Investigation of the effect of rhamnetin on mice injected with solid and ehrlich ascites tumor.

Rhamnetin is a flavonoid which contained in especially clove, such as apple, tea, and onion plant. Rhamnetin has been used in cancer research due to its antitumor and antioxidant properties. In this study, effects of rhamnetin administration at different doses on ascites and solid tumors were investigated in Balb/C mice bearing EAT model that originating from rat breast adenocarcinoma. Experimental procedure: Overall, 92 Balb-c mice were used in this study. EAT cells (1 × 106 cells) that harvested from stock animals were injected to all rats via intraperitoneal and subcutaneous route. Rhamnetin (100 µg/kg-200 µg/kg) were given intraperitoneally and subcutaneously during 10 and 15 days to the animals bearing ascites tumor and solid tumor, respectively. Throughout experiments, weight changes were recorded in all groups. The maximum weight increase was observed in the control group among all groups (ascites and solid tumor groups). In the treatment groups, the least weight increase were determined in 200-µg/kg rhamnetin applied. The lowest increase in tumor volume was observed in the group that received 200-µg/kg rhamnetin (2.84) when compared to tumor control group (3.67). Result and conclusion: We determined that the number of live and dead cells in the treatment groups administered with the mean rhamnetin dose (2.5 µg/ml) was found in the count made in the EAT cell line after the incubation periods. We observed that rhamnetin plays an important role against cancer formation. We have obtained important results in our study, but detailed studies on the relationship between rhamnetin and cancer are needed.

Chondrotoxic effects of intra-articular injection of local anaesthetics in the rabbit temporomandibular joint.

The aim of this study was to investigate the chondrotoxic effects of a single-dose intra-articular injection of articaine, lidocaine, and bupivacaine on the rabbit temporomandibular joint (TMJ). Twenty-four rabbits were divided into four groups: control (group 1), articaine (group 2), lidocaine (group 3), and bupivacaine (group 4). Synovial fluid samples and venous blood were taken to evaluate matrix metalloproteinase 3 (MMP-3) levels. One millilitre of local anaesthetic solution was injected in the study groups and saline solution in the control group. The rabbits were euthanized after 4 weeks and the mandibular condyles and articular discs were evaluated. On histological examination, the study group samples had irregular joint surfaces, decreased collagen, and a thinner cartilage layer. Apoptotic cells were evaluated with the TUNEL method. TUNEL-positive apoptotic cell counts were higher in all study groups compared to the control group, and the difference was significant (P < 0.001). The mean preoperative serum MMP-3 level for all groups was 5.71 ± 3.33 ng/mL, while the mean postoperative level was 22.61 ± 6.36 ng/mL; this difference was significant (P < 0.001). A single-dose intra-articular injection of local anaesthetic had apoptotic effects on chondrocytes, leading to degenerative changes in the TMJ articular structures. Articaine was found to have less harmful effects than lidocaine and bupivacaine. Intra-articular injection of local anaesthetics should be limited in the TMJ because of the potential toxic effects.

The protective role of curcumin against toxic effect of nonylphenol on bone development.

INTRODUCTION: In the study, it was aimed to investigate the possible protective effects of curcumin, a potent antioxidant, against the toxic effect of nonylphenol on bone development. METHODS: Thirty pregnant female Wistar albino rats were used. The rats were randomly divided into the following five groups; the control group, corn oil group (150 µl/kg/day), nonylphenol group (50 µl/kg/day), curcumin group (100 mg/kg/day) and curcumin + nonylphenol group (100 mg/kg/day + 50 µl/kg/day). The doses were given by gavage from the 5th day to the 20th day of gestation. The fetuses were removed out on the 20th day of pregnancy by cesarean at the end of the study. After the sacrifice of the animals, double skeletal staining in front extremity (clavicula, scapula, humerus, radius, ulna) and hind extremity (femur, tibia, fibula), additionally histological and immunohistochemical examinations in femur bone were performed. RESULTS: The nonylphenol group offspring have the lowest weights of fetuses and placenta, head-to-hip lengths, biparietal and occipitofrontal length, and also, bone length percentage and percentage of the ossification area in all measurements of the front extremity and hind extremity Interestingly, the groups treated with curcumin showed close to the control group in terms of double skeletal staining, histological, and immunohistochemical examinations. CONCLUSIONS: Our findings demonstrated an association between bone development and exposure to nonylphenol. The findings suggest that curcumin treatments may be effective in accelerating bone formation.

The investigation of the antitumoral effect of Cornus mas L in mice with ehrlich solid tumor.

AIM: Cornus mas L is commonly used due to its anti-inflammatory, anti-carcinogenic and anti-oxidant properties. In the study, the effects of C. mas L extract on a solid tumor were examined in the Ehrlich solid tumor model developed in Balb/C type mice. METHODS: Ehrlich acid tumor (EAT) cells (1x106 EAT cell) from the stock animal were injected subcutaneously (s.c.) through the nape of the mice. Treatment groups of solid tumor-induced animals received 100 mg/kg and 200 mg/kg of C. mas L extract intraperitoneally (i.p.) for 14 days. RESULTS: Tumor volumes and animal weights were found to be statistically significant compared to the control group (p < 0.05). AgNOR staining was performed in tumor tissues. Statistically significant differences were observed between the groups in terms of TAA/NA ratio (p < 0.05). Immunohistochemical and biochemical parameters were also evaluated. An estimation of tumor proliferation of the lung, liver, brain, kidney, testis and tumor antioxidant parameters viz. lipid peroxidation, reduced glutathione (GSH), glutathione S-transferase (GST), superoxide dismutase (SOD) and catalase (CAT) was made. CONCLUSIONS: Our study showed that the anti-tumor effect of C. mas L in assisted tumor development with EAT cells, was mediated by the enhancement of oxidative stress with multiple mechanisms (Tab. 6, Fig. 12, Ref. 38).

The effects of N-acetylcysteine on cisplatin induced cardiotoxicity.

OBJECTIVES: Recent studies reported that oxidative stress is an important mechanism that contributes to cisplatin induced cardiotoxicity. In the present study, the effects of N-acetylcysteine (NAC), which is an antioxidant, on cisplatin induced cardiotoxicity were investigated in a rat model. METHODS: Thirty two rats were separated into 4 equal groups: Control, NAC-250, CP (cisplatin), CP+NAC. Rats in the experimental groups were treated with a single dose of cisplatin intraperitoneally (ip) (10 mg/kg) and NAC (ip, 250 mg/kg) for 3 consecutive days. At the end of the experiment, cardiotoxicity was determined from plasma CK-MB, LDH, cTnI and cardiac myosin light chain-1 (CMLC-1) levels. In the tissue samples, total oxidant capacity (TOC), total antioxidant capacity (TAC), lipid hydroperoxide (ROOH) and thiol levels were measured. The hearts were also analyzed histopathologically. RESULTS: It was determined that cisplatin increased the tissue TOC, ROOH levels and decreased TAC and thiol levels. NAC administration after cisplatin treatment was observed to have ameliorated histological and functional changes in heart. CONCLUSIONS: In conclusion, the results of this experimental study suggested that oxidative stress had a serious effect on cisplatin cardiotoxicity, and NAC could be used as a therapeutic agent in addition to standard cisplatin treatment protocols (Tab. 3, Fig. 1, Ref. 35).

Comparison of myotoxic effects of levobupivacaine, bupivacaine and ropivacaine: apoptotic activity and acute effect on pro-inflammatory cytokines.

We investigated the myotoxic effects of bupivacaine, ropivacaine and levobupivacaine on rat skeletal muscle and compared its apoptotic activity and acute effects on pro-nflammatory cytokines. We divided 40 Wistar albino rats into four equal groups. Rats were injected intramuscularly with 0.5% bupivacaine (group B), 0.5% ropivacaine (group R), 0.5% levobupivacaine (group L) or 0.9% normal saline (group SF). Animals were sacrificed on the second day after the injection. TNF-α, IL-1 and IL-6 levels were examined in muscle tissue using immunohistochemistry and immunofluorescence. Apoptotic cells were visualized by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. We found that levobupivacaine caused the lowest TNF-α, IL-1 and IL-6 expression levels, while the bupivacaine group caused the highest level compared to the other two agents. The greatest number of apoptotic cells was found in the bupivacaine group. Bupivacaine was more myotoxic than other anesthetic agents and increased apoptosis. The number of TUNEL positive apoptotic cells was lowest in the SF group. The greatest IL-1 immunoreactivity was found in the bupivacaine group. Bupivacaine and ropivacaine produced greater IL-6 expression than the SF group. Bupivacaine and ropivacaine caused greater TNF-α expression than the SF group, whereas the immunoreactivity of TNF-α was similar in the bupivacaine and ropivacaine groups.

Evaluation of therapeutic potential of intraperitoneal ozone gas in combination with insulin above cranial and spinal neuropathy in rats with diabetes mellitus.

OBJECTIVE: This study aimed to investigate the therapeutic effect of ozone in combination with insulin on cranial and spinal neuropathy in rats with diabetes mellitus (DM). MATERIALS AND METHODS: Sixty adult male Sprague Dawley rats were randomly divided into the following six groups (n = 10): control (C), ozone (O), diabetic (D), ozone-treated diabetic (DO), insulin-treated diabetic (DI), and ozone, insulin-treated diabetic (DOI). DM was induced by a single intraperitoneal (ip) streptozotocin injection (60 mg/kg), followed by 3 IU (ip) insulin administration for 60 days. Next, 1.1 mg/kg (50 µg/ml) ozone was administered to the O, DO, and DOI groups for 60 days. After inducing diabetes, the total oxidant status (TOS) and total antioxidant status (TAS) were measured; the oxidative stress index (OSI) was calculated. The brain and spinal cord tissues were obtained for histopathological evaluation. This cross sectional study was conducted in Dumlupinar University Laboratory Animals Research Center e.g 11.03.2015 ‒ 15.05.2015. RESULTS: TAS was higher in the DO, DI, and DOI groups than in the D group. TOS and OSI were lower in the DO, DI, and DOI groups than in the D group. Little pathological alterations with degenerated axons and vascular congestion were observed in the DO, DI, and DOI groups compared with the D group. CONCLUSION: Ozone with insulin can stimulate the endogenous antioxidant defense mechanism in diabetic neuropathy, thereby preventing reactive oxygen species-induced damage and protecting against cranial and spinal neuropathies (Fig. 6, Ref. 29).

Immunohistochemical investigation of P16, P53 and Ki-67's prognostic values in diffuse large B-Cell lymphomas.

AIM: The aim of this study is to determine the immunohistochemical properties of Ki-67, P53 expression and loss of P16, and to assess their relationship with both clinical parameters and patient survival in DLBCL. METHOD: Forty patients, diagnosed at the Pathology Department of our institute with nodal DLBCL were selected as the study group. The relationship between P16, P53, Ki-67 expressions and clinical and laboratory parameters like age, gender, performance status, Eastern Cooperative Oncology Group (ECOG), clinical stage, presence of B-symptoms, bone marrow involvement, International Prognostic Index (IPI) score, lactate dehydrogenase (LDH) level, extranodal extension, relapse, C-reactive protein (CRP), sedimentation, number of leukocytes in patients and patient survival were then statistically evaluated. RESULTS: Our results display no statistically significant correlation between P53 expression and loss of P16, Ki-67 proliferation index and clinical parameters and overall survival (p > 0.05). The only statistically significant relationship was between loss of P16 and stage (p 0.05). CONCLUSION: According to the results of our study, the loss of P16, P53 gene expression and Ki-67 proliferation index have no effect on life expectancy of patients with DLBCL (Tab. 2, Fig. 2, Ref. 29).

Assessment of femoral neck fractures in the elderly with respect to morphology and mineral density.

BACKGROUND: Femoral neck fractures are among the major orthopaedic problems seen in the elderly and the annual mortality rate is high. The calcium (Ca) and phosphorus (P) ratio can be used as an indicator of osteoporosis. The purpose of this study is to investigate the microarchitectural structure of the fractured regions of femoral head as well as bone mineral density in female and male patients. MATERIALS AND METHODS: The bone tissues taken from the fractured regions of 10 male and 9 female patients were examined with a scanning electron microscope. Electron probe microanalyses were carried out to measure mineral ratios. RESULTS: The bone trabeculae in the fractured area were thin and the cavities between trabeculae were seen to have transformed to irregular and broad structures. There were small valleculae reflecting osteoclastic activity. The analysis showed that the Ca/P ratio at the fracture site averaged 2.20/1 in women and 2.16/1 in men. As age increased, the percentage values of Ca and P decreased and the Ca/P ratio increased. CONCLUSIONS: Although there is no significant difference between the parameters of male and female patients, it seems that men can be affected by osteoporosis as much as women.

Effects of selenium on ischaemia-reperfusion injury in a rat testis model.

Selenium is shown to have beneficial effects on ischaemia-reperfusion (IR) injury. Our aim was to assess the effects of selenium on IR-induced testicular damage in terms of biochemical and histopathological evaluation. A total of 32 rats were randomised into four groups: control, IR, IR + selenium (IR + S) and S. Detorsion was applied after 3 h of torsion. Testicular tissue superoxide dismutase (SOD), glutathione (GSH), malondialdehyde (MDA), total antioxidant capacity (TAC) and DNA fragmentation levels were determined. Testicular tissue samples were examined by histopathological examination and terminal deoxynucleotidyl transferase dUTP nick end-labelling staining. The control, IR and IR + S groups had higher SOD values compared with the S group; SOD levels of the control and IR + S groups were higher than those of the IR group (P < 0.05). Further, MDA levels of the IR group were higher than those in the other three groups (P < 0.05). The IR group revealed lower TAC levels than the three groups (P < 0.05 for all). GSH levels of the IR group were significantly lower than those in the other three groups (P < 0.05 for all). In contrast, GSH levels of the IR + S group increased compared with those of the S group. The IR group had more DNA fragmentation than the control and S groups (P < 0.05). It is concluded that selenium possibly reduces oxidative stress and apoptosis caused by testicular IR injury in rats. The testicular protective effect of selenium appears to be mediated through its anti-apoptotic and antioxidative effects. However, selenium does not affect DNA fragmentation.

Effect of etodolac hydrazone, a new compound synthesised from etodolac, on spermatozoon quality, testicular lipid peroxidation, apoptosis and spermatozoon DNA integrity.

The aim of this study was to investigate the effect of etodolac hydrazone (EH), a new compound synthesised from etodolac, on spermatozoon quality, testicular lipid peroxidation, apoptosis and spermatozoon DNA integrity in rats. Group 1 (n = 8) received 1 ml dimethyl sulfoxide (DMSO) daily (Control); group 2 (n = 8) was treated with 5 mg kg(-1)  day(-1) EH, dissolved in 1 ml DMSO (EH-5); and group 3 (n = 8) was treated with 10 mg kg(-1)  day(-1) EH, dissolved in 1 ml DMSO (EH-10). All administrations were performed by gavage and maintained for 8 weeks. Both doses of EH administration caused significant decreases in absolute and relative weights of testis, whole epididymis, right cauda epididymis, and spermatozoon motility, spermatozoon count in comparison with the control group. Only 10 mg kg(-1)  day(-1) EH administration caused significant decreases in absolute and relative weights of seminal vesicles and serum testosterone level, and significant increases in testicular lipid peroxidation level, and numbers of TUNEL+ apoptotic germ cells and spermatozoa with damaged DNA along with some histopathological damages when compared to the control group. However, body and ventral prostate weight, and testicular antioxidant markers (glutathione, glutathione-peroxidase and catalase), were unaffected significantly by both doses of EH administration. In conclusion, two different doses of EH, in particular its high dose, damage to testicular spermatogenic cells and spermatozoon DNA and, it decreases spermatozoon motility, count and testosterone level in healthy rats.

The effects of adriamycin on E-cadherin mediated cell-cell adhesion and apoptosis during early kidney development.

Adriamycin (ADR) is strongly teratogenic. We investigated the effects of ADR on apoptosis and the intensity of E-cadherin expression in developing kidneys. An experimental group of rats was given 2 mg/kg/day ADR on days 6-9 of gestation and a control group was given saline on the same schedule. Embryos were decapitated on days 13, 15, 17 and 19 of gestation, and processed and embedded in paraffin for routine light microscopy. Kidney specimens were stained with hematoxylin and eosin or periodic acid-Schiff, or immunostained for E-cadherin. Apoptosis was assessed using the TUNEL method. Weight loss and developmental deficiency were determined in embryos of the experimental group. ADR damaged or destroyed tubule epithelial cells, which caused apparent dilatation of the tubule lumen. Also, the brush borders of proximal tubules were damaged and glomerular spaces were dilated. ADR caused apoptosis of kidney tissue by days 15, 17 and 19 of development and E-cadherin expression was up-regulated during kidney development compared to controls. We found that ADR can cause apoptosis and increased E-cadherin expression in the developing rat kidney. E-cadherin expression and apoptosis may contribute to the development of ADR nephrotoxicity.

Intermediate filament protein nestin is expressed in developing meninges.

BACKGROUND: Nestin is a type VI intermediate filament protein known as a marker for progenitor cells that can be mostly found in tissues during the embryonic and fetal periods. In our study, we aimed to determine the expression of nestin in meninges covering the brain tissue at different developmental stages and in the new born. METHODS: In this study 10 human fetuses in different development stages between developmental weeks 9-34 and a newborn brain tissue were used. Fetuses in paraffin section were stained with H+E and nestin immunohistochemical staining protocol was performed. RESULTS: In this study, in the human meninges intense nestin expression was detected as early as in the 9th week of development. Intensity of this expression gradually decreased in later stages of development and nestin expression still persisted in a small population of newborn meningeal cells. CONCLUSION: In the present study, nestin positive cells gradually diminished in the developing and maturing meninges during the fetal period. This probably depends on initiation of a decrease in nestin expression and replacement with other tissue-specific intermediate filaments while the differentiation process continues. These differences can make significant contributions to the investigation and diagnosis of various pathological disorders (Tab. 1, Fig. 3, Ref. 36).

Antioxidant effect of carnosine treatment on renal oxidative stress in streptozotocin-induced diabetic rats.

Nitric oxide (NO) plays a significant role in the development of diabetic nephropathy. We investigated the effects of an antioxidant, carnosine, on streptozotocin (STZ)-induced renal injury in diabetic rats. We used four groups of eight rats: group 1, control; group 2, carnosine treated; group 3, untreated diabetic; group 4, carnosine treated diabetic. Kidneys were removed and processed, and sections were stained with periodic acid-Schiff (PAS) and subjected to eNOS immunohistochemistry. Examination by light microscopy revealed degenerated glomeruli, thickened basement membrane and glycogen accumulation in the tubules of diabetic kidneys. Carnosine treatment prevented the renal morphological damage caused by diabetes. Moreover, administration of carnosine decreased somewhat the oxidative damage of diabetic nephropathy. Appropriate doses of carnosine might be a useful therapeutic option to reduce oxidative stress and associated renal injury in diabetes mellitus.