Effect of Bacillus licheniformis and Bacillus subtilis Supplementation of Ewe's Feed on Sheep Milk Production and Young Lamb Mortality

The purpose of this pilot study was to evaluate under field conditions the effect of a probiotic containing Bacillus licheniformis and Bacillus subtilis on young lamb mortality and sheep milk production when administered in the late pregnancy and lactation feed of ewes. In a sheep farm, two groups of milking ewes with identical genetic material, management, nutrition, health status and similar production characteristics were formed. One group (46 ewes) served as control, while the other one (48 ewes) served as a probiotic‐treated group. Both groups of ewes received a similar feeding regiment, but the ewes of the second group were additionally offered a probiotic product containing B. licheniformis and B. subtilis (BioPlus 2B, Chr. Hansen, Denmark) at the approximate dose of 2.56 × 10⁹ viable spores per ewe per day. Lamb mortality during the 1.5 months suckling period, and milk yield during the 2 months of milk collection for commercial purposes have been recorded. In the non‐treated control group, 13.1% mortality was observed versus 7.8% in the probiotic‐treated group (P = 0.33), with mortality being mainly due to diarrhoea. Microbiological examination of diarrhoeic faeces from some of the dead lambs in both groups revealed the presence of Escherichia coli. The average daily milk yield per ewe was significantly lower in the control group (0.80 l) than that in the probiotic‐treated group (0.93 l) (P < 0.05). Fat and protein content of milk in ewes that received probiotics was significantly (P < 0.05) increased compared with untreated ewes. It was concluded that supplementing ewe's feed with probiotics may have beneficial effect on subsequent milk yields, fat and protein content.     

Rutin,an antioxidant flavonoid,induces glutathione and glutathione peroxidase activities to protect against ethanol effects in cadmium‐induced oxidative stress in the testis of adult rats

Exposure to cadmium (Cd) reduces sperm quality and induces oxidative stress in the testis. Rutin is an effective antioxidant flavonoid. We studied the effect of ethanol (EtOH, 5 g/kg b.wt.) intake on Cd (50 mg/kg b.wt.)‐induced testicular toxicity with or without RUT pre‐treatment (25, 50, 100 mg/kg b.wt.) in rats. At the end of the 15‐day oral treatment, co‐treatment with EtOH decreased the activities of glutathione (GSH), GSH‐peroxidase and superoxide dismutase resulting to slight increase in the testicular MDA level compared to Cd‐treated rats. The Cd+EtOH animals had higher levels of abnormal spermatozoa, decreased epididymal sperm number and serum testosterone levels (p < .05) compared to the Cd‐treated animals. Rutin co‐administration protected against the EtOH effects in a dose‐dependent manner, with the Cd+EtOH+50 mg/kg RUT‐ and Cd+EtOH+100 mg/kg RUT‐treated animals having higher GSH and GSH‐Px activities beyond the control values (p < .05). In a supplementary study, animals treated daily with RUT alone (25, 50, 100 mg/kg b.wt.) for 15 days dose‐dependently increased testicular GSH‐peroxidase and GSH activities by 9.38%, 31.25%, 56.25% and 7.14%, 32.14%, 60.71%, respectively, compared to control values. Therefore, RUT induces GSH and GSH‐Px activities to protect against Cd+EtOH‐induced testis oxidative stress in rats.     

