Histopathological and biomechanical evaluation of tenocyte seeded allografts on rat Achilles tendon regeneration.

Tendon injuries in humans as well as in animals' veterinary medicine are problematic because tendon has poor regenerative capacity and complete regeneration of the ruptured tendon is never achieved. In the last decade there has been an increasing need of treatment methods with different approaches. The aim of the current study was to improve the regeneration process of rat Achilles tendon with tenocyte seeded decellularized tendon matrices. For this purpose, Achilles tendons were harvested, decellularized and seeded as a mixture of three consecutive passages of tenocytes at a density of 1 × 10(6) cells/ml. Specifically, cells with different passage numbers were compared with respect to growth characteristics, cellular senescence and collagen/tenocyte marker production before seeding process. The viability of reseeded tendon constructs was followed postoperatively up to 6 months in rat Achilles tendon by histopathological and biomechanical analysis. Our results suggests that tenocyte seeded decellularized tendon matrix can significantly improve the histological and biomechanical properties of tendon repair tissue without causing adverse immune reactions. To the best of our knowledge, this is the first long-term study in the literature which was accomplished to prove the use of decellularized matrix in a clinically relevant model of rat Achilles tendon and the method suggested herein might have important implications for translation into the clinic.

Carvacrol partially reverses symptoms of diabetes in STZ-induced diabetic rats.

Little is known about the protective effects of carvacrol on the symptoms of streptozotocin induced diabetes in rats. Hence, this present study was designed to evaluate the protective effect of the strong antioxidant, carvacrol, on the symptoms of streptozotocin induced diabetes in rats. Carvacrol at the doses of 25 and 50 mg/kg body weight were orally administered to diabetic rats for a period of 7 days after the onset of diabetes. Food-water intake and body weight changes were daily recorded. Biochemical parameters such as serum glucose, insulin, total cholesterol, alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase were measured. Although treatment of diabetic rats with oral administration of carvacrol resulted in a slight reduction in serum glucose level and significant reduction in serum total cholesterol, alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase in comparison with diabetic control rats, there were no significant differences in serum insulin levels, food-water intake values and body weight changes. Despite the inadequacy of carvacrol on diabetes treatments, it was determined to have at least a partially protective role on liver enzymes.

Inhibiting inducible nitric oxide synthase with rutin reduces renal ischemia/reperfusion injury.

BACKGROUND: Nitric oxide (NO) seems to play an important role during renal ischemia/reperfusion (I/R) injury. We investigated whether rutin inhibits inducible nitric oxide synthase (iNOS) and reduces 3-nitrotyrosine (3-NT) formation in the kidneys of rats during I/R. METHODS: Wistar albino rats were nephrectomized unilaterally and, 2 weeks later, subjected to 45 minutes of left renal pedicle occlusion followed by 3 hours of reperfusion. We intraperitoneally administered L-N6-(1-iminoethyl)lysine (L-NIL; 3 mg/kg) for 30 minutes or rutin (1 g/kg) for 60 minutes before I/R. After reperfusion, kidney samples were taken for immunohistochemical analysis of iNOS and 3-NT. We measured plasma nitrite/nitrate and cyclic guanosine monophosphate (cGMP) to evaluate NO levels. RESULTS: Ischemia/reperfusion caused plasma cGMP to increase significantly. Similarly, plasma nitrite/nitrate was elevated in the I/R group compared with the control group. Histochemical staining was positive for iNOS and 3-NT in the I/R group. Pretreatment with L-NIL or rutin significantly mitigated the elevation of plasma cGMP and nitrite/nitrate. These changes in biochemical parameters were also associated with changes in immunohistochemical appearance. Pretreatment with L-NIL or rutin significantly decreased the incidence and severity of iNOS and 3-NT formation in the kidney tissues. CONCLUSION: Our findings suggest that high activity of iNOS causes renal I/R injury, and that rutin exerts protective effects, probably by inhibiting iNOS.

Persistent organochlorine pesticide residues in water and sediment samples from Lake Manyas, Turkey.

The extent of organochlorine pesticides (OCPs) contamination in Manyas Lake (Bird Lake) and its tributary rivers and streams which is situated near the South-eastern coasts of the Marmara Sea in Turkey was investigated. Residues of OCPs in surface water and sediment samples were analysed by gas chromatography equipped with electron capture detector. The total OCPs concentration (sigmaOCPs) in water and sediment samples ranged from 1.43 to 8.6 microg l(-1) and from 17.05 to 39.14 ng g(-1), respectively. According to the concentrations and detection frequencies, HCHs, DDTs, heptachlor and heptachlor epoxide were the most dominant compounds among the OCPs. The results showed that Manyas Lake was still contaminated by organochlorine pesticides and their residues despite bans on the production and usage over a long time.

Gene expression of tendon collagens and tenocyte markers in long-term monolayer and high-density cultures of rat tenocytes.

As a result of repeated movement, tendons are functionally open to traumas. According to this situation, tenocytes have already been used for tissue engineering therapies. It has been reported that long-term monolayer (ML) culture of tenocytes may lead to a phenotypic drift within passages. Depending on our previously published work, it is clearly demonstrated that high-density (HD) culture improves cell growth and differentiation of tenocytes. However, it is not yet established if HD favors the differentiated state during long-term culture. Therefore, we compared the differences in gene expression of tendon collagens and tendon markers of tenocytes from long-term ML and HD culture conditions by quantitative, real-time polymerase chain reaction (QRT-PCR) for over a period of 3 weeks. COLI, COLIII, COLV, Scx, and Tnmd were target genes as the major matrix constituents of tendons as well as being involved in matrix integrity and tenocyte phenotype. According to our results, tenocytes in HD culture synthesized less amounts of COLIII, COLV, and Tnmd, and dependent on the investigation time point, higher amounts of Scx. We consider that tenocytes produced in HD culture system may not provide sufficient efficiency during tissue engineering approaches. By the fact that most molecules showed significantly higher expression profiles in ML culture condition, it is suggested that culture and passage in ML should be taken into consideration for further tissue engineering approaches to maintain a phenotype with less amount of drift.

Evaluation of spatial and temporal changes in biomarker responses in the common carp (Cyprinus carpio L.) for biomonitoring the Meriç Delta, Turkey.

The aim of this study is to examine the activity of several biomarkers in carp, Cyprinus carpio L., to determine their response to xenobiotics, such as organochlorine pesticides (OCPs), in the Meriç Delta. Fish were collected from contaminated sites and from areas regarded as relatively less contaminated in four sampling periods. Hepatic glutathione S-transferase (GST), carboxylesterase (CaE), lactate dehydrogenase, aspartate aminotransferase, and acid phosphatase activities were measured as biomarkers in the fish. For all fish, the condition factor and hepatosomatic index, were calculated to determine the condition of the fish. The results of this study indicated that the mean GST activity showed an increase in fish from the Meriç-Ergene junction site and a decrease in Enez site with respect to fish from Meriç site. Furthermore, the study shows that spatial and temporal changes of biomarkers such as GST and CaE might be useful for the assessment of environmental contamination in the Meriç Delta.

Vitamin C coadministration augments bisphenol A, nonylphenol, and octylphenol induced oxidative damage on kidney of rats.

The aim of this study was to investigate whether bisphenol A (BPA), nonylphenol (NP), and octylphenol (OP) induce oxidative stress on the kidney tissue of male rats and whether coadministration of vitamin C, an antioxidant, can prevent any possible oxidative stress. The Wistar male rats were divided into seven groups, including control, BPA, NP, OP, BPA+C, NP + C, OP +C. BPA, NP, and OP (25 mg/kg/day) was administered alone; vitamin C (60 mg/kg/day) was administered along with BPA, OP, and NP to the rats for 50 days. There was a decrease in serum concentration of blood urea nitrogen (BUN) in NP and OP groups compared with control group. Vitamin C coadministration with BPA, NP, and OP did not produce significant increase in BUN concentration in BPA +C, NP+ C, and OP + C group as compared with BPA, NP, and OP groups, respectively. The lowest serum creatinine activity and the highest lactate dehydrogenase (LDH) activity was present in kidney of BPA+C, NP+C and OP+C groups compared with BPA, NP, and OP groups. The malondialdehyde (MDA) levels were significantly higher while glutathione (GSH) levels were lower in treatment groups than controls. Furthermore, an increase was observed in MDA levels whereas a decrease was observed in GSH levels in BPA+ C, NP + C, and OP+ C groups compared with BPA, NP, and OP groups, respectively. These finding are in accordance with immunohistochemical staining of MDA and GSH. Histopathological examination of the kidneys of rats in BPA, OP, NP, BPA+ C, NP + C, and OP+ C groups revealed necrotic lesions, congestion, and mononuclear cell infiltration. In conclusion BPA, NP, and OP might induce oxidative damage in kidney of rats. In addition, coadministration of vitamin C with BPA, NP, and OP to male rats augments this damage in the kidney of male rats.

Influence of vitamin C on bisphenol A, nonylphenol and octylphenol induced oxidative damages in liver of male rats.

The purpose of this study was to investigate whether bisphenol A (BPA), nonylphenol (NP) and octylphenol (OP) induce oxidative stress in the liver of male rats and co-administration of vitamin C can prevent any possible oxidative stress. Wistar male rats were divided into seven groups (vehicle, BPA, NP, OP, BPA+C, NP+C, OP+C). BPA, OP and NP groups (25 mg kg(-1)day(-1)) were administered orally to rats three times a week for 50 days. In BPA+C, NP+C, OP+C groups, vitamin C (60 mg kg(-1)day(-1)) was administered along with BPA, OP and NP (25 mg kg(-1)day(-1)) treatments. Aspartate transaminase (AST), alanine transaminase (ALT), lactate dehydrogenase (LDH), thiobarbituric acid-reactive substances (TBARS) were increased, glutathione (GSH) levels were decreased in treatment groups. AST, ALT, LDH and TBARS levels were increased whereas GSH levels were decreased in BPA+C, NP+C and OP+C groups compared to BPA, NP, and OP groups, respectively. Hepatic necrosis and congestion were observed in livers of rats treated. In conclusion, the present results demonstrate that BPA, NP, and OP cause oxidative damage by disturbing the balance between ROS and antioxidant defenses system in liver of male rats. Vitamin C co-administration along with BPA, NP, OP aggravates the damage in liver of male rats.

The effects of maternal exposure to food additive E341 (tricalcium phosphate) on foetal development of rats.

E341 (tricalcium phosphate) (TCP) is commonly used as a food additive and also as a nutritional supplement. To evaluate the possible developmental effects, female Wistar rats were treated with E341 (TCP) by oral gavage during pregnancy. There were three groups of each containing five rats. Rats in Groups I-III were fed with standard diet, oil and E341 (TCP) 175mg/kg body weight during gestation days (GD 0-20) respectively. We assessed foetal body lengths and weights and also made morphometric examination of placenta and umbilical cord. The placental weights of E341 (TCP) group (Group III) were found to be decreased statistically. According to skeletal stainings of foetuses, lengths of left ulna (28.3%), right femur (29.8%), left femur (34.9%) and diameter of the skull of y-axis were significantly decreased (12.3%) in E341 (TCP) treatment groups. There was an increase in trans-umbilical diameter in treatment group (14%). This is the first study in which developmental effects of E341 (TCP) have ever evaluated. The results suggest that prenatal development of rats during gestation is sensitive to E341 (TCP) exposure.

Protective effect of rutin on the ischemia/reperfusion induced damage in rat kidney.

Reactive oxygen species (ROS) are suggested to participate in ischemia/reperfusion (I/R) injury in the kidney. This study was designed to investigate the effect of rutin, a bioflavonoid, in I/R induced renal injury. Wistar albino rats were unilaterally nephrectomized, and 2 wk later they were subjected to 45 min of left renal pedicle occlusion followed by 3h of reperfusion. Either rutin (1g/kg) or saline was administrated (i.p.) 1h prior to ischemia. At the end of the reperfusion period, kidney samples were taken for determination of renal malondialdehyde (MDA) and glutathione (GSH) levels, manganese-superoxide dismutase (MnSOD) activity and histological examination. Serum creatinine, blood urea nitrogen (BUN), and lactate dehydrogenase (LDH) concentrations were measured for the evaluation of renal function. I/R caused a significant decrease in GSH level and MnSOD activity, which was accompanied by a significant increase in MDA level of kidney tissues. Similarly, serum BUN and creatinine levels, as well as LDH were elevated in the I/R group compared with the control group. Pretreatment of rats with rutin (1g/kg/ i.p.) significantly attenuated renal dysfunction, reduced elevated MDA levels, and restored the depleted MnSOD activity and GSH levels. These beneficial changes in the biochemical parameters were also associated with parallel changes in histopathological appearance. These findings suggest that ROS play a causal role in I/R induced renal injury, and that rutin exerts renal-protective effects, probably by inhibiting ROS and antioxidant activities.

Pro-oxidant effect of vitamin C coadministration with bisphenol A, nonylphenol, and octylphenol on the reproductive tract of male rats.

This study was performed to investigate whether bisphenol A (BPA), nonylphenol (NP), and octylphenol (OP) induce oxidative stress on the reproductive tract of male rats and if coadministration of vitamin C can prevent any possible oxidative stress. Wistar male rats were divided into seven groups as control (vehicle; olive oil), BPA, NP, OP, BPA+C, NP+C, and OP+C. BPA, OP, and NP groups (25 mg/kg/day) were administered orally to rats three times a week for 45 days. In BPA+C, NP+C, and OP+C groups, vitamin C (60 mg/kg/day) was administered orally along with BPA, OP, and NP (25 mg/kg/day) treatments. Malondialdehyde (MDA) appeared at significantly higher concentrations in BPA-, NP-, and OP-treated groups, when compared to control group. No significant decrease was observed in testes MDA levels of vitamin C coadministrated groups, compared with BPA, NP, and OP treatment groups. Decreased levels of reduced glutathione (GSH) were found in testes of BPA-, NP-, OP-treated rats. No significant increase was observed in testes GSH levels of BPA+C, NP+C, and OP+C groups, compared with BPA, NP, and OP treatment groups. Histological examination showed that vitamin C coadministrated groups had much more congestion areas, atrophy, and germinal cell debris in testes than those observed in other groups. Abnormal sperm percentages of BPA, BPA+C, NP+C, and OP+C groups were increased. In conclusion, the present results demonstrated that BPA, NP, and OP generate reactive oxygen species that cause oxidative damage in testes of rats. Coadministration of vitamin C aggravates this damage.

The protective effects of ascorbic acid against renal ischemia-reperfusion injury in male rats.

There is increasing evidence to suggest that toxic oxygen radicals play an essential role in the pathogenesis of ischemia/reperfusion (I/R) injury in the kidney. This study was designed to investigate the effects of ascorbic acid (AA) in I/R-induced renal injury in rats. Thirty two male Sprague-Dawley rats were divided equally into four groups: group 1 (control; dissection of the right renal pedicle without nephrectomy), group 2 (sham operated; unilateral nephrectomy), group 3 (I/R; unilateral nephrectomy + I/R); and group 4 (AA+I/R; unilateral nephrectomy and I/R treated with ascorbic acid, 250mg kg(-1) i.p., for one hour prior to ischemia). On the 15th day following nephrectomy, groups 3 and 4 were subjected to 45 min of renal pedicle occlusion followed by 3 h of reperfusion. At the end of the treatment period, kidney samples were taken for histological examination or determination of the renal malondialdehyde (MDA) and glutathione (GSH) levels. Serum creatinine, blood urea nitrogen (BUN), and lactate dehydrogenase (LDH) concentrations were measured for the evaluation of renal function. I/R caused a significant decrease in GSH level, which was accompanied with a significant increase in MDA level of kidney tissues. Similarly, serum BUN and creatinine levels, as well as LDH, were elevated in the I/R group as compared to the control group. In group four, AA treatment reversed all the changes in these biochemical indices, as well as histopathological alterations normally induced by I/R. The findings imply that reactive oxygen species play a causal role in I/R-induced renal injury, and that AA exerts renoprotective effects, probably by radical scavenging and antioxidant activities.

Anzer honey prevents N-ethylmaleimide-induced liver damage in rats.

N-ethylmaleimide (NEM) is a sulphydryl blocker which impairs the sulphydryl dependent antioxidant system (mainly glutathione) in the body by alkylating endogenous sulphydryls. This study was designed to investigate the effects of Anzer honey on NEM-induced liver injury in rats. Thirty female Wistar albino rats were divided equally into three groups. Group 1: control; Group 2: NEM; Group 3: Anzer honey+NEM. NEM (0.075mg kg(-1)) was given to both group 2 and 3 administered subcutaneously (s.c.) for 30 days. The animals in the Anzer honey+NEM group were treated with Anzer honey at a dose of 0.275g kg(-1), (p.o.) at 1h prior to every NEM injection. At the end of the 30 day treatment period, liver samples were taken for determination of the glutathione levels and histological examination. NEM treatment alone caused a significant reduction of the liver glutathione levels in group 2. Furthermore, NEM treatment caused congestion and mononuclear cell infiltration in the liver when compared to the control group. In group 3, Anzer honey treatment reversed all the changes in glutathione level, as well as histopathological alterations, normally induced by NEM. The findings imply that depletion of glutathione concentration plays a causal role in NEM-induced liver injury, and that the hepatoprotective effect of Anzer honey may be mediated through sulfhydryl-sensitive processes. They further imply that it may also possess antioxidant properties.

The effect of vitamin C on bisphenol A, nonylphenol and octylphenol induced brain damages of male rats.

Bisphenol A (BPA), nonylphenol (NP) and octylphenol (OP) are endocrine-disrupting chemicals that has been shown to exert both toxic and estrogenic effects on mammalian cells. The aim of this study was to investigate if BPA, NP and OP induce oxidative stress on the brain tissue of male rats and if co-administration of vitamin C, an antioxidant, can prevent any possible oxidative stress. The male rats were divided into seven groups as control (vehicle), BPA, NP, OP, BPA+C, NP+C, OP+C. BPA, OP and NP (25 mg/(kg day)) were administrated orally to male Wistar rats for 45 days. In vitamin C co-administration groups (BPA+C, NP+C, OP+C), vitamin C (60 mg/(kg day)) were administrated orally along with BPA, OP and NP (25 mg/(kg day)) treatments. The rats in the control group received olive oil orally. The final body and absolute organ weights of treated rats did not show any significant difference when compared with the control group. Also, there were no significant difference in relative organ weights of BPA, NP, OP, BPA+C and NP+C groups when compared with control group. Only, relative organ weights were increased significantly in OP+C group compared with control group. Decreased levels of reduced glutathione (GSH) were found in the brains of BPA, NP, OP treated rats. The end product of lipid peroxidation, malondialdehyde (MDA), appeared at significantly higher concentrations in the BPA, NP, and OP treated groups when compared to the control group. On the other hand, there were no changes in the brain MDA and GSH levels of BPA+C, NP+C and OP+C groups compared with BPA, NP and OP treatment groups, respectively. In histopathologic examination, the vitamin C co-administrated groups had much more hyperchromatic cells in the brain cortex than that observed in the groups treated with only BPA, NP, and OP. The results of this study demonstrate that BPA, NP and OP generate reactive oxygen species that caused oxidative damage in the brain of male rats. In addition, vitamin C co-administration along with BPA, NP, and OP aggravates this oxidative damage in the brain of rats.

The effect of sub-acute and sub-chronic exposure of rats to the glyphosate-based herbicide Roundup.

Roundup is a glyphosate-based herbicide that includes 78.5% glyphosate and surfactant at lower toxic concentrations. Glyphosate is an organophosphorated non-selective agrochemical widely used in many countries including Turkey and acts after the sprout in a systemic way. The objective of this study was to analyze toxic effects of the herbicide Roundup in rat liver. Animals were treated with 56mg/kg (p.o.) and 560mg/kg (p.o.) of Roundup (78% glyphosate+surfactant) each day, during 5 and 13 weeks. Hepatotoxicity was monitored by quantitative analysis of the serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) activities and measured amount of serum lipoprotein (LDL, HDL), total cholesterol and creatinine were used as the biochemical markers of liver damages. Besides the biochemical analysis, we also investigated liver tissues histopathologically. Sub-chronic treatment, starting from the low and high doses of Roundup, it was observed that there were mild effects on activity of ALT, AST and LDH enzymes indicating the hepatic toxicity induced by Roundup. It was found that the mild effects were different on the enzymes in male and female rats of treatment groups. Also it was found some difference in serum lipoprotein (LDL, HDL) and t-cholesterol. There was no difference creatinine value between control and treatment groups but it was observed that degenerative formation such as mononuclear cell infiltration and congestion of the liver tissues of treatment groups.

Characterization of type I, III and V collagens in high-density cultured tenocytes by triple-immunofluorescence technique.

The purpose of this study is to examine the intracellular distribution of collagen types I, III and V in tenocytes using triple-label immunofluorescence staining technique in high-density tenocyte culture on Filter Well Inserts (FWI). The tenocytes were incubated for 4 weeks under monolayer conditions and for 3 weeks on FWI. At the end of the third week of high-density culture, we observed tenocyte aggregation followed by macromass cluster formation. Immunofluorescence labeling with anti-collagen type I antibody revealed that the presence of collagen type I was mostly around the nucleus. Type III collagen was more diffused in the cytoplasm. Type V collagen was detected in fibrillar and vesicular forms in the cytoplasm. We conclude that, the high-density culture on FWI is an appropriate method for the production of tenocytes without loosing specialized processes such as the synthesis of different collagen molecules. We consider that the high-density culture system is suitable for in vitro applications which affect tendon biology and will improve our understanding of the biological behavior of tenocytes in view of adequate matrix structure synthesis. Such high-density cultures may serve as a model system to provide sufficient quantities of tenocytes to prepare tenocyte-polymer constructs for tissue engineering applications in tendon repair.

The effect of Trifolium, Raphanus, and Cistus pollen grains on some blood parameters and mesentery mast cells.

Three kinds of pollen taxa belonging to 3 families (Fabaceae--Trifolium spp., Brassicaceae--Raphanus spp. and Cistaceae--Cistus spp.) and commonly collected by honeybees were fed to mature male rats separately, in the form of 60 mg/animal/day for a 30-day period. The objective of this study was to investigate any positive effects or possible side effects of the use of pollen on the immune system. This was achieved through blood analysis and cell count on blood, hemoglobin, erythrocyte and immune system cells. The cell concentration of mast cells, degranulization and cell localization were investigated in prepared mesentery tissue samples. Histological investigations of the stomach and duedenum sections of pollen-fed rats were carried out to learn the reason for eosinophil gastroenteritis in the alimentary canal. The eosinophil and lymphocyte levels of rats fed with pollen of Trifolium spp., Raphanus spp., and Cistus spp. were observed to have increased blood cell counts, while neutrophil and monocyte levels decreased; different values were found in basophil leucocytes between the pollen groups. Differing reductions in mesentery mast cell concentration, degranulization and cell localization were found. Within the three separate pollens, the rats having been fed with Cistus spp. pollen were observed to have higher blood lymphocyte, eosinophil, hemoglobin and hematocrit values than those fed with the others, as well as low mesentery mast cell concentration. Hemoglobin values were determined to increase at a proportion of between 10.0-11.3%. No difference was found in other blood parameters. The fat proportion of the male rats fed with the three taxa was between 4.03-8.75%, while that for protein proportion was between 16.11-24.25%. Male rats receiving these taxa did not experience allergic reactions and it is possible to argue that the low protein and fat content of these pollens have a strengthening effect on the immune systems by the increase in lymphocyte content and the amount of hemoglobin leads to an increase of oxygen transport capacity in the tissues.

Protective effect of Anzer honey against ethanol-induced increased vascular permeability in the rat stomach.

The purpose of this study was to determine the protective effect of Anzer honey on ethanol-induced increased vascular permeability in rats. Evan's Blue (EB) dye, administered intracardiacly and extravasation of EB into the stomach, served as an indicator of vascular permeability following exposure to alcohol. Ethanol was given orally to the ethanol group for 90 days, and N-etylmaleimide (NEM) was given subcutaneously to the NEM group, and we observed increased extravasation of EB in the stomach in both groups. For this reason, we used NEM as a positive control for ethanol. Anzer honey, which contains 25.44 mg/g ascorbic acid, was given to the honey+ethanol group orally 30 min before beginning the 90-day ethanol administration. The mean amount of EB that leaked into the stomach of rats in the ethanol group and the NEM group was higher than that of the control group. Furthermore, if compared to the control, EB values in the stomachs were significantly reduced when receiving honey before administration of ethanol in rats. Histopathologically, the incidence and severity of gastric mucosal congestion were significantly reduced in the honey+ethanol group when compared to the ethanol group. These result indicate that Anzer honey is able to protect the stomach of the rat against ethanol-induced increased vascular permeability, which may be correlated with the ascorbic acid content.