Groups II, III, and IV. About 0.01 mL carvacrol, bevacizumab, or 0.9% NaCl was administered intraperitoneal (IP) to the rats in all groups on postnatal day (PND) 14 as follows: Group I and Group II were administered 0.9% NaCl, Group III was administered bevacizumab, and Group IV was administered carvacrol. On PND 18, rats were sacrificed and their right eyes were enucleated. Results: Histopathological and immunohistochemical studies showed that the number of vascular endothelial cells (VECs), vascular endothelial growth factor (VEGF), and tumor necrosis factor-α (TNF-α) decreased similarly in Group III and Group IV compared with Group II. VECs values for Group I, Group II, Group III, and Group IV were measured as 0 ± 0, 26.45 ± 4.57, 7.75 ± 1.98, and 5.78 ± 1.72, respectively, and it differed significantly between groups (P < 0.001). Likewise, VEGF levels were observed as 0.06 ± 0.01, 3.31 ± 0.53, 2.47 ± 0.44, and 2.49 ± 0.52, respectively, and it differed significantly between groups (P < 0.001). TNF-α levels were recorded as 0.06 ± 0.01, 3.58 ± 0.38, 2.46 ± 0.49, and 2.29 ± 0.25, respectively, and it differed significantly between groups (P < 0.001). VECs, VEGF, and TNF-α were similar between Group III and IV (range of P values were 0.486-0.998). Conclusion: The study demonstrated that carvacrol significantly reduced retinal pathological angiogenesis, NV, VEC nuclei count, VEGF, and TNF-α levels. Moreover, the observed effects were comparable to those of bevacizumab.

Effects of isotretinoin on new bone formation after maxillary sutural expansion.

PURPOSE: The aim of this study was to examine the effects of isotretinoin on new bone formation after maxillary sutural expansion in rats. MATERIALS AND METHODS: A total of 32 male Wistar rats were selected and randomly divided into four groups. The isotretinoin group was treated with 7.5 mg/kg isotretinoin, and the soybean group was treated with 2 ml/kg soybean oil for 57 days. The substances were applied via oral gavage. The expansion-only and the control groups were not treated with any substance. In the experiment groups, expansion springs were applied on day 41 of the experiment, and after day 5 of expansion, a 12-day retention period was established. At the end of the experiment, all the animals were sacrificed, and their maxillae were dissected for histological evaluation. The numbers of osteoclasts, osteoblasts and formation of new bone and capillaries were evaluated on slides centered around the suture. RESULTS: The statistical analysis showed significant differences between the groups for the number of osteoblasts and osteoclasts (p < 0.001). In the experiment groups, higher numbers of osteoblasts and osteoclasts were detected in comparison to the control group (p < 0.001), while there was no significant difference between the experiment groups. Capillary formation and new bone formation in the isotretinoin group were found to be on a higher level than in the other groups (p < 0.001). CONCLUSION: Isotretinoin had no negative effects on bone formation following the expansion of the maxillary suture in rats.

High quality human sperm selection for IVF: A study on sperm chromatin condensation.

The study consisted of semen samples of 20 male individuals who applied to Abant Izzet Baysal University Faculty of Medicine and participated in a spermiogram. The aim of this study was to determine how to obtain the healthiest spermatozoa by employing a variety of swim-up methods over differing time periods and without the use of centrifuge. Ejaculate samples were taken from the 20 patients and each patient's homogenized semen sample was divided into 4 groups without centrifugation. Group 1 was taken as the sample of untreated semen. For the other 3 groups, 250 µl of medium was added in the semen samples. Afterwards, the samples were kept at 37 °C for different time periods, 30 min for Group 2, 60 min for Group 3 and 90 min for Group 4 in order for the spermatozoa to swim to the media in the upper layer. At the end of the periods, 10 µl of propagation preparations were prepared from the swim-up fluid. Using Aniline Blue for chromatin condensation analysis, two hundred cells were immunostained by Caspase 3 for apoptotic analysis. Subsequently, the result of the four groups were compared for each test. The spermatozoa obtained at the end of the 30 min. of swim-up was compared to the spermatozoa obtained from the swim-up of 60 min., the swim-up of 90 min. It was found that the control group had statistically significant lower rates of apoptosis and was healthier in terms of chromatin integrity. The swim-up method without centrifugation is the best suited sperm preparation, based on sperm DNA integrity and sperm chromatin condensation.

Major Ozonated Autohemotherapy Preconditioning Ameliorates Kidney Ischemia-Reperfusion Injury.

Medical ozone has therapeutic properties as an antimicrobial, anti-inflammatory, modulator of antioxidant defense system. Major ozonated autohemotherapy (MOA) is a new therapeutic approach that is widely used in the treatment of many diseases. The objective of the present study was to determine whether preischemic application of MOA would attenuate renal ischemia-reperfusion injury (IRI) in rabbits. Twenty-four male New Zealand white rabbits were divided into four groups, each including six animals: (1) Sham-operated group, (2) Ozone group (the MOA group without IRI), (3) IR group (60 min ischemia followed by 24 h reperfusion), and (4) IR + MOA group (MOA group). The effects of MOA were examined by use of hematologic and biochemical parameters consisting of neutrophil to lymphocyte ratio (NLR), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), ischemia-modified albumin (IMA), total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI). In addition, the histopathological changes including the tubular brush border loss (TBBL), tubular cast (TC), tubular necrosis (TN), intertubular hemorrhage and congestion (IHC), dilatation of bowman space (DBS), and interstitial inflammatory cells infiltration (IECI) were evaluated. In the IR group, compared to the Sham group, biochemical parameters indicating oxidative stress, NLR, IL-6, TNF-α, IMA, TOS, and OSI have increased. MOA reduced inflammation and oxidative stress parameters. Although TAS values have decreased in the IR group and increased in the MOA-pretreated group, no significant changes in TAS values were detected between the IR and MOA groups. The total score was obtained by summing all the scores from morphological kidney damage markers. The total score has increased with IR damage when compared with the Sham group (13.83 ± 4.30 vs 1.51 ± 1.71; p = 0.002). But, the total score has decreased significantly after application of MOA (5.01 ± 1.49; p = 0.002; compared with the IR group). MOA preconditioning is effective in reducing tissue damage induced in kidney ischemia-reperfusion injury. The protective effect of MOA is mediated via reducing inflammatory response and regulating of reactive oxygen species (ROS). Renal histology also showed convincing evidence regarding MOA's protective nature against kidney injury induced renal ischemia-reperfusion. Consequently, MOA might be helpful in protecting the kidneys from IR-induced damage in humans, probably through the anti-inflammatory effect and reducing the total oxidant status.