Humeral skeletal development and plasma constituent changes in fetuses of ewes maintained on a low calcium diet from 60 days of gestation

Summary The objective of this study was to evaluate the effects of a long-term, low-calcium diet on fetal calcium metabolism and fetal skeleton skeleton development in ewes. Eleven pregnant sheep were assigned to two groups, fed either a diet low in calcium (0.26% total dry matter) or normal in calcium (0.8% total dry matter) for 2 months, starting at 60 days gestational age. The ewes fed the low calcium diet showed lower plasma levels of calcium and higher plasma levels of hydroxyproline, parathyroid hormone, and 1,25 (OH)₂D compared with the ewes fed the normal calcium diet. There were no differences in these variables between the two groups of fetuses. These observations suggest that the plasma components of calcium homeostasis measured in the fetal lamb in the present study are independent of the ewe and are not significantly affected by the presence of lowere maternal calcium for many weeks during pregnancy. Despite the ability of the fetus of the ewe on the low calcium diet to maintain relatively normal circulating plasma components of calcium homeostasis, long-term maternal hypocalcemia delayed fetal skeletal ossification as shown by histological examination of the fetal humerus. The fetal humerus from low calcium-fed ewes showed a lower proportion of bone versus cartilage (45.6±5.9 versus 57.4±4.6%, mean ±SD) lower ash content (15.4±1.5 versus 17.4±1.0%), and lower specific gravity (1.19±0.2 versus 1.22±0.02) (P<0.05) than the humerus from fetuses of normal calcium-fed ewes. This study shows that the long-term calcium intake of the ewe does affect fetal skeletal development, despite a lack of observable effects on fetal plasma concentrations of calcium or known calcium regulating hormones such as 1,25(OH)₂D or parathyroid hormone.     

Effects of Three Anthelmintic Regimes on Milk Yield of Ewes and Growth of Lambs

Forty ewes were allocated into one of four groups (n = 10) and were treated with albendazole (ALB) (3.8 mg kg⁻¹) before lambing (group A), with ALB before and after lambing (group B), with moxidectin (MXD) (0.2 mg kg⁻¹) before lambing (group C) or were untreated controls (group D). Counts of nematode eggs in faeces and coprocultures were carried out during the study, as well as ewes’ milk yield measurements and lamb weighings. Pre‐treatment mean eggs per gram (epg) counts were 640, 715, 625 and 630 for groups A, B, C and D, respectively (P > 0.05); respective counts 21 days after treatment were 5, 0, 0 and 690 epg, whilst 70 days after treatment they were 380, 145, 40 and 1120 epg. Mean lactation milk yield was 3527.5, 3893.5, 3786.4 and 3285.9 ml for groups A, B, C and D, respectively; no significant difference was evident among the four groups in milk yield collected during the suckling period, although subsequently, group B or C ewes yielded significantly more milk than controls (P < 0.05). Mean birthweight of lambs were 3.56, 3.45, 3.59 and 3.26 kg for groups A, B, C and D, respectively (P = 0.045); subsequently, lambs from treated ewes were significantly heavier than lambs from control animals (P < 0.001). We conclude that anti‐parasitic treatment during the last month of pregnancy contributed to an increased birthweight of lambs of treated ewes, whilst the cumulative effect of two doses of ALB or the long persistent efficacy of MXD provided a longer protection of animals against new parasitic infections and contributed to a lactation persistence.     

Protective effect of thymoquinone against testicular torsion induced oxidative injury

We aimed to determine the protective effects of thymoquinone (TQ), against ischaemia–reperfusion (I/R) injury in the testis tissue of rats. Twenty‐seven male Wistar albino rats were randomly divided into three equal groups as follows: Group I, sham group; Group II, torsion group; and Group III, torsion + thymoquinone group. The ischaemia period was 2 h, and orchiectomy was performed after 30 min of detorsion. Testis tissue sections were analysed with the terminal transferase mediated dUTP‐nick end labelling (TUNEL) assay to determine in situ apoptotic DNA fragmentation. Additionally, Caspase 3 and Bax proteins were analysed immunohistochemically. The superoxide dismutase (SOD), glutathione peroxidase (GSH‐Px) and malondialdehyde (MDA) activity levels in the testis tissue were also measured. The superoxide dismutase activity and malondialdehyde levels in the torsion group were significantly higher than those of the sham group (P < 0.05). Thymoquinone administration significantly reduced these levels. Torsion significantly increased active‐Caspase 3 and Bax expression, which was decreased by thymoquinone. The apoptotic index of the torsion group was significantly higher than that of the control group. However, thymoquinone significantly reduced the apoptotic index (P < 0.05). Our results indicate that thymoquinone plays a protective role in oxidative stress induced ischaemia–reperfusion in the testis tissue of rats.     

