Distribution and morphologic characterization of telocytes in rat ovary and uterus: insights from ultrastructural and immunohistochemical analysis.

Telocytes (TCs) are characterized by a small oval-shaped cell body with long prolongations that are called telopods (Tps). PDGFR-ß and c-kit markers may assist for the immunohistochemical identification of TCs; however, by these means they cannot be identified with absolute specificity. Transmission electron microscopy (TEM) is considered as a gold standard method for TCs observation. Studies on TCs in the female reproductive system are limited, and there is a lack of awareness regarding TCs in rat ovaries. We aimed to demonstrate the existence and morphology of TCs in rat ovaries, alongside previously studied TCs in rat uteri. Thus, ovaries and uteri from young adult Sprague-Dawley female rats (n = 8) with regular estrous cycles were collected. Then, left ovaries and uteri were proccessed for TEM analysis, while the right ones were used for immunohistochemistry. As a result, TCs were seen throughout the rat's ovarian stroma with their characteristic cell bodies, Tps, podomes (Pds) and podomers (Pdms). Tps were situated within the thecal layer of the follicles, surrounding the corpus luteum and blood vessels. Ovarian TCs were recognized to have relationship with other TCs/stromal cells. Subsequently, TCs were seen in stroma of endometrium with surrounding blood vessels and uterine glands, myometrium and perimetrium in rat uteri. There was also no statistical significance between the number of c-kit+ and PDGFR-ß+ telocyte-like cells in both rat ovarian (p = 0.137) and endometrial stroma (p = 0.450). Further investigation of the roles and functions of TCs in the female reproductive system is needed.

Modified and alternative bone cements can improve the induced membrane: Critical size bone defect model in rat femur.

BACKGROUND: As a two-stage surgical procedure, Masquelet's technique has been used to care for critical-size bone defects (CSD). We aimed to determine the effects of modified and altered bone cement with biological or chemical enriching agents on the progression of Masquelet's induced membrane (IM) applied to a rat femur CSD model, and to compare the histopathological, biochemical, and immunohistochemical findings of these cements to enhance IM capacity. METHODS: Thirty-five male rats were included in five groups: plain polymethyl methacrylate (PMMA), estrogen-impregnated PMMA (E+PMMA), bone chip added PMMA (BC+PMMA), hydroxyapatite-coated PMMA (HA) and calcium phosphate cement (CPC). The levels of bone alkaline phosphatase (BALP), osteocalcin (OC), and tumor necrosis factor-alpha (TNF-α) were analyzed in intracardiac blood samples collected at the end of 4 weeks of the right femur CSD intervention. All IMs collected were fixed and prepared for histopathological scoring. The tissue levels of rat-specific Transforming Growth Factor-Beta (TGF-ß), Runt-related Transcription Factor 2 (Runx2), and Vascular Endothelial Growth Factor (VEGF) were analyzed immunohistochemically. RESULTS: Serum levels of BALP and OC were significantly higher in E+PMMA and BC+PMMA groups than those of other groups (P = 0.0061 and 0.0019, respectively). In contrast, TNF-α levels of all groups with alternative bone cement significantly decreased compared to bare PMMA (P = 0.0116). Histopathological scores of E+PMMA, BC+PMMA, and CPC groups were 6.86 ± 1.57, 4.71 ± 0.76, and 6.57 ± 1.51, respectively, which were considerably higher than those of PMMA and HA groups (3.14 ± 0.70 and 1.86 ± 0.69, respectively) (P < 0.0001). Significant increases in TGF-ß and VEGF expressions were observed in E+PMMA and CPC groups (P = 0.0001 and <0.0001, respectively) whereas Runx2 expression significantly increased only in the HA group compared to other groups (P < 0.0001). CONCLUSIONS: The modified PMMA with E and BC, and CPC as an alternative spacer resulted in a well-differentiated IM and increased IM progression by elevating BALP and OC levels in serum and by mediating expressions of TGF-ß and VEGF at the tissue level. Estrogen-supplemented cement spacer has yielded promising findings between modified and alternative bone cement.

The effects of the combination of temozolomide and Eribulin on T98G human glioblastoma cell line: an ultrastructural study.

