The Role of Reduced Glutathione on the Activity of Adenosine Deaminase, Antioxidative System, and Aluminum and Zinc Levels in Experimental Aluminum Toxicity.

Aluminum (Al) is one of the most abundant element in the world. But aluminum exposure and accumulation causes serious diseases, related with free radicals. Reduced glutathione (GSH) is a tripeptide with intracellular antioxidant effects. This study aimed to investigate the role of GSH on adenosine deaminase (ADA), antioxidant system, and aluminum and zinc (Zn) levels in acute aluminum toxicity. In this study, Sprague-Dawley rats (n = 32) were used. The rats were divided into four equal groups (n = 8). Group I received 0.5 mL intraperitoneal injection of 0.9% saline solution (NaCI), Group II received single-dose AlCI3, Group III was given GSH for seven days, and Group IV was given AlCI3 single dose, and at the same time, 100 mg/kg GSH was given for seven days. At the end of the trial, blood samples were collected by cardiac puncture. Serum total antioxidant status (TAS) and Zn levels were lower in the aluminum-administered group than the control group. In contrast, plasma total oxidant status (TOS) and aluminum concentrations and ADA activity were found higher in the aluminum-administered group than in the control group. Unlike the other groups, group GSH administrated with aluminum was similar to the control group. As a result, GSH administration has a regulatory effect on ADA activity, antioxidant system, and Zn levels in experimental aluminum toxicity. In addition, GSH may reduce the oxidant capacity increased by Al administration and may have a tolerant role on the accumulated serum Al levels. But long-term experimental Al toxicity studies are needed to reach a firm conclusion.

Oxidative stress in neonatal lambs and its relation to health status and passive colostral immunity.

Oxidative stress (OS) related to birth in ruminants may increase susceptibility to diseases. However, information about the physiological changes related to OS in lambs is limited. This study was designed to evaluate changes in OS parameters [total antioxidant status (TAS, mmol Trolox Equiv./L), total oxidant status (TOS, µmol H2O2 Equiv./L)] and calculated oxidative stress index (OSi = TOS/TAS * 100, Arbitrary Unit) and its association with passive colostral transfer immunity and health status in neonatal Akkaraman crossbreed lambs. Control group of healthy lambs (n = 15) were blood sampled before (day 0) and after colostrum intake (on day 1, 2, 4, 7, 14 and 28) in order to evaluate the effect of colostrum intake on TAS, TOS and OSi over time. In addition, a group of lambs (experimental group, n = 60), based on their health status and different cut off value of IgG (mg/dL), was also blood sampled at 24 ± 1 h of colostrum ingestion to determine the relationship between OS parameters, IgG and health status. OS was evident at birth and gradually decreased in control group as TOS (days 0:8.06 ± 0.64, 1:7.05 ± 0.51, 2:5.23 ± 0.35, 4:5.05 ± 0.2, 7:5.13 ± 0.34, 14:3.75 ± 0.12 and 28:3.73 ± 0.15 µmol H2O2 Equiv./L), OSi (days 0: 8.35 ± 0.64, 1: 9.17 ± 1.05, 2:6.83 ± 0.6, 4:6.30 ± 0.20, 7:6.67 ± 0.59, 14:4.46 ± 0.19 and 28:4.87 ± 0.29 au) and TAS (days 0: 0.97 ± 0.02, 1:0.82 ± 0.04, 2:0.81 ± 0.05, 4:0.81 ± 0.02, 7:0.80 ± 0.03, 14:0.85 ± 0.02 and 28:0.79 ± 0.03 mmol Trolox Equiv./L) values changed over time. In the experimental group, the lambs died in the first week of the neonatal period had higher levels of TOS (10.3 ± 0.90 µmol H2O2 Equiv./L) and OSi (1.61 ± 0.22 au) than healthy ones (TOS:6.90 ± 0.36 µmol H2O2 Equiv./L, OSi:1.12 ± 0.11 au) (P < 0.001). The gradual decrease in OS in the neonatal period was accepted as a sign of improved antioxidant response. The study disclosed that deaths may develop when OS at birth is not balanced by sufficient amount and quality of colostrum an important source of antioxidants such as enzymes and vitamins. The study presented may provide new information in the field of study on oxidative stress in neonatal lambs and help to prevent deaths in the first week of life. It can also be useful in developing preventive measures such as supplements of antioxidants to colostrum.