Anti‐oxidative Status and Semen Quality during Cooled Storage in Stallions

Activity of the anti‐oxidative enzymes glutathione peroxidase (GSH‐Px), superoxide dismutase (SOD) and catalase (CAT), content of thiobarbituric acid reactive substances (TBARS) and SH‐groups were determined in native stallion semen (n = 8 stallions). Semen was then diluted in Kenney extender, EquiPro^® extender either with or without addition of N‐acetyl cysteine or phosphate‐buffered saline (PBS) and stored for 72 h at 5°C. Correlations between initial activity of enzymes and development of semen motility and membrane integrity were calculated. Activities of GSH‐Px, SOD and CAT immediately after semen collections were 10.0 ± 0.6 picokatals, 0.40 ± 0.03 SOD units and 0.70 ± 0.05 nanokatals/10⁶ spermatozoa respectively. TBARS content was 0.06 ± 0.01 nmol and SH‐group content 1.7 ± 0.5 mmol/10⁶ spermatozoa. The loss of motile spermatozoa during storage did not differ between extenders. N‐acetyl cysteine had no effect on semen motility and membrane integrity. The loss in membrane‐intact spermatozoa was highest (P < 0.05) in semen diluted in PBS. Motility and membrane integrity after addition of extender were positively correlated with GSH‐Px and CAT, indicating that anti‐oxidative mechanisms contribute to the initial high percentage of motile and membrane‐intact spermatozoa. However, in these samples the decrease in semen quality was most pronounced. No correlations existed between initial activity of anti‐oxidative enzymes, peroxidation products and semen quality during storage. This indicates that once extender has been added, peroxidative damage to sperm membranes is not the predominant cause of losses in semen quality.     

Effect of Short‐term Hypothermia on Lipid Peroxidation and Antioxidant Enzyme Activity in Rats

This experiment was carried out to determine the effect of short‐term hypothermia on blood malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), glutathione peroxidase (GSH‐Px) and glucose‐6‐phosphate dehydrogenase (G‐6‐PD) concentrations in rats. Twenty Sprague–Dawley rats were used weighing 180–200 g and on average 3.5 months old. They were randomly divided into two experimental groups: control (without cooling) and hypothermic (with cooling). The rats of the hypothermic group were cooled by immersion into cold water (10–12 °C), and the control rats were immersed into water of body temperature (37 °C) up to the neck without using any anaesthetic or tranquilizer for 3 min Rectal body temperatures of both groups were measured and blood samples to analyse MDA, GSH, SOD, GSH, GSH‐Px and G‐6‐PD were collected immediately after the treatment. It was found that the MDA level was higher and the GSH and G‐6‐PD levels were lower in the hypothermic group than those in the controls. There was no difference between the control or hypothermic group regarding SOD or GSH‐Px levels. It is concluded that acute hypothermia increased the lipid peroxidation and decreased the GSH and G‐6‐PD levels in rats.     

Glutathione depletion and increased apoptosis rate in human cystinotic proximal tubular cells

We have determined levels of glutathione (GSH), ATP, mitochondrial complex activity and apoptosis rate in proximal tubular cells (PTCs) exfoliated from urine in cystinotic (n=9) and control (n=9) children. Intracellular GSH was significantly depleted in cystinotic PTCs compared with controls (6.8 nmol GSH/mg protein vs 11.8 nmol GSH/mg protein; P<0.001), but there were no significant differences in mitochondrial complex activities or ATP levels under basal conditions. Cystinotic PTCs showed significantly increased apoptosis rate. After PTCs had been stressed by hypoxia, there was further depletion of GSH in cystinotic and control PTCs (2.4 nmol GSH/mg protein vs 7.2 nmol GSH/mg protein; P<0.001). Hypoxic stress led to increased complex I and complex IV activities in control but not in cystinotic PTCs. ATP levels were significantly reduced in cystinotic PTCs after hypoxic stress (12.2 nmol/mg protein vs 26.9 nmol/mg protein; P<0.001). GSH depletion occurs in this in vitro model of cystinotic PTCs, is exaggerated by hypoxic stress and may contribute to reduced ATP and failure to increase complex I/IV activities. Apoptotic rate is also increased, and these mechanisms may contribute to cellular dysfunction in cultured, human cystinotic PTCs.     

Changes in the Level of Endogenous Leptin,FSH, 17β‐Oestradiol and Metabolites during Lupin‐induced Increase in Ovulation Rate in Ewes

The aim of the study was to determine the changes in the plasma concentration of leptin during lupin feeding‐induced increase in the ovulation rate (OR) in ewes. Additionally, alterations in the plasma level of glycogenic amino acids and glucose (as the factors influencing leptin secretion) and the levels of follicle‐stimulating hormone (FSH) and 17β‐oestradiol (E‐2) (as the hormones regulated by leptin and engaged in recruitment, selection and development of ovulatory follicles) were analysed. Ninety‐six female Polish Lowland Sheep were used. All ewes were cyclic and synchronized with PGF_2α. The ewes were divided into two groups: control (n = 48), fed only with hay, and experimental (n = 48), received additionally lupin (Lupinus angustifolius) grain as a high‐protein and a high‐energy supplement. They were given lupin from the second to 13th day of the oestrous cycle at increasing doses (150–750 g/day per ewe). On the 11th day of cycle blood samples for analysis of hormones, amino acids and total glucose concentration, were collected from the jugular vein. OR was determined by laparoscopy of ovaries on the sixth day of the following oestrous cycle. Mean OR of ewes supplemented with lupin grain (1.687 ± 0.463) was 30.67% higher than that of control (1.291 ± 0.454). In spite of the unchanged body mass, a significant increase (P ≤ 0.05) in mean concentration of plasma leptin in the experimental ewes [2.17 ± 0.15 ng/ml human equivalent (HE)] was found in comparison with control (1.42 ± 0.12 ng/ml HE). A significantly (P ≤ 0.05) higher plasma FSH level in the ewes fed lupin (105.21 ± 5.87 ng/ml) compared with those fed hay (67.88 ± 6.03 ng/ml) was also found. However, plasma level of E‐2 decreased after lupin feeding. Moreover, in the ewes fed lupin the plasma concentrations of glucose and nine glycogenic amino acids (Gly, Ala, Val, Met, Leu, Ile, Tyr, Phe and Arg) were increased. It can be concluded that lupin feeding exerts the stimulatory effect on the OR in Polish Lowland Sheep. The increase in OR is connected with significantly higher plasma leptin level and coincident with rise in FSH, glycogenic amino acids and glucose concentration. In contrast, the level of plasma E‐2 was significantly decreased in lupin‐fed ewes.     

Oxidative stress, inflammation and early cardiovascular damage in children with chronic renal failure

The relationship between inflammation, oxidant stress and cardiovascular damage in children with chronic renal failure (CRF) has not previously been investigated. The aim of this study was to investigate markers of oxidative stress, inflammation and early cardiovascular abnormalities. Therefore, erythrocyte superoxide dismutase (SOD) and catalase (CAT) activities; blood glutathione (GSH) and serum malondialdehyde (MDA) levels; C-reactive protein (CRP) and proinflammatory cytokines (IL-6, TNF-α,); and left ventricular masses (LVM) and intima media thicknesses (IMT) were measured in children with CRF. A total of 29 children with CRF (19 nondialysis, 10 peritoneal dialysis) were included. The control group consisted of 25 healthy subjects. CRF children had significantly increased IL-6, TNF-α, CRP and MDA concentrations and decreased SOD, CAT and GSH levels compared with controls (P<0.05). Nondialysis and peritoneal dialysis subgroups had similar oxidative stress and inflammation biomarkers (P>0.05). Erythrocyte CAT was positively correlated with CRP, TNF-α, and IL2-R in the study group. Positive correlations were found between cytokine concentrations, CRP and urea/creatinine levels. Significantly increased LVM and IMT values were found in CRF children (P<0.05). In conclusion, increased oxidant stress and inflammation together with early cardiovascular damage were found in CRF children. Further studies with more patients are needed to verify these results.     

The effects of chronic exposure to ethanol and cigarette smoke on the level of reduced glutathione and malondialdehyde in rat kidney

The aim of this study was to investigate the effects of chronic ethanol intake and cigarette smoke exposure on rat kidney. The animals were divided into four experimental groups: (1) the control group (C), (2) the ethanol group (E), (3) the cigarette smoke group (CS), and (4) the cigarette smoke plus ethanol group (CS+E). Apart from the control group, these were treated with ethanol and/or cigarette smoke for 6 months. The animals were killed and the kidneys removed to determine the levels of reduced glutathione (GSH) and malondialdehyde (MDA) and for histopathological analysis. The levels of GSH/g wet tissue were 1.58±0.09 µmol, 0.91±0.05 µmol, 1.14±0.06 µmol, and 0.82±0.04 µmol for C, E, CS, and CS+E, respectively. In groups of E, CS, and CS+E, the GSH values were significantly lower than that of group C animals (P<0.05). Although, we detected lower GSH levels in the CS+E than the CS group (P<0.05), a significant difference in GSH levels between CS+E and E was not observed. The levels of MDA/g wet tissue were 40.1±3.4 nmol, 71.4±2.8 nmol, 64.0±3.6 nmol, and 76.5±4.3 nmol, for C, E, CS and CS+E, respectively. In E, CS, and CS+E, the MDA values were significantly higher than in group C (P<0.05). The increase in MDA levels in CS+E were not significantly different from groups E or CS. Histopathological analysis of the kidney slices showed severe degeneration of the tissues. Advanced hydropic degeneration of kidney tubules was clearly observed in the CS group. In group E, advanced tubular and interstitial damage, mononuclear cell infiltration and tubular thyroidization were clearly visible. In group CS+E, an intense inflammatory cell infiltration was detected under the transitional epithelium. We conclude that chronic exposure to ethanol and cigarette smoke may cause an oxidative burst in rat kidney by increasing the formation of reactive oxygen species.     

Preventive Treatment with Vitamin E Alleviates the Poisoning Effects of Carbon Tetrachloride in Cattle

Fifteen yearling steers were used to study the preventive effect of vitamin E on the protection against free radicals produced by carbon tetrachloride (CCl₄). The animals were randomly divided in three equal groups and treated as follows: group A – previously injected (i.m.) with 15 IU/kg BW on the 15th and second day before the trial and drenched with 0.05 ml/kg BW CCl₄; group B – only drenched with the same dose of CCl₄; group C – drenched with a placebo. Food intake was recorded and blood samples collected daily for 8 days after the CCl₄ drenching to compare the activity of aspartate aminotransferase (AST), gamma‐glutamyltransferase (γ‐GT) and the levels of erythrocyte reduced glutathione (GSH) and serum malonyldialdehyde (MDA). Food intake was reduced in group B for the first 3 days (P < 0.05); higher activities of AST and γ‐GT were observed in the poisoned groups, nevertheless the overall values were lower in the group A than B (P < 0.02); only the group A reached the basal values of AST at the seventh day; higher levels of GSH and MDA were recorded in the poisoned cattle indicating the generation of free radicals. It was concluded that the preventive use of Vitamin E lessened the damage in hepatic tissue caused by the free radicals and prevented the anorexia caused by CCl₄.     