Glioblastoma tumors are the most aggressive primary brain tumors that develop resistance to temozolomide (TMZ). Eribulin (ERB) exhibits a unique mechanism of action by inhibiting microtubule dynamics during the G2/M cell cycle phase. We utilized the T98G human glioma cell line to investigate the effects of ERB and TMZ, both individually and in combination. The experimental groups were established as follows: control, E5 (5 nM ERB), T0.75 (0.75 mM TMZ), T1 (1.0 mM TMZ), and combination groups (E5+T0.75 and E5+T1). All groups showed a significant decrease in cell proliferation. Apoptotic markers revealed a time-dependent increase in annexin-V expression, across all treatment groups at the 48-hour time point. Caspase-3, exhibited an increase in the combination treatment groups at the 48-hour mark. Transmission electron microscopy (TEM) revealed normal ultrastructural features in the glioma cells of the control group. However, treatments induced ultrastructural changes within the spheroid glioblastoma model, particularly in the combination groups. These changes included a dose-dependent increase in autophagic vacuoles and apoptotic morphology of the cells. In conclusion, the similarity in the mechanism of action between ERB and TMZ suggests the potential for synergistic effects when combined. Our results highlight that this combination induced severe damage and autophagy in glioma spheroids after 48 hours.

Effects of vitamin K2 administration on guided bone regeneration in diabetic rats.

AIM: The present study aimed to investigate the histomorphometric and immunohistochemical impacts of vitamin K2 on guided bone regeneration (GBR) in calvarial critical-size defects (CSDs) in diabetic rats. METHODS: A total of 30 rats were used in this study, comprising 12 non-diabetic (control) rats and 18 with streptozotocin-nicotinamide-induced experimental Diabetes mellitus (DM). In all rats, two calvarial CSDs were created: one defect was left empty (E), the other was treated with bovine-derived bone graft and collagen-based resorbable membrane (GM). Study groups were as follows: control rats administered saline (n = 6, C-E and C-GM groups) or vitamin K2 (n = 6, CK-E and CK-GM groups) and diabetic rats administered saline (n = 6, DM-E and DM-GM groups) or vitamin K2 (n = 6, DMK-E and DMK-GM groups). After 4 weeks of saline or vitamin K2 administration, the rats were euthanized. Bone defect healing and new bone formation were assessed histomorphometrically, and osteocalcin and osteopontin levels were examined immunohistochemically. RESULTS: Percentage of new bone formation was greater in CK-GM vs. CK-E and in DMK-GM vs. DMK-E [d = 3.86 (95% CI = 16.38-28.61), d = 1.86, (95% CI = 10.74-38.58), respectively, p < .05]. Bone defect healing scores were higher in CK-GM vs. CK-E and in DMK-GM vs. DMK-E [d = 2.69 (95% CI = -2.12 to -0.87), d = 3.28 (95% CI = 0.98-1.91), respectively, p < .05]. Osteocalcin expression levels were elevated in CK-GM vs. CK-E, in DMK-GM vs. DMK-E [d = 1.19 (95% CI = 0.08-1.41), d = 1.10 (95% CI = 0.02-1.22), respectively p < .05]. Vitamin K2 enhanced osteocalcin expression levels in DMK-E vs. DM-E [d = 2.78, (95% CI = 0.56-1.53), p < .05] and in DMK-GM vs. DM-GM [d = 2.43, (95% CI = 0.65-2.10), p < .05]. Osteopontin expression was enhanced in defects treated with GM vs. E defects [C-GM vs. C-E, d = 1.56 (95% CI = 0.38-2.01); CK-GM vs. CK-E, d = 1.91 (95% CI = 0.49-1.72); DM-GM vs. DM-E, d = 2.34 (95% CI = -1.12 to -0.50); DMK-GM vs. DMK-E, d = 2.00 (95% CI = 0.58-1.91), p < .05]. CONCLUSION: The research findings suggest that administering vitamin K2 in GBR for rats with DM favorably impacts bone healing in CSDs, presenting an adjunctive strategy for bone regeneration.

Evaluation of combined effects of brief electrical stimulation and Schwann-like cells on sciatic nerve injury model.

