Stereological Study on the Effect of Carnosine on of Purkinje Cells in the Cerebellum of Rats Exposed to 900 MHz Electromagnetic Field.

AIM: To evaluate the impact of carnosine on Purkinje neurons in rats exposed to a 900 Mhz electromagnetic field. MATERIAL AND METHODS: This study evaluated 24 rats divided into the following three different groups: a control group, a group exposed to the electromagnetic field, and a group that was injected with carnosine while being exposed to the electromagnetic field. The electromagnetic field group was exposed to a 900 Mhz electromagnetic field for an hour daily over 28 days. Thereafter, stereological analysis was performed histologically on cerebellar sections, and the number of Purkinje cells were counted. RESULTS: The electromagnetic field group had remarkably fewer Purkinje cell compared to control. The electromagnetic field group plus 20 mg of carnosine had significantly more total Purkinje cells compared to the electromagnetic field group (p < 0.05). CONCLUSION: The present study showed that electromagnetic field exposure decreases the number of Purkinje cell, whereas carnosine protected the cerebellum from neural damage induced by electromagnetic field exposure.

Effects of diabetes on nitric oxide synthase in rat uterus.

Diabetes mellitus (DM) is a common metabolic disorder with complications including nephropathy, cardiomyopathy, neuropathy and infertility. We investigated nitric oxide synthase (NOS) expression in uterine tissue of diabetic rats. The rats were assigned randomly to four groups of ten: group C, control; group 1, diabetic for 1 month; group 2, diabetic for 2 months; group 3, diabetic for 3 months. DM was induced by streptozotocin injection. nNOS, iNOS, eNOS expressions and TUNEL staining were assessed in uterine tissue and luteinizing hormone (LH), follicle stimulating hormone (FSH) and estrogen were measured in blood samples. The uterine epithelium, connective tissue of the endometrium, myometrium wall thickness and number of apoptotic cells were decreased in groups 1 - 3 compared to group C. nNOS and eNOS expression was increased in diabetic groups, but iNOS expression was similar to group 1. FSH was increased in group 2, but serum LH and estrogen levels were unchanged among groups. DM damages uterine tissue, and NOS, especially nNOS and eNOS, contributes to this damage.

The protective effects of thymoquinone on lung damage caused by cigarette smoke.

Chronic obstructive pulmonary disease (COPD) is characterized by systemic inflammation that usually is caused by exposure to noxious particles or gases. Thymoquinone (TQ) prevents the production of inflammatory mediators, such as thromboxane B2 and leukotriene, by altering arachidonic acid metabolism. We investigated the preventive and curative effects of TQ on lung damage in rats caused by cigarette smoke (CS). We used 50 adult male rats, 30 of which were exposed to CS every day for 3 months. TQ in dimethylsulfoxide (DMSO) was administered intraperitoneally (i.p.) every day to ten animals to investigate the protective effects of TQ, and to ten other animals during the last 21 days to investigate the curative effect. Ten rats received saline for the last 21 days. Ten subjects were untreated controls. Ten controls that were not exposed to CS received TQ for the last ten days. Serum IL-8, IL-6, IL-1ß and MMP-9 levels were measured using ELISA. IL-1ß and IL-8 levels were elevated in the group exposed to CS compared to controls. IL-8 levels were decreased in the group that received only TQ compared to controls, which indicated the anti-inflammatory effect of TQ. The apoptotic index (AI) was increased in all groups that were exposed to CS compared to controls. The AI index was decreased in the group that received TQ for the last 21 days compared to the other CS groups. AI was increased in the group that received TQ daily compared to the other CS groups. Our findings indicate that TQ exerts curative effects for the inflammation caused by CS and may prevent apoptosis if administered in appropriate doses; however, long term TQ or DMSO exposure may produce cumulative toxic effects.

Assessment of the Effectiveness of Quercetin on Cisplatin-Induced Ototoxicity in Rats.

OBJECTIVES: This study aimed to evaluate the effect of quercetin on cochlear function and morphology, and its possible protective effect against acute cisplatin-induced ototoxicity in rats. MATERIALS AND METHODS: This prospective and controlled animal study was conducted on Wistar albino rats divided into four groups. Otoacoustic emission measures were performed three days after the first infiltration in Group 1 (saline), 2 (cisplatin), and 3 (quercetin). This interval was five days for Group 4 (cisplatin+quercetin). At the end of the study, the rats were decapitated with deep anesthesia, and histological changes in the cochleas were observed by light microscopy. RESULTS: Group 2 (cisplatin) revealed significant differences between the first and second measures in all frequencies. When compared to other group, the difference of the changes in Group 2 statistically significantly decreased, especially in higher frequencies. Morphologically, there were no acute changes in Group 1 and Group 3. Outer hair cell loss and the degeneration of stria vascularis and spiral ganglion were observed in both Groups 2 and 4; the damages in the latter were lesser. CONCLUSION: Quercetin does not have negative effect on cochlea, and it has protective effect on cisplatin-induced ototoxicity.

Protective effect of vitamin D against hyperoxia-induced lung injury in newborn rats.

INTRODUCTION: Preterm infants have risks of developing vitamin D deficiency. Thus we aimed to investigate the effect of vitamin D on hyperoxia-induced lung injury in newborn rats. METHODS: Full term rat pups were included in the study 12-24 hr after delivery. The pups were randomly divided into eight groups as follows: normoxia control group (NC), normoxia plus vitamin D group (ND1, 1 ng/gr/day vitamin D), normoxia plus vitamin D group (ND2, 3 ng/gr/day vitamin D), normoxia plus vitamin D group (ND3, 5 ng/gr/day vitamin D), hyperoxia control group (HC), hyperoxia plus vitamin D group (HD1, 1 ng/gr/day vitamin D), hyperoxia plus Vitamin D group (HD2, 3 ng/gr/day vitamin D), hyperoxia plus vitamin D group (HD3, 5 ng/gr/day vitamin D). The histopathological effects of vitamin D were assessed by alveolar surface area (with mean linear intercept (MLI) method), apoptosis index and proliferating cell nuclear antigen (PCNA) index. RESULTS: MLI values were significantly lower among three groups (HD1: 83.93 ± 1.95 µm, HD2: 81.76 ± 1.68 µm, and HD3: 82.33 ± 1.87 µm) when compared with HC group (92.98 ± 2.09 µm) (P = 0.001, P = 0.0004, P = 0.002, respectively). Apoptotic cell index were significantly lower among three treatment groups (HD1: 1.455 ± 0.153, HD2: 0.575 ± 0.079, and HD3: 0.700 ± 0.105) when compared with HC group (2.500 ± 0.263) (P = 0.001, P = 0.001, P = 0.001, respectively). Although PCNA positive cell index did not change in HD1 group (0.132 ± 0.008) (P > 0.05), there were significant increases in HD2 (0.277 ± 0.026) and HD3 (0.266 ± 0.018) group when compared with HC group (0.142 ± 0.010) (HD2 P = 0.001, HD3 P = 0.001). CONCLUSION: Vitamin D seems to protect hyperoxia-induced lung injury in newborn rats. Pediatr Pulmonol. 2017;52:69-76. © 2016 Wiley Periodicals, Inc.

Long Term Exposure to Myrtucommulone-A Changes CD105 Expression and Differentiation Potential of Mesenchymal Stem Cells.