Fasting‐induced Changes in Ovulation Rate,Plasma Leptin,Gonadotropins, GH,IGF‐I and Insulin Concentrations During Oestrus in Ewes

The aim of this experiment was to study the changes in the hormonal status and ovulation rate (OR) evoked by starvation during the follicular phase of the oestrous cycle in ewes. To achieve this goal, 12 female crossbreed sheep were synchronized and then half of them were fasted from the 12th to the 16th day of the oestrous cycle. On the 16th day, analysis of hormones and insulin‐like growth factor‐I (IGF‐I) were performed in 10‐min intervals. Then, on the 6th day of the following oestrous cycle, the OR in all ewes was determined by laparoscopy. Fasting reduced significantly (P < 0.05) the OR in ewes (1.25 ± 0.50) in comparison with control (1.75 ± 0.50). The drop in the OR was coincident with a significant (P < 0.001) decrease in the plasma concentration and pulse amplitude of leptin (0.29 ± 0.08 ng/ml versus control 0.53 ± 0.14 ng/ml), the plasma level of luteinizing hormone (LH) (0.19 ± 0.06 IU/l versus 0.25 ± 0.09 IU/l in control; P < 0.05) and the mean frequency of LH pulses (2.0/h versus 2.5/h in control). Fasting resulted also in a significant (P < 0.05) decrease in the plasma concentration and pulse amplitude of follicle stimulating hormone (FSH) in comparison with the control. Simultaneously, a significant (P < 0.001) drop in the IGF‐I concentration in the fasted ewes (4.78 ± 0.91 ng/ml) was found in comparison with control (7.63 ± 1.85 ng/ml). Also the level of insulin were significantly (P < 0.001) lower in the fasted (178.99 ± 39.08 pm /l respectively) than in the control sheep (302.66 ± 49.01 pm /l respectively). Meanwhile, a double increase in the growth hormone (GH) pulses frequency and an augmentation in its plasma concentrations as a result of starvation was found. The obtained results shows that the acute fasting exerts an inhibitory effect on the ovulation rate in ewes coincident with suppression in leptin, FSH and LH secretion and changes in signalization mediated by GH.     

Berberine ameliorates experimental varicocele‐induced damages at testis and sperm levels; evidences for oxidative stress and inflammation

The present study was performed to show the ameliorative effect of berberine (BBR), as an antioxidant and anti‐inflammatory agent, against experimental varicocele (VCL)‐induced molecular and histological damages. For this purpose, 50 mature Wistar rats were divided into control, control‐sham, VCL‐sole, 50 mg/kg and 100 mg/kg BBR‐treated VCL‐induced groups. The tissue levels of interleukin‐6 (IL‐6), tumour necrosis factor‐α (TNF‐α), nitric oxide (NO), total antioxidant capacity (TAC), malondialdehyde (MDA), superoxide dismutase (SOD) and gluthatione peroxidase (GSH‐px) as well as the mRNA levels of testicular CuZn SOD, MnSOD, EC‐SOD and GSH‐px were evaluated. The serum concentration of testosterone and germ cells mRNA damage were analysed. Finally, the sperm viability, motility, DNA integrity and chromatin condensation were analysed. Observations revealed that, the BBR significantly downregulated VCL‐increased IL‐6, TNF‐α and NO levels, upregulated the CuZn SOD, MnSOD, EC‐SOD and GSH‐px mRNA level, decreased testicular MDA content, enhanced serum testosterone level and ameliorated testicular TAC, SOD and GSH‐px levels. The animals in BBR‐treated groups exhibited diminished mRNA damage versus non‐treated VCL‐induced group. The BBR has significantly (p < 0.05) improved sperm parameters. In conclusion, the BBR by promoting testicular antioxidant potential and by downregulating inflammatory reactions fairly promotes spermatogenesis and upregulates the sperm quality.     

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.     

The Possible Protective effect of L‐carnitine on Tilmicosin‐induced Cardiotoxicity in Mice

The protective effect of l ‐carnitine was investigated against tilmicosin‐induced cardiotoxic effects including blood creatine kinase (CK), CK‐MB, total sialic acid as well as the alterations in glutathione and malondialdehyde concentrations in mice. Thirty‐two Balb/C mice were divided into four groups including group 1 (control), group 2 (l ‐carnitine, s.c., 500 mg/kg for 5 days), group 3 (tilmicosin, s.c., single dose of 75 mg/kg) and group 4 (l ‐carnitine plus tilmicosin). Serum CK, CK‐MB and malondialdehyde (MDA) levels were significantly (P < 0.05) higher in group 3 compared with those of other groups. Total sialic acid level in group 3 was found to be significantly (P < 0.05) higher than that in groups 1 and 2, as well. Contrary to these results, glutathione level in group 3 was found to be significantly (P < 0.05) lower than that in groups 1 and 2. In group 4, serum CK, CK‐MB, MDA and total sialic acid levels were found to be significantly (P < 0.05) lower than those in group 3. These results suggest that tilmicosin is cardiotoxic in mice as evidenced by higher total sialic acid, CK and CK‐MB. In addition, tilmicosin caused the decrease in glutathione and increase in MDA levels. However, administration of l ‐carnitine could ameliorate these adverse toxic effects of tilmicosin in mice.     

Effects of quercetin and fish n‐3 fatty acids on testicular injury induced by ethanol in rats

The aim of this study was to investigate the effects of quercetin and fish n‐3 fatty acids on the changes in testis induced by ethanol. Forty‐five rats divided into five groups, control, ethanol, ethanol+quercetin, ethanol+fish n‐3 fatty acids and ethanol+quercetin+fish n‐3 fatty acids. At the end of 8 weeks, all the rats were sacrificed. Degenerative changes in histopathological analyses, the decreased body weight gain and seminiferous tubule diameters in ethanol group have been observed. TUNEL assay also showed an increase in apoptotic cell number. The activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH‐Px), xanthine oxidase (XO) and testosterone levels were decreased as well as the levels of malondialdehyde (MDA) and nitric oxide (NO) were increased in ethanol group. Histopathological changes caused by ethanol have been improved by quercetin and fish n‐3 fatty acids. It was also found that protection was provided by increasing SOD, CAT and GSH‐Px activities in groups administered quercetin, fish n‐3 fatty acids and quercetin+fish n‐3 fatty acids, and by decreasing the levels of MDA and NO in groups administered both quercetin and fish n‐3 fatty acids together. These results suggest that quercetin and fish n‐3 fatty acids are beneficial agents to reduce testicular injury induced by ethanol except for testosterone levels.     

Protective role of methylprednisolone and heparin in ischaemic‐reperfusion injury of the rat testicle

This study evaluated the therapeutic efficacy of heparin and methylprednisolone in the treatment of ischaemic reperfusion (IR) injury of the testis. Twenty‐four male Sprague‐Dawley rats were allocated equally into three groups of eight animals each. The left testes were rotated 720° for 2 h in the rats in the torsion–detorsion group. Rats in the treatment groups underwent the same surgical procedure as the torsion–detorsion group but were also given methylprednisolone (group II) or heparin (group III) by an intraperitoneal route 30 min prior to detorsion. Left orchiectomy was performed in all rats from each experimental animal at 2 h after detorsion, and the tissue was harvested for the measurement of malondialdehyde (MDA), protein carbonyl (PC) and nitric oxide (NO) and the endogenous antioxidant enzymes, superoxide dismutase (SOD), glutathione peroxidase (GSH‐Px) and catalase. Additional tissue was evaluated using histopathological and immunohistochemical changes. PC and MDA levels were significantly reduced in the treated groups compared to the control group. There was no statistically significant difference in NO level or SOD, GSH‐Px and catalase activity among the treatment groups. Histopathological and immunohistochemical findings supported biochemical changes. It is concluded that pre‐treatment with methylprednisolone or heparin protects the testis in ischaemic reperfusion injury caused by testicular torsion–detorsion.     

Effects on Skeletal Muscle Glutathione Status of Ischemia and Reperfusion Following Abdominal Aortic Aneurysm Surgery

Glutathione (GSH) is an important endogenous scavenger against reactive oxygen species. Elective abdominal surgery without ischemia and reperfusion leads to decreased muscle GSH concentrations 4-72 hr postoperatively without altering GSH redox status. In the present study, we investigated to what extent muscle GSH status was affected during and following elective abdominal aortic aneurysm repair. From patients (n = 10) undergoing abdominal aortic repair, thigh muscle specimens were taken preoperatively, at maximal ischemia, and at 10 min and 4, 24, and 48 hr of reperfusion. Specimens were analyzed for GSH, amino acids, and energy-rich compounds. At maximal ischemia, phosphocreatine decreased by 37% (p < 0.05) and lactate and creatine increased by 274% and 57% (p < 0.001 and 0.05), respectively, indicating ischemia during the clamping of aorta. Adenosine triphosphate, on the other hand, remained unaltered during the entire study period. Total GSH (tGSH) decreased by 46% at 24 hr and by 43% at 48 hr of reperfusion (p < 0.001), while reduced GSH decreased by 48% at 24 hr and by 44% at 48 hr (p < 0.001). The redox status (GSH/tGSH) of GSH and oxidized GSH remained unaltered. Among the constituent amino acids of GSH, glycine and cysteine remained unaltered while glutamine and glutamate decreased by 55% and 55%, respectively (p < 0.001). Abdominal aortic aneurysm repair induces metabolic alterations characteristic for ischemia. The antioxidative capacity in terms of muscle levels of GSH was decreased. However, the oxidative stress during reperfusion did not change GSH status more than what has been reported following abdominal surgery without ischemia and reperfusion. The results indicate that the oxidative stress elicited by elective abdominal aortic aneurysm repair is outbalanced by a compensated GSH metabolism not giving rise to an increased amount of oxidized GSH or an altered GSH redox status.     

The protective role of melatonin and curcumin in the testis of young and aged rats

We aimed to investigate the effect of melatonin and curcumin treatment on oxidative stress, apoptosis, and histology of testicular tissue in our study. Four groups were formed using young (4 months old, n = 6) and aged (20–22 months old, n = 18) male Wistar albino rats: (a) Young control (1% ethanol:phosphate‐buffered saline [PBS], subcutaneously [s.c.]); (b) Aged control (CTL; n = 6, 1% ethanol:PBS, s.c.); (c) Aged Melatonin (MLT; n = 6, 10 mg/kg, s.c.); (d) Aged Curcumin (CUR; n = 6, 30 mg/kg, i.p.). At the end of 21 days, the rats were sacrificed, and testicular tissues were removed. Malondialdehyde (MDA) in the testicular tissue was determined with thiobarbituric acid reactive substances formation, and glutathione (GSH) was determined with modified Ellman method; testosterone level was determined with chemiluminescence method and histologic changes were determined with Haematoxylin‐Eosin and Johnsen's scoring; Apoptotic cell counts were made with TUNEL staining of seminiferous tubule in testis. With ageing, MDA level increased in testicular tissue, but GSH and blood testosterone levels decreased. Melatonin treatment for aged rats significantly decreased Paired total testicular/body weight ratio compared to aged control group (p < 0.05). Curcumin treatment for aged rats significantly increased GSH level compared to the aged control group (p < 0.05). Besides, melatonin and curcumin treatment significantly decreased the number of apoptotic cells and significantly increased Johnsen's score (p < 0.05).