Severe nerve injuries can be treated with electrical stimulation and stem cell therapies, but little is known about the potential benefits of combining these two treatments. In an effort to investigate this combination, we conducted a study to evaluate the effectiveness of electrical stimulation and Schwann-like cell transplantation in female Wistar albino rats. Our study consisted of five groups of rats: a sham group, an injury group, an electrical stimulation group, a Schwann-like cell group, and a combination group. The experimental groups received electrical stimulation, Schwann-like cell transplantation, or both. The animals sciatic function index was evaluated during a 6-week recovery period, and nerve conduction velocity, wet muscle mass, and nerve tissues were also analyzed. The results of the study showed that all experimental groups had a faster functional recovery compared to the injury group, although the difference between groups was not statistically significant. Both the combination group and the Schwann-like cell transplantation group had a higher nerve conduction velocity compared to the other experimental groups. However, there was no significant difference between the combination and Schwann-like cell transplantation groups. Nonetheless, histological analysis showed a better axonal reorganization in the combination group. The study provides preliminary evidence of the potential benefits of combining electrical stimulation and Schwann-like cell transplantation in treating severe nerve injuries. However, further studies with larger sample sizes are needed to confirm these findings and optimize the treatment parameters.

Human ovarian granulosa cells use clathrin-mediated endocytosis for LDL uptake: immunocytochemical and electron microscopic study.

The steroidogenic activity of the granulosa cells is important for the reproductive cycle, and lipoproteins are involved in this process. The clathrin-mediated endocytosis pathway for LDL transport is considered to be the main one in eukaryotic cells. However, there are no studies that elucidate LDL internalization in human granulosa cells clarifying whether the clathrin-mediated endocytic pathway is functional in this process. The aim of this study is to investigate the role of clathrin and v-SNARE proteins in the formation of vesicles in human granulosa cells. In this study, the COV434 human granulosa cells were cultured and divided into four groups where in some of the groups Dil-conjugated LDL and Icarugamycin (ICA) a clathrin-mediated endocytosis inhibitor were added. From the collected mediums pregnenolone and progesterone levels were measured using ELISA. Oil red O staining was performed to show the intracellular lipids in the cells. Clathrin-coated vesicles believed to be responsible for carrying LDL, and v-SNARE proteins that direct the vesicles to their target molecules were also labeled and investigated by histological and ultrastructural methods. Our results show that human granulosa cells as well use the LDL cholesterol for steroid biosynthesis and they may prefer the clathrin-mediated endocytotic pathway to internalize it.

Investigation of apoptotic and antiproliferative effects of Turkish natural tetraploids Trifolium pratense L. extract on C6 glioblastoma cells via light and electron microscopy.

Glioblastoma (GBM) is the most common type of primary brain tumors in adults, characterized by its ability to proliferate rapidly and its tendency to aggressively and strongly invaded the surrounding brain tissue. The standard treatment approach of GBM is surgical resection followed by simultaneous chemotherapy and radiation. However, a significant number of GBM cases develop resistance to currently used chemotherapeutic drugs. Therefore, there is a need for the development of new chemotherapeutic agents. Trifoliumpratense L. is an endemic plant containing various isoflavones such as biochanin A, genistein, daidzein, and formononetin in high concentrations, and it has been shown in various studies that these molecules can function as anticancer agents. The present study was designed to determine the effect of the possible anticarcinogenic effects of the Trifolium pratense L. which grown in our country and to obtain new treatment approaches alternative to the classical treatment protocols applied in the treatment of GBM. C6 glioblastoma cells were cultured with Trifolium pratense L. Cell proliferation, apoptotic cell morphology, and cell structure were evaluated with CCK8, Annexin V, cytochrome c, CD117, and Betatubulin labeling, respectively. And also, investigated effects of this Turkish tetraploid on GBM by TEM. Decreased cell proliferation and increased number of apoptotic cells were observed depending on the increasing doses of Trifolium pratense L. In addition, intense morphological changes were detected depending on increasing doses. In this context, we believe that the plant Trifolium pratense L., may be a new alternative and adjuvant agent for the treatment of GBM.

Effects of subepineural hyaluronic acid injection on nerve recovery in a rat sciatic nerve defect model.

BACKGROUND: Maintenance of epineural integrity is very important for nerve healing. Reports on the use of substances consid-ered to have positive effects on nerve healing in experimental nerve defect models are increasing. The present study assessed the effects of sub-epineural hyaluronic acid injection in a rat sciatic nerve defect model that was created while maintaining epineural integrity. METHODS: The study included 40 Sprague Dawley rats. The rats were randomly divided into a control group and three experimental groups (10 rats in each group). In the control group, the sciatic nerve was dissected and no additional surgery was performed. In experimental group 1, the sciatic nerve was transected in the middle, and then, primary repair was performed. In experimental group 2, a 1-cm defect was created while preserving the epineurium, and then, the defect was repaired with end-to-end suturing of the pre-served epineurium. In experimental group 3, the surgical procedure for experimental group 2 was performed, and then, sub-epineural hyaluronic acid injection was carried out. Functional and histological evaluations were performed. RESULTS: On functional evaluation, there was no statistically significant difference among the groups during the 12-week follow-up period. On histological evaluation, nerve recovery was poorer in experimental group 2 than in experimental groups 1 and 3 (p<0.05). CONCLUSION: Although the functional analysis did not reveal any significant results, the histological findings suggest that hyaluronic acid increases the regeneration capacity of axons through its anti-fibrotic and anti-inflammatory effects.

The antibiotics supplemented bone cement improved the masquelet's induced membrane in a rat femur critical size defect model.

BACKGROUND: Masquelet technique is a two-stage surgical procedure used in the treatment of critical-size bone defects (CSD). Adding antibiotics to polymethylmethacrylate (PMMA) is still questionable to create higher quality induced membrane (IM). The aim of the study was to evaluate the effects of three antibiotic-supplemented cement, fusidic acid, teicoplanin, and gentamicin, on osteogenesis and IM progression applied to rat femur CSD model by comparing histopathological, biochemical, and immunohistochemical findings. METHODS: Twenty-eight male rats were divided into four groups control, gentamicin (G), teicoplanin (T), and fusidic acid (FA). A 10 mm CSD was created in rat femurs. In the postoperative 4th week, intracardiac blood samples were collected for biochemical analysis of bone alkaline phosphatase (BALP), osteocalcin (OC), and tumor necrosis factor-alpha (TNF-α) levels. IMs obtained in secondary operation were fixed and prepared for histopathological scoring of membrane progression and immunohistochemical evaluation of rat-specific Transforming Growth Factor-Beta (TGF-ß), Runt-related Transcription Factor 2 (Runx2), and Vascular Endothelial Growth Factor (VEGF) expressions. RESULTS: Levels of BALP and OC in serum didn't change among groups significantly while serum TNF-α levels significantly decreased in all antibiotic groups compared to the control group (P = 0.017). Histological scores of groups FA and T were significantly higher than those of groups Control and G (P = 0.0007). IMs of groups T and FA showed good progression while those of groups Control and G were also moderately progressed. A significant increase in TGF-ß expression was observed in group G and FA (P = 0.001) while a significant increase in the expression of VEGF was observed in groups G and T compared to the control group (P = 0.036). CONCLUSIONS: The bone cement impregnated with thermostable and safe antibiotics, gentamicin, fusidic acid, and teicoplanin can increase osteogenesis and support IM progression by increasing the expressions of TGF-ß and VEGF. Anabolic effects of induced membranes used in the treatment of critical-size bone defects can be enhanced by antibiotic-supplemented PMMAs applied by altering the original technique.

Partial load-bearing rabbit ulnar segmental defects are regenerated with biocompatible grafts with or without bone marrow-derived mesenchymal stem cells.

BACKGROUND: The autologous bone grafts still have been used as the gold standard to initiate and facilitate bone healing in cases with bone defects. Because of some disadvantages of autologous bone grafts, the new biocomposite grafts have been researched. The purpose of the present study was to investigate whether the bone marrow-derived mesenchymal stem cells (BM-MSCs) loaded into a biocomposite scaffold enhance bone regeneration. METHODS: In our study, a 10 mm osteoperiosteal segmental bone defect was created in the middle of the right and left ulnar bones of eight rabbits. The created defects were filled in the right ulnar bones of eight rabbits (Group I) with BM-MSCs loaded onto a bio-composite scaffold (Plexur PTM, Osteotech, Eatontown, NJ, USA) and in the other ulnar bones of the same rabbits (Group II) with only biocomposite graft. Radiographs of each forelimb were taken postoperatively at the end of the 6th week. Then, the rabbits were euthanized pharmacologically for histopathological evaluation. RESULTS: Were scored using a modified Lane and Sandhu scoring system. All defects healed in both groups. Radiological and histo-logical total scores were slightly better in Group I, but statistical tests did not reveal any significant differences between the two groups at the end of the 6th week radiologically and histologically (p>0.05). CONCLUSION: The results of our study demonstrated that in rabbit ulnar segmental bone defect model was obtained satisfactory regeneration with using biocomposite graft with or without BM-MSCs.

Fresh Human Myopic Lenticule Intrastromal Implantation for Keratoconus Using SMILE Surgery in a Long-term Follow-up Study: Ultrastructural Analysis by Transmission Electron Microscopy.

PURPOSE: To investigate new intrastromal histological structures that develop after myopic human lenticular implantation in keratoconus with femtosecond laser-assisted small incision lenticule extraction (SMILE) surgery using transmission electron microscopy. METHODS: Sixty eyes with advanced keratoconus indicated for corneal transplantation were included in this study. Fresh myopic lenticular implants were placed in all eyes through SMILE surgery. Lenticular implants were extracted from patients with myopic refractive errors of the cornea, untreated keratoconus, and treated keratoconus following 1, 2, and 3 years of surgery. These five lenticular samples were examined under the electron microscope and compared. RESULTS: Disorganized and thinned collagen fibers were observed in the stroma with degenerative stromal cells (telocyte-like cells and keratocytes) in the keratoconic cornea. Apoptotic bodies and cell debris were easily observed near the disorganized fibers. In contrast, the myopic refractive error of the control and treatment groups demonstrated well-organized parallel lamellar structures. Healthy keratocytes and telocyte-like cells were observed in samples obtained 1, 2, and 3 years after lenticular implantation. Thus, telocyte-like cells may be activated by appropriate stimuli, such as stem cells, and be involved in stromal regeneration. CONCLUSIONS: Fresh myopic intrastromal lenticular implantation is a safe, economical, and reliable technique that leads to increased corneal thickness, improved visual acuity, and the regeneration of healthy keratocytes and telocyte-like cells that are involved in stromal regeneration. [J Refract Surg. 2022;38(8):520-528.].

Combined resveratrol and vitamin D treatment ameliorate inflammation-related liver fibrosis, ER stress, and apoptosis in a high-fructose diet/streptozotocin-induced T2DM model.

A high fructose diet is a major cause of diabetes and various metabolic disorders, including fatty liver. In this study, we investigated the effects of resveratrol and vitamin D (VitD) treatments on endoplasmic reticulum (ER) stress, oxidative stress, inflammation, apoptosis, and liver regeneration in a rat model of type 2 diabetes mellitus, namely, T2DM Sprague-Dawley rats. This T2DM rat model was created through a combination treatment of a 10% fructose diet and 40 mg/kg streptozotocin (STZ). Resveratrol (1 mg/kg/day) and VitD (170/IU/week) were administered alone and in combination to both the diabetic and control groups. Immunohistochemical staining was performed to evaluate PCNA, NF-κB, TNF-α, IL-6, IL-1ß, GRP78, and active caspase-3 in liver tissue. The TUNEL method and Sirius red staining were used to determine apoptosis and fibrosis, respectively. G6PD, 6-PGD, GR, and GST activities were measured to determine oxidative stress status. We found that the expressions of cytokines (TNF-α, IL-6, and IL-1ß) correlated with NF-κB activation and were significantly increased in the T2DM rats. Increased GRP78 expression, indicating ER stress, increased in apoptotic cells, enhanced caspase-3 activation, and collagen accumulation surrounding the central vein were observed in the T2DM group compared with the other groups. The combination VitD + resveratrol treatment improved antioxidant defense via increasing G6PD, 6-PGD, GR, and GST activities compared to the diabetic groups. We concluded that the combined administration of resveratrol with VitD ameliorates the adverse effects of T2DM by regulating blood glucose levels, increasing antioxidant defense mechanisms, controlling ER stress, enhancing tissue regeneration, improving inflammation, and reducing apoptosis in liver cells. In conclusion, this study indicates that the combination treatment of resveratrol + VitD can be a beneficial option for preventing liver damage in fructose-induced T2DM.

The value of potassium, pH and D-dimer levels in early diagnosis of acute mesenteric ischemia: an experimental study on rats.

INTRODUCTION: The aim of this randomized controlled experimental study was to evaluate the efficacy of potassium, pH and D-dimer levels in blood, as well as potassium and pH levels in peritoneal lavage fluid, in the early diagnosis of acute mesenteric ischemia. MATERIAL AND METHODS: This study was conducted at the Istanbul University Center of Experimental Medicine after having received approval from the Istanbul University animal testing ethics committee. Male albino Wistar rats (n = 24; 250 to 350 g) were divided into two control groups and two ischemic groups. Levels of potassium, pH, and D-dimer in blood and levels of potassium and pH in peritoneal lavage fluid were analyzed for 1 h and 2 h after the induced acute mesenteric ischemia procedure. The degree of ischemic injury was determined using the histopathological damage score in tissue samples taken from the terminal ileum. RESULTS: Ischemic groups had statistically significant differences in potassium and pH in blood and peritoneal lavage fluid compared to non-ischemic groups (p < 0.05). There was no significant difference between control and ischemic groups in terms of D-dimer and histologic grading results after 1 h (p = 0.132, p = 0.475 respectively), while there was a significant difference between control and ischemic groups after 2 h (p < 0.05). CONCLUSIONS: The levels of potassium, pH, and D-dimer could be useful in daily practice for the early diagnosis of acute mesenteric ischemia.

The Protective Effects of Thymosin-ß-4 in a Rat Model of Ischemic Acute Kidney Injury.

BACKGROUND: Despite the progress in the treatment of acute kidney injury (AKI), current curative approaches fail to provide adequate treatment. In this study, we aimed to investigate the possible protective effects of thymosin-ß-4(Tß4) on an ischemic AKI model in rats. METHODS: Rats were randomly assigned into four groups (n = 8/group): The control group (sham-operated), the ischemia-reperfusion (I/R) group; renal ischemia (90 min) by infrarenal abdominal aortic occlusion followed by reperfusion (3 h), the Tß4 + I/R group; treated with Tß4 before I/R, and the I/Tß4/R group; treated with Tß4 just before reperfusion. Besides renal function determination (creatinine (Cr) and blood urea nitrogen (BUN)); histological evaluation was also conducted. Renal tissue caspase-9, matrix metalloproteinase (MMP-9) activities, and hyaluronan levels were measured. Additionally, renal tissue oxidative stress (lipid hydroperoxide, malondialdehyde, superoxide dismutase, glutathione, pro-oxidant-antioxidant balance, ferric reducing antioxidant power, nitric oxide), inflammation (tumor necrosis factor-α, interleukin-1ß, interleukin-6, nuclear factor-κß) were evaluated. RESULTS: I/R increased the level of caspase-9, MMP-9 activity, and hyaluronan (p < 0.001) and these were significantly decreased in both Tß4 groups. Moreover, I/R led to increases in oxidative stress and inflammation parameters (p < 0.001) while the levels of antioxidants were decreased. Nevertheless, Tß4 in both groups were able to restore oxidative stress and inflammation parameters. Furthermore, Tß4 attenuated histologic injury caused by I/R (p < 0.01) and diminished serum urea-creatinine levels (p < 0.001). CONCLUSION: These results suggest that Tß4 has significant improving effects in ischemic acute kidney injury. This beneficial effect might be a result of the inhibition of extracellular matrix remodeling and apoptosis cascade via modulation in renal redox status and inflammation.