Hemoglobin-Inorganic Hybrid Nanoflowers with Different Metal Ions as Potential Oxygen Carrying Systems.

Protein-inorganic hybrid nanoflowers have tremendous potential in bionanotechnology due to their simple method of preparation, high stability and superior properties. Considering these features, the present study was designed to investigate the artificial blood substitution potentials of hemoglobin-inorganic hybrid nanoflowers. In this context, hemoglobin-inorganic hybrid nanoflowers (Cu-NF, Co-NF and Zn-NF) were synthesized using with different metal ions (copper, cobalt and zinc), then their oxygen carrying capacity, the hemolytic studies, in vitro oxidant/antioxidant capacity levels and oxidative stress index were reported for the first time. The present findings have revealed that Zn-NF had significant oxygen content and artificial oxygen carriers (AOC), as well as a significantly low percent hemolysis rate and a safe standard value. Also, hemolysis rate decreased along with the increases in hemoglobin content coupled with increments in nanoflower concentrations. The percentage hemolysis rate was lower than all nanoflowers at low free hemoglobin concentration, but hemolysis rates also increased with increments in concentration. The results showed that in general, Zn-NF stands out with its high total antioxidant capacity and low total oxidant capacity and oxidative stress index. The obtained results showed that Cu-NF and Co-NF, especially Zn-NF might be considered as a potential superior artificial oxygen carrier. Therefore, this nanoflower system might be act as an efficient material as a blood substitute in the near future.

Evaluating the activity and stability of sonochemically produced hemoglobin-copper hybrid nanoflowers against some metallic ions, organic solvents, and inhibitors.

The disadvantage of the conventional protein-inorganic hybrid nanoflower production method is the long incubation period of the synthesis method. This period is not suitable for practical industrial use. Herein, protein-inorganic hybrid nanoflowers were synthesized using hemoglobin and copper ion by fast sonication method for 10 min. The synthesized nanoflowers were characterized via scanning electron microscopy, energy-dispersive X-ray spectroscopy, X-ray diffraction, and Fouirer-transform infrared spectroscopy. The activity and stability of the nanoflowers in the presence of different metal ions, organic solvents, inhibitors, and storage conditions were also evaluated by comparing with free hemoglobin. According to obtained results, the optimum pH and temperatures of both hybrid nanoflower and free hemoglobin were pH 5 and 40 °C, respectively. At all pH levels, nanoflower was more stable than free protein and it was also more stable than the free hemoglobin at temperatures ranging between 50 °C and 80 °C. The free protein lost more than half of its activity in the presence of acetone, benzene, and N,N-dimethylformamide, while the hybrid nanoflower retained more than 70% of its activity for 2 h at 40 °C. The hybrid nanoflower activity was essentially increased in the presence of Ca2+, Zn2+, Fe2+, Cu2+ and Ni2+ (132%, 161%, 175%, 185% and 106%, respectively) at 5 mM concentration. The nanoflower retained more than 85% of its initial activity in the presence of all inhibitors. In addition, it retained all its activity for 3 days under different storage conditions, unlike free hemoglobin. The results demonstrated that new hybrid nanoflowers may be promising in different biotechnological applications such as catalytic biosensors and environmental or industrial catalytic processes.

Evaluation of organic-inorganic hybrid nanoflower's enzymatic activity in the presence of different metal ions and organic solvents.

In the present study, lipase-inorganic hybrid nanoflowers (L-hNFs) have indicated to possess several novel characteristics that are significant in terms of industrial worth. In detail, we reported the production of lipase-inorganic hybrid nanoflowers in different parameters and evaluated synthesized nanoflowers' enzymatic activity in the presence of different concentrations of metal ions and organic solvents. The optimum pHs of free enzyme, L-hNFs (0.02, 0.05, and 0.1) and L-hNFs (0.03) were pH 8, 7 and 6, respectively. L-hNFs (0.05) displayed the highest enzymatic activity in high alkaline pH values (10, 11, and 12) compared to both free enzyme and other L-hNFs. While the free enzyme showed no activity at pH 12, L-hNFs (0.05) maintained 57% of their activity. The optimum temperatures of the free lipase and L-hNFs were 30 °C and 40 °C, respectively. Free lipase activity decreased with temperature while L-hNFs had higher enzyme activity at 40 °C and 50 °C. Even in the case of increasing incubation and organic solvent and metallic ion concentration in which free lipase cannot completely maintain its activity, L-hNFs were able to protect their activity. L-hNFs exhibited excellent thermal and pH stability and high resistance to metallic ions and organic solvents at various concentrations for 3 h, 6 h, and overnight. This new lipase-inorganic hybrid nanobiocatalyst may be promising in many industrial processes such as chemical, biochemical, pharmaceutical, and biotechnological ones.