Mesenchymal stem cells (MSCs) represent a heterogeneous group of multipotent stem cells that could be found in various somatic tissues. MSCs are defined by molecular and functional features including spindle-shape morphology, adherence to plastic surfaces, expression of specific surface markers and differentiation potential to chondrocytes, adipocytes and osteocytes. The surface markers were proposed to affect the differentiation potential of MSCs by a limited number of studies. Endoglin (CD105) is defined to be a significant marker for osteogenic and chondrogenic differentiation ability of MSCs. Low CD105 expression is associated with increased osteogenic potential while high CD105 expression is correlated with strong chondrogenic potential. Myrtucommulone-A (MC-A) is an active compound with various biological effects on different cell types but its effect on MSC differentiation has not been described yet. In the present study we aimed at investigating the long-term effects of MC-A on hMSCs. MC-A-treatment reduced CD105 expression in distinct human mesenchymal stem cell (hMSC) lines and gave rise to CD105low population but did not change CD44, CD90 or CD73 expression. The decrease in CD105 expression reduced the chondrogenic potential of hMSCs subsequently while adipogenic or osteogenic differentiation was not affected dramatically. MC-A-treatment also suppressed the NF-κB p65 activation which might be responsible for the reduced chondrogenic potential. Our findings suggest that MC-A could be used to enrich CD105low hMSCs without the need for cell sorting or changing culture conditions which could be utilised in targeted differentiation studies.

Effects of cell-mediated osteoprotegerin gene transfer and mesenchymal stem cell applications on orthodontically induced root resorption of rat teeth.

Aim: The aim of this study is to evaluate and compare therapeutic effects of mesenchymal stem cell (MSCs) and osteoprotegerin (OPG) gene transfer applications on inhibition and/or repair of orthodontically induced inflammatory root resorption (OIIRR). Materials and methods: Thirty Wistar rats were divided into four groups as untreated group (negative control), treated with orthodontic appliance group (positive control), MSCs injection group, and OPG transfected MSCs [gene therapy (GT) group]. About 100g of orthodontic force was applied to upper first molar teeth of rats for 14 days. MSCs and transfected MSC injections were performed at 1st, 6th, and 11th days to the MSC and GT group rats. At the end of experiment, upper first molar teeth were prepared for genetical, scanning electron microscopy (SEM), fluorescent microscopy, and haematoxylin eosin-tartrate resistant acid phosphatase staining histological analyses. Number of total cells, number of osteoclastic cells, number of resorption lacunae, resorption area ratio, SEM resorption ratio, OPG, RANKL, Cox-2 gene expression levels at the periodontal ligament (PDL) were calculated. Paired t-test, Kruskal-Wallis, and chi-square tests were performed. Results: Transferred MSCs showed marked fluorescence in PDL. The results revealed that number of osteoclastic cells, resorption lacunae, resorption area ratio, RANKL, and Cox-2 were reduced after single MSC injections significantly (P < 0.05). GT group showed the lowest number of osteoclastic cells (P < 0.01), number of resorption lacunae, resorption area ratio, and highest OPG expression (P < 0.001). Conclusions: Taken together all these results, MSCs and GT showed marked inhibition and/or repair effects on OIIRR during orthodontic treatment on rats.

A Prospective Experimental Study on the Protective Effect of Resveratrol against Amikacin-Induced Ototoxicity in Rats.

OBJECTIVE: The purpose of this study was to evaluate the protective effect of resveratrol against amikacin-induced ototoxicity in rats by otoacoustic emission and histopathology of the cochlea. MATERIALS AND METHODS: This study was conducted with 31 Sprague Dawley adult female rats that were 20-21 weeks old and 190-245 g in weight. Before the drug administration, distortion product otoacoustic emission (DPOAE) tests were performed in both ears of each rat. The rats were divided into four groups. Group 1 (n=7) received ethanol 1cc 4%, Group 2 (n=8) received 600 mg/kg amikacin, Group 3 (n=8) received 10 mg/kg resveratrol and 600 mg/kg amikacin, and Group 4 (n=8) received 1cc resveratrol at 10 mg/kg. The drugs were administered once a day for 21 consecutive days. Control DPOAE tests were performed at the 7th, 14th, and 21st days after the administration of drugs. At the end of the study, the rats were sacrificed and their cochleae were dissected. The cochleae were evaluated for histopathologic changes. RESULTS: There was no statistically significant difference in the DPOAE measurements before the procedure between groups. The DPOAE measurements significantly decreased after the procedure in the amikacin group. There was no statistically significant difference in DPOAE measurements after the procedure in the amikacin + resveratrol, resveratrol, and ethanol groups. The histopathologic findings supported these results. CONCLUSION: We found that if resveratrol is administered with amikacin, the severity of amikacin-induced hearing loss is decreased. These findings suggest that resveratrol, a strong antioxidant, has a protective effect in amikacin ototoxicity.

Ameliorative effect of propolis on the cadmium-induced reproductive toxicity in male albino rats.

Propolis is a potent antioxidant and a free radical scavenger. The present study aimed to investigate protective effects of propolis extract on cadmium-induced testicular damage, apoptosis, HIF-1α expression and toxicity in rat's testis tissue. A total of 32 male rats were equally divided into four study groups namely, control, Cd (1mg/kg/day), Cd+propolis (50mg/kg/day) and propolis. The rats were decapitated under ketamine anesthesia and their testes tissues were removed. Serum testosterone, tissue malondialdehyde and HIF-1α levels, HIF-1α expression, apoptosis and histopathological damage scores were then compared. In the Cd group, the diameters of seminiferous tubules, tubular biopsy score of Johnsen and serum testosterone levels were decreased compared control group, but tissue HIF-1α and tissue MDA levels was higher than control group. The immunoreactivity of HIF-1α and the number of apoptotic cells were increased in Cd group. Furthermore, the propolis treated group showed an improved histological appearance in the Cd group. Thus, the results suggest that propolis acts as a potent protective agent against Cd-induced testicular toxicity in rats.

[Investigation of potentially pathogenic free-living amoebae and their in vivo pathogenicity in water supplies of Turkey]. / Türkiye'deki sularda serbest yasayan potansiyel patojen amipler ve patojenitelerinin in vivo olarak arastirilmasi.

Free-living amoebae (FLA) are found widely in soil and water in the nature. Among them in which potentially pathogenic for humans and animals are known as "potential pathogenic free-living amoebae (PPFLA)". PPFLA are characterized as the causes of clinical manifestations leading to death especially in immunosuppressed people. Four genus of PPFLA (Acanthamoeba, Naegleria, Balamuthia and Sappinia) are known to be pathogenic to humans. The aims of this study were to investigate the presence of PPFLA in the water supplies in Turkey and to determine their in vivo pathogenicity. A total of 664 water samples were collected from the ponds, rivers, streams and wells found in provinces located at different regions (central, western, eastern and southeastern regions) of Turkey. These samples were initially inoculated in the monoxenic culture media and evaluated by both microscopy and polymerase chain reaction (PCR) in terms of the presence of FLA. The samples identified as positive were then cultured in axenic media, the growth of amoebae that were confirmed microscopically, were than studied with PCR for molecular characterization. The isolates that were found positive by PCR from axenic cultures were inoculated intranasally to immunocompetent and immunodeficient (athymic) [BALB/c Rag2(-/-) gamma(c)(-/-)] BALB/c mice followed by the evaluation on the 21st day by histopathological and molecular methods to investigate their in vivo pathogenicity. In our study, 143 water samples were detected as positive in monoxenic cultures and 41 of them were detected as positive in axenic cultures. Twenty of 41 samples detected as positive in axenic culture could be continued in culture for three months. As a result of PCR using primers common to SYA, only nine have been identified from 20 samples as positive. According to the result of the PCR with specific primers, all (n= 9) were positive for Acanthamoeba sp., eight for Sappini sp. and five for Balamuthia mandrillaris, while none was observed Naegleria fowleri. Histopathologic examination revealed that both groups of mice that were infected with the nine isolates had normal brain tissue sections; but haemorrhages and mononuclear cell proliferation were determined in four immunocompetent and seven athymic animal lung sections. When the presence of parasites in tissue samples were evaluated by real-time PCR, Balamuthia was detected in at least one blood, lung, brain or nasal mucosa sample of the four immunocompetent mice, Sappinia sp. in four and Acanthamoeba sp. in seven immunocompetent mice infected with nine isolates. Additionally, seven Balamuthia sp., seven Sappinia sp. and eight Acanthamoeba sp. were detected in immunodeficient mice. In this study, B. mandrillaris and Sappinia sp. were the first isolated potentially pathogenic amoebae from water supplies located at different parts of Turkey. As a result awareness and precautions against suspicious water supplies used for drinking, daily use and swimming purposes should be treated more carefully.

Protective effects of propolis on methotrexate-induced testis injury in rat.

Propolis is an adhesive substance which is collected and used by honeybees. Propolis is a potent antioxidant and a free radical scavenger. This study was designed to determine whether propolis could protect against dysfunction and oxidative stress induced by methotrexate-induced injury in rat testis. A total of 40 male Wistar albino rats were divided into four groups: group 1 was the untreated control. On the eighth day of the experiment, groups 2 and 3 received single intraperitoneal injections of methotrexate (MTX) at 20mg/kg. Groups 3 and 4 received 100mg/kg/day propolis (by oral gavage) for 15 days by the first day of the experimental protocol. Then the rats were decapitated under anesthesia, and their testes were removed. The histopathological and biochemical analysis along with apoptosis assessment of testis tissues were compared. Immunohistochemical analysis of Heat shock protein-70 (HSP-70) and Proliferating Cell Nuclear Antigen (PCNA) were performed. The phenolic characterization of propolis was performed by Liquid chromatography-mass spectrometry (LC-MS/MS). Methotrexate caused tended to increase in malondialdehyde level and in the number of apoptotic cells; it also caused a decrease in MSTD and JTBS, PCNA and HSP-70 expression and xanthine oxidase levels in group 2. Propolis prevented the rise in malondialdehyde, xanthine oxidase levels and HSP-70 expression and improved testicular morphology and JTBS. It was found that, methorexate gives rise to serious damage in the testes and propolis is a potent antioxidant agent in preventing testicular injury.

Protective effect of propolis on methotrexate-induced kidney injury in the rat.

Objectives Propolis is a potent antioxidant and a free radical scavenger. Pharmacological induction of heat shock proteins (HSPs) has been investigated for restoring normal cellular function following an injury. In this study, effect of propolis on HSP-70 expression in methotrexate-induced nephrotoxicity and direct preventive effect of propolis in this toxicity were investigated. Material and methods A total of 40 male Wistar albino rats were divided into four groups: Group 1 was the untreated control. On the eighth day of the experiment, groups 2 and 3 received single intraperitoneal injections of methotrexate (MTX) at 20 mg/kg. Groups 3 and 4 received 100 mg/kg/day propolis (by oral gavage) for 15 d by the first day of the experimental protocol. Then the rats were decapitated under ketamine esthesia and their kidney tissues were removed. HSP-70 expression, apoptosis, and histopathological damage scores were then compared. Results MTX caused epithelial desquamation into the lumen of the tubules, dilatation, and congestion of the peritubular vessels and renal corpuscles with obscure Bowman's space. The number of apoptotic cells (p = 0.000) and HSP-70 (p = 0.002) expression were increased in group 2. Propolis prevented the rise in number of apoptotic cells (p = 0.017), HSP-70 (p = 0.000) expression, and improved kidney morphology. Conclusions It was found that methotrexate gives rise to serious damage in the kidney and propolis is a potent antioxidant agent in preventing kidney injury.

Evaluation of the effect of intranasal infiltrated botulinum toxin-A on a model of allergic rhinitis in rabbits: An Experimental Study.

INTRODUCTION: Allergic Rhinitis (AR) effects 20-40% of the global population and its prevalance increases. Medical treatment and immunotherapy could be used in AR management. But they are not definitive solution. Medical treatment must be used a long time and has side effects. Immunotherapy cannot be applied to every patient and it also takes a long time. The aim of this study is to evaluate symptomatic and histopathological effects of intranasal infiltrated Botulinum Toxin-A (Btx-A) on an animal model of AR. MATERIAL-METHOD: 15 rabbits were divided into 3 groups as control, disease and treatment. AR was formed in disease and treatment groups by intraperitoneal and intranasal ovalbumine. Allergic symptoms were observed and serum IgE levels were estimated to prove forming of AR. Btx-A was infiltrated in inferior turbinates of rabbits in treatment group. Rabbits were sacrified on 32nd day. Paranasal structures were disected and investigated histopathologically. RESULTS: Serous nasal discharge and sneezing were observed after ovalbumine applying in disease and treatment groups. Serum IgE levels on 21st day were higher than 1st day and this difference was significant statistically in disease and treatment groups. Serous discharge and sneezing decreased after Btx-A infiltration in treatment group. In histopathological examination, there were significant difference between disease and treatment group in terms of some histopathological findings. CONCLUSION: Considering the effect of Btx-A on AR in animal, it can be said that Btx-A can decrease symptoms and reorganize histopathological findings of AR.

Ameliorative effects of pentoxifylline on NOS induced by diabetes in rat kidney.

OBJECTIVES: Diabetic nephropathy (DN) is the leading cause of end-stage renal disease worldwide. The NO system has been implicated in the pathogenesis of DN. In this study, we aimed to evaluate the healing effect of pentoxifylline on NOS in STZ-induced diabetic rat's kidney. MATERIAL AND METHODS: In this study, 50 Wistar albino male rats were used. The rats were divided into five groups; Group C control; Group D only diabetes; Group D + PI and D + PII diabetes + pentoxifylline; Group P only pentoxifylline. Group DPI rats received just pentoxifylline from the beginning of the experiments. However, Group DPII rats received saline in the first month and 50 mg/kg/day of pentoxifylline for the following month. At the end of two months, NOS expressions in kidney tissue were assessed using qRT-PCR and immunohistochemistry analysis. RESULTS: At the end of the experiments, desquamation of the epithelial cells of the tubules, clear glycogen-filled distal tubules and increased number of apoptotic cells were seen in Group D. Diabetic rats' nNOS immunoreactivity had increased and eNOS and iNOS immunoreactivity had decreased; nNOS, iNOS and eNOS mRNA levels tended to decrease compared to the control group. PTX ameliorated eNOS, iNOS and nNOS protein levels and apoptotic cells, but did not affect mRNA levels. CONCLUSION: In conclusion, PTX has a healing effect on this damage by affecting NOS expression.

Effectiveness of lycopene on experimental testicular torsion.

AIM: We aimed to demonstrate the long term effectiveness of lycopene, a precursor of vitamin A, on the testes for ischemia-reperfusion injury. MATERIALS AND METHODS: Seventy male Wistar albino rats were used for this experiment. The rats were divided into seven groups. Group 1 served as the control group; group 2 was sham-operated; group 3 received 20mg/kg/day lycopene (intraperitoneally); in group 4, the right testes of rats were kept torted for 2hours and then were detorted and the animals lived for three days; in group 5, the right testes of rats were kept torted for 2hours and then were detorted and the animals lived for ten days; in group 6, the right testes of the rats were kept torted for 2hours and then detorted and the animals received 20mg/kg/day lycopene (intraperitoneally) for three days; in group 7, the right testes of the rats were kept torted for 2hours and then were detorted and the animals received 20mg/kg/day lycopene (intraperitoneally) for ten days. Lycopene was used intraperitoneally. Some of the testes tissues were used for biochemical analyses and the other tissues were used for histological procedures. The Johnsen's score was used for seminiferous tubule deterioration. The TUNEL method was utilized to show apoptosis of testicular tissue. Testosterone levels were measured from blood samples and SOD, MDA, TNF-α, IL-1ß and IL-6 measurements were recorded from tissue samples. The results were analyzed statistically. RESULTS: In groups 1, 2 and 3 there was normal testicular structure. Rats in groups 4 and 5 had damaged testicular tissues. In groups 6 and 7, in which we used lycopene, the testes were not better than those in groups 4 and 5. The MSTD and JTBS values were better in group 6, but not in group 7 among the torsion groups. As a result, MDA, SOD, TNF-α and IL-1ß were increased and serum testosterone and IL-6 levels were decreased in groups 4 and 5 compared to group 1. There was no improvement in the groups treated with lycopene for therapeutic purposes. CONCLUSION: It was shown that lycopene, as an antioxidant agent, is not effective for testicular torsion in the long term. This study can be considered as a preliminary study showing the need for further researches using different antioxidant agents to determine their long term effects in ischemia-reperfusion injuries in an appropriate experimental design.

Protective effects of grape seed extract on cadmium-induced testicular damage, apoptosis, and endothelial nitric oxide synthases expression in rats.

This study aims to evaluate the protective effect of grape seed proanthocyanidin extract (GSPE) on cadmium (Cd)-induced testicular apoptosis, endothelial nitric oxide synthases (eNOS) expression, and toxicity in rats. A total of 24 male Wistar rats were divided into four groups, namely, control, Cd (2.5 mg/kg), Cd + GSPE (100 mg/kg/day), and GSPE. Spermatogenesis and mean seminiferous tubule diameter were significantly decreased in the Cd groups. Furthermore, the GSPE-treated animals showed an improved histological appearance in the Cd group. The immunoreactivity of eNOS and the number of apoptotic cells were increased in Cd group. Our data indicate a significant reduction of terminal deoxynucleotide transferase-mediated 2'-deoxyuridine 5'-triphosphate nick end-labeling staining and a decrease in the expression of eNOS in the testes tissue of the Cd group treated with GSPE therapy. Therefore, our results suggest that GSPE acts as a potent protective agent against Cd-induced testicular toxicity in rats.

The effect of different concentrations of topical ozone administration on bone formation in orthopedically expanded suture in rats.

BACKGROUND/OBJECTIVE: The aim of this study was to investigate the effects of different concentrations of ozone (O3) therapy on bone regeneration in response to an expansion of the inter-premaxillary suture in rats. MATERIALS AND METHODS: Forty-eight Wistar rats were randomly divided into four groups (n = 12). In groups I, II, and III, 1ml of O3 at 10, 25, and 40 µg/ml was injected at the premaxillary suture, respectively. In group IV (control group), 1ml of saline solution was injected at the same point during the expansion procedure for 5 days. Bone regeneration in the suture was evaluated histomorphometrically. The area of new bone and fibrotic area, the number of osteoblasts and osteoclasts, and the amount of vascularity were measured and compared. The density of the newly formed bone in the expansion area was measured by using cone beam computed tomography. Data were analyzed using the Kruskal-Wallis one-way analysis of variance and post hoc Student-Newman-Keuls tests. RESULTS: New bone area, fibrotic area, osteoblast and osteoclast numbers, and the amount of vascularity were significantly higher in experimental groups compared with the control group (P < 0.001). The density of newly formed bone (P < 0.001), new bone formation (P = 0.009), number of capillaries (P < 0.001), number of osteoclasts (P = 0.016), and number of osteoblasts (P < 0.001) in the maxillary sutures were highest in the 25 µg/ml O3 group compared with the other experimental groups and control group. CONCLUSIONS/IMPLICATIONS: The application of O3 therapy can stimulate bone regeneration in an orthopedically expanded inter-premaxillary suture during both the expansion and retention periods.

Therapeutic effects of pentoxifylline on diabetic heart tissue via NOS.

OBJECTIVE: Diabetes mellitus causes a decrease in cardiac output, arterial blood pressure, and heart rate. In this study, we aimed to investigate, at the molecular level, the effect of nitric oxide synthase (NOS) on heart pathology in type 1 diabetes and look at the therapeutic effect of pentoxifylline on this pathology. METHODS: In this experimental study, 50 Wistar albino male rats were used. The rats were divided into 5 groups: group C, control; group D, only diabetes; group D+PI and D+PII, diabetes + pentoxifylline; group P, only pentoxifylline. Group D+PI rats received 50 mg/kg/day pentoxifylline over two months. However, group D+PII rats received saline in the first month and 50 mg/kg/day of pentoxifylline over the following month. At the end of two months, NOS expressions in heart tissue were assessed through immunohistochemistry analysis. The data were compared by one-way ANOVA. RESULTS: At the end of the experiments, there was increased cytoplasmic vacuolization, myofibrillar loss, cytoplasmic eosinophilia, and degeneration of cardiomyocytes; nNOS and iNOS expressions in group D decreased compared with that in group C. In group D+PI and group D+PII, nNOS and iNOS expressions improved compared with group D. CONCLUSION: As a result, we found that diabetes, a known chronic disease, causes serious damage in heart tissue. NOS plays a role in this damage, and pentoxifylline aided in improving nNOS and iNOS expression in this damage.

Nitric oxide synthase in diabetic rat testicular tissue and the effects of pentoxifylline therapy.

Diabetes is known to be associated with erectile dysfunction, retrograde ejaculation, level of testicular hormone, and a decrease in semen quality, respectively. In this project, we aimed to investigate at the molecular level, the effects of NOS on testes pathology in diabetes and examine the effects of pentoxifylline on healing. A total of 50 Wistar albino male rats were divided into five groups: Group I control; Group II only diabetes; Group III and IV diabetes + pentoxifylline; Group V only pentoxifylline. Group III rats received 50 mg/kg/day pentoxifylline during two months. In comparison, Group IV rats received saline in the first month followed by 50 mg/kg/day of pentoxifylline for the following month. NOS expression in testicular tissue was assessed using qRT-PCR, western blot, and immunohistochemistry. The mean seminiferous tubule diameter, Johnsen's testicular biopsy score, and serum testosterone levels decreased compared to controls. In contrast, the number of apoptotic cells, the levels of nNOS, iNOS and eNOS mRNA, and protein increased when compared to the control. Upon pentoxifylline therapy NOS decreased suggesting that it contributes to this damage and treatment with pentoxifylline may be effective in reversing this damage.

Cardioprotective effect of metformin against doxorubicin cardiotoxicity in rats.

OBJECTIVE: The clinical use of doxorubicin, which is a strong antineoplastic agent, is limited due to its cardiotoxic side effects. Metformin is a drug with antihyperglycemic effects, and it has been shown to have a cardioprotective effect on left ventricular function in experimental animal models of myocardial ischemia. The present study investigated the cardioprotective effect of metformin in rats with doxorubicin cardiotoxicity. METHODS: Wistar albino rats were used in the study. Forty male, 10-week-old Wistar albino rats were randomly divided four groups. The control group rats were intraperitoneally administered saline solution twice a week, four doses in total. The doxorubicin group rats received doxorubicin (4 mg/kg, twice a week, cumulative dose: 16 mg/kg) intraperitoneally. The metformin group rats received metformin (250 mg/kg/day, every day for 14 days) via gavage. The doxorubicin + metformin group rats received doxorubicin and metformin at the same dose. Left ventricular functions were evaluated by using M-mode echocardiography one day after the last dose of doxorubicin. Heart tissue samples were histopathologically examined. Cardiomyocyte apoptosis was detected using in situ terminal deoxynucleotide transferase assay (TUNEL). Serum brain natriuretic peptide and C-type natriuretic peptide levels were measured. Catalase, superoxide dismutase, glutathione peroxidase, and tumor necrosis factor alpha levels were analyzed in the heart tissue. The assumptions of equality of variances and normal distribution were checked for all variables (Shapiro-Wilk test and Q-Q graphics).To identify intergroup differences, one-way variant analysis or the Kruskal-Wallis test was used. A p<0.05 value was accepted as statistically significant. RESULTS: Our results showed that doxorubicin treatment caused significant deterioration in left ventricular functions by echocardiography, histological heart tissue damage, and increase in cardiomyocyte apoptosis. Doxorubicin + metformin group showed protection in left ventricular function, elimination of histopathologic change, and reduced of cardiomyocyte apoptosis. CONCLUSION: The present study provided evidence that metformin has cardioprotective effects against doxorubicin cardiotoxicity.