Protective effect of hyperbaric oxygen treatment on rat intestinal mucosa after mesenteric ischaemia and reperfusion.

INTRODUCTION: Mesenteric ischaemia results from a lack of adequate blood flow to and oxygenation of the mesentery and intestines. The aim of the present study was to evaluate the effect of hyperbaric oxygen treatment (HBOT) on the healing process in intestinal mucosa of rats undergoing mesenteric ischaemia and reperfusion. METHODS: Thirty-two Wistar-Albino rats were divided into four groups of eight: 1) ischaemia/reperfusion (I/R); 2) sham operation; 3) I/R+HBOT started 6 hours after reperfusion; 4) I/R+HBOT started 12 hours after reperfusion. In the I/R groups, a vascular clamp was placed across the superior mesenteric artery to occlude arterial circulation for 60 minutes, followed by reperfusion. A dose of HBOT consisted of 100% oxygen breathing for 90 minutes at 2.5 atmospheres absolute pressure. Thirteen doses of HBOT were administered after ischaemia. The rats were sacrificed on the eighth day, and their intestinal tissues were harvested for histopathologic analysis. The tissue levels of catalase, malondialdehyde, and glutathione were determined. RESULTS: The histopathological scores (HSCORE) were consistent with macroscopic examinations. The scores were significantly higher (worse) in Group 1 compared to Group 2, Group 3, and Group 4 (for all comparisons, P < 0.05). Group 4's HSCORE was significantly higher than those of Group 2 and Group 3 (for both comparisons P < 0.05). Group 3's HSCOREs were only marginally higher than Group 2. Group 3 exhibited higher glutathione levels than Group 1 (P < 0.05). There were no significant differences across the groups with respect to malondialdehyde and catalase levels. CONCLUSION: A beneficial effect of HBOT was observed on oxidative stress and inflammation in acute mesenteric ischaemia-reperfusion.

Protective Effect of Thymosin ß4 against Abdominal Aortic Ischemia-Reperfusion-Induced Acute Lung Injury in Rats.

Background and objectives: Ischemia-reperfusion (IR) caused by infrarenal abdominal aorta cross-clamping is an important factor in the development of ischemia-reperfusion injury in various distant organs. Materials and Methods: We investigated potential antioxidant/anti-inflammatory effects of thymosin beta 4 (Tß4) in a rat model of abdominal aortic surgery-induced IR. Tß4 (10 mg/kg, intravenous (i.v.)) was administered to rats with IR (90-min ischemia, 180-min reperfusion) at two different periods. One group received Tß4 1 h before ischemia, and the other received 15 min before the reperfusion period. Results: Results were compared to control and non-Tß4-treated rats with IR. Serum, bronchoalveolar lavage fluid and lung tissue levels of oxidant parameters were higher, while antioxidant levels were lower in the IR group compared to control. IR also increased inflammatory cytokine levels. Tß4 reverted these parameters in both Tß4-treated groups compared to the untreated IR group. Conclusions: Since there is no statistical difference between the prescribed results of both Tß4-treated groups, our study demonstrates that Tß4 reduced lung oxidative stress and inflammation following IR and prevented lung tissue injury regardless of timing of administration.

Protective effects of pentoxifylline in small intestine after ischemia-reperfusion.

Objective This study was performed to determine the healing effects of pentoxifylline on molecular responses and protection against severe ischemic damage in the small intestine. Methods Thirty-six Wistar albino rats were divided into six groups. The superior mesenteric artery was clamped for 120 minutes, and reperfusion was performed for 60 minutes. Saline (0.4 mL), pentoxifylline (1 mg/kg), and pentoxifylline (10 mg/kg) were intraperitoneally administered to the rats in the C1, P1, and P3 groups, respectively, 60 minutes before ischemia and to the rats in the C2, P2, and P4 groups, respectively, during reperfusion onset. Malondialdehyde, myeloperoxidase, tumor necrosis factor alpha, interleukin-1 beta, and interleukin-6 in serum and tissue were measured by enzyme-linked immunosorbent assay. Intestinal ischemic injury was histopathologically evaluated by the Chiu score and immunohistochemical staining. Results All serum and tissue molecular responses were significantly blunted in the pentoxifylline-treated groups compared with the controls. Significant improvement in ischemic damage was demonstrated in the pentoxifylline-treated groups by histological grading and immunohistochemical scoring. Conclusions The protective effects of pentoxifylline were confirmed by molecular responses and histopathological examination.