Kumiss Supplementation Reduces Oxidative Stress and Activates Sirtuin Deacetylases by Regulating Antioxidant System.

It was aimed to investigate the effects of kumiss a fermented mare horse beverage on the sirtuin deacetylases in the oxidative stress which had been induced by 1,2-dimethyl hydrazine (DMH). Forty BALB/C male mice were divided into four groups as control, kumiss (2 × 108 cfu/mL), DMH (20 mg/kg), and kumiss + DMH (2 × 108 cfu/mL + 20 mg/kg). At the end of 20-week regimen, SIRT2, SIRT3 protein expressions by western blotting, immunolocalizations, and inhibitory anti-oxidant activity analysis in liver, colon, and kidney tissues were performed. SIRT2 and SIRT3 expressions in DMH group were decreased in liver, colon, and kidney tissues and the decrease further stimulated by kumiss reinforcement. SIRT3, a mitochondrial protein, immunostaining increased in cell nuclei of tissues in response to kumiss treatments. The oxidative stress induced by DMH was determined to increase plasma 8-OH-2-deoxyguanosine, tissue oxidative stress index, and total oxidant capacity levels. Kumiss supplement was identified to reduce these levels and increase tissue total antioxidant capacity and reduced glutathione levels. Clarifying the molecular relationship between intracellular changes in the locations of SIRT2 and SIRT3 and oxidative stress might be important with regards to developing new medical treatments in the future. The kumiss may show a protective effect against DMH-induced damage by regulating the expression of sirtuin proteins and by protecting antioxidant system.

Boric acid and Borax Supplementation Reduces Weight Gain in Overweight Rats and Alter L-Carnitine and IGF-I Levels.

The aim of this study was to investigate the effects of boric acid (BA) and borax (BX) on live weight and obesity associated molecules including leptin, L-carnitine, insulin-like growth factor 1 (IGF-I), and heat shock proteins 70 (HSP70) in rats fed with high-fat diet. A total of 60 rats were equally allocated as ND (normal diet), HF (high-fat diet), HF+BA, HF+BX, ND+BX, ND+BA. Body weight increases in HF+BA (85 g) and HF+BX (86 g) were significantly lower (p<0.05) compared to HF group (126 g). Boron treatment decreased serum L-carnitine level in high-fat diet (HF+BA 11.12 mg/L, HF+BX 10.51 mg/L, p<0.05) compared to HF group (15.57 mg/L), while no change was observed in groups ND+BA (7.55 mg/L) and ND+BX (7.57 mg/L) compared to group ND (8.29 mg/L). Neither BA nor BX supplementation in ND and HF groups altered the serum levels of HSP70 and leptin. BA and BX supplementation in rats fed HF resulted in a significant reduction in live weight. Boron compounds altered L-carnitine and IGF-1 levels in rats. These results indicate that boron compounds are beneficial in the treatment of obesity as well as in the prevention of high-fat diet-induced weight increase. Alterations in serum L-carnitine and IGF-1 levels in boron treated rats also indicate possible role of boron compounds in energy metabolism in response to high fat diet.

The protective role of jervine against radiation-induced gastrointestinal toxicity.

In this study, we investigated whether jervine (J) could prevent gastrointestinal (GI) side effects of abdominopelvic radiotherapy (RT) in Wistar-Albino female rats. Rats were divided into five groups: control (C), J only (J), J administered at 5 mg/kg/days for 7 days, RT only (RT), J before RT (J + RT), J administered for seven days before RT, J both before and after RT (J + RT + J), and J administered for 7 days before RT and after RT for 3 days. The weights of rats were measured on the 1st, 7th, and 10th days of the study. Rats were sacrificed to obtain tissues from the liver and intestine, which was followed by taking blood samples intracardially. In addition, the tissues were stained with pyruvate dehydrogenase (PDH) immunohistochemically. In our study, J supplementation markedly reduced weight loss, and histopathological, immunohistochemical, biochemical results suggest that J had a protective effect on GI toxicity following RT.