Effects of different drug treatments on the proliferation of human ovarian carcinoma cell line MDAH-2774

Background/aim: In this study, the effects of resveratrol as a natural polyphenol compound, gemcitabine as an antimetabolite that has nucleoside structure analogous to deoxycytidine, and para-aminophenol-derived paracetamol were investigated with single and combined applications in monolayers of the MDAH-2774 human ovarian cancer cell line. Materials and methods: Drugs were evaluated in cell culture with respect to cell proliferation, cell cytotoxicity (trypan blue dye exclusion test), synthesis phase of cell cycle, and cell structure in 24, 48, 72, and 96 h. Result: Resveratrol and gemcitabine diminished both cell proliferation and cell cycle synthesis phase indication in monolayer cell cultures (P < 0.05). All combination groups showed similar effects that were mainly more effective in respect to single usage of resveratrol and gemcitabine in monolayer cell cultures. Conclusion: The effects of gemcitabine, resveratrol, and paracetamol were investigated in monolayers of the MDAH-2774 human ovarian cancer cell line and a decrease in cell number in cell cycle synthesis phase, prevention of cell proliferation, and destruction of cell structure were observed.

Antioxidant activity of CAPE (caffeic acid phenethyl ester) in vitro can protect human sperm deoxyribonucleic acid from oxidative damage.

PURPOSE: Sperm processing (e.g., centrifugation) used in preparation for assisted reproduction can result in excessive generation of reactive oxygen species (ROS) and potential sperm damage. The use of antioxidants during sperm processing has been shown to prevent iatrogenic sperm damage, including DNA damage. In this study, we evaluated the effect of caffeic acid phenethyl ester (CAPE) on oxidative stress mediated sperm dysfunction and DNA damage. METHODS: Semen samples were obtained to liquefy at room temperature. After centrifugation and washing protocols, spermatozoa were incubated in a single step supplemented medium with either of 10, 50 or 100 µmol/L CAPE for 2 hours at 36 °C. After incubation period, MDA levels of seminal plasma were measured. The fragmentation in sperm DNA was detected by light microscopy via use of an aniline blue assay, while ultrastructural morphology was analyzed by transmission electron microscopy. RESULTS: Significant increase has been observed in percent chromatin condensation (assessed by aniline blue staining) and Malondialdehyde (Mmol/L) in oligoasthenoteratozoospermia group before the centrifugation (0.57 ±â€¯0.15). Incubation of samples with 100 µmol/L CAPE after centrifugation resulted in a significantly lower percent chromatin condensation compared to samples incubated without CAPE (0.42 ±â€¯0.12) (P < 0.0033). Incubation of all samples with CAPE (10 µmol/L, 50 µmol/L, 100 µmol/L.) after centrifugation resulted in a significantly lower percentage of Malondialdehyde levels. CONCLUSIONS: The data suggests that preincubation of spermatozoa with the antioxidant CAPE offers protection against oxidative DNA damage in vitro.

Effects of resveratrol on high-fructose-induced testis injury in rats.

This study investigated whether a high-fructose (HFr) diet changes the morphology of seminiferous tubules (ST) in rats and resveratrol (RES) has a possible restoring effect in this sense. Fructose (30%; w/v) was administered to rats alone or together with RES (50 mg/L) in drinking water for 8 weeks. In the HFr group, destruction of the germinal epithelium led to the detection of immature germ cells in the lumen. HFr diet gave rise to a decrease in the ST diameters (p < 0.05), Johnsen's tubular biopsy score values (p < 0.001), and an increase in the apoptotic index (p < 0.05). Ultrastructurally, HFr feeding increased lipid accumulation (p < 0.01), mitochondrial damage, and acrosomal abnormalities in spermatogenic cells. Treatment of HFr -fed rats with RES improved the reduced ST diameters and overall general histological and ultrastructural abnormalities of the STs, but did not change the increased apoptotic index.