Proteinase K hybrid nanoflowers (P-hNFs) as a novel nanobiocatalytic detergent additive.

In this study, enzyme-inorganic hybrid nanoflowers were synthesized using proteinase K and Cu2+ ions. The synthesized proteinase K-Cu2+ hybrid nanoflowers (P-hNFs) were characterized by their morphology and chemical point of view by using different techniques such as SEM, FTIR, EDX, and XRD. The proteolytic activities and some important characteristics such as optimum pH and temperature of the P-hNFs were also evaluated by comparison with free proteinase K. Optimum pH values of free proteinase K and P-hNFs were determined as pH 10 and pH 11, respectively. Optimum temperatures recorded for both free proteinase K (at pH 10) and P-hNFs (at pH 11) were 40 °C. In our study, for the first time, using some commercial detergents and surfactants, the utility of the P-hNFs as a detergent additive was also systematically evaluated. In these studies, the P-hNFs exhibited better activity than free proteinase K in the presence of all surfactants (CHAPS, DOC, SDS, Triton X-100 and Tergitol) except for Tween 80. Importantly, the P-hNFs was more stable and compatible with all tested solid laundry detergents. The findings demonstrated that the P-hNFs could potentially be used as an additive in detergent formulations.

A novel detergent additive: Organic solvent- and thermo-alkaline-stable recombinant subtilisin.

In the present work, subtilisin gene from Bacillus subtilis PTTC 1023 was synthesized, cloned into the vector pD441-NH and expressed in E. coli BL21 (DE3). Recombinant subtilisin was purified in a single-step procedure by affinity chromatography. The molecular mass of the purified protein was determined to be about 40kDa by SDS-PAGE. The optimum pH and temperature values of its proteolytic activity were 10.5 and 50°C, respectively and retained more than 70% and 89% of its activity in pH range of 7-12 and 30-60°C, respectively. Enzyme purity was estimated to be about 200- fold greater than that of the crude extract and subtilisin had a specific activity of 56.16U/mg, with a yield of about 87.9%. It was completely inhibited by phenylmethanesulfonyl fluoride, which strongly suggests its belonging to serine protease family. Interestingly, subtilisin protease displayed a significant compatibility with commercial detergents, and tolerance organic solvents, metallic ions and surfactants. The findings obtained demonstrated that protease of B. subtilis could potentially be used in future applications as an additive in detergent formulations.

The Therapeutic Role of Glutathione in Oxidative Stress and Oxidative DNA Damage Caused by Hexavalent Chromium.

Hexavalent chromium Cr (VI) causes various toxic and carcinogenic effects. The main carcinogenic effect is observed in the pulmonary system through inhalation route. Reduction of Cr (VI) to Cr (V, IV, and III) reactive intermediates within the cells by intracellular reducing agents such as glutathione is an important event leading to oxidative stress and oxidative DNA damage. This study evaluated the effects of intraperitoneal administration of Cr (VI) and GSH on total oxidant status (TOS), total antioxidant capacity (TAC), oxidative stress index, and oxidative DNA damage by evaluating the level of 8-hydroxy-2́-deoxyguanosine (8-OHdG) in Swiss-Albino mice. Seventy two mice were divided into 6 groups and treated intraperitoneally as follow: control (saline), group GSH (30 mg/kg GSH) groups of Cr-20 (20 mg/kg, K2Cr2O7), Cr-30 (30 mg/kg K2Cr2O7), Cr-20 + GSH (20 mg/kg K2Cr2O7 + 30 mg/kg GSH), Cr-30 + GSH (30 mg/kg K2Cr2O7 + 30 mg/kg GSH). Total oxidant capacities of Cr-20 and Cr-30 were increased compared to control, Cr-20 + GSH, and Cr-30 + GSH. TOS levels in Cr-20 + GSH and Cr-30 + GSH were lower than in Cr-20 and Cr-30. No difference in TAC was observed among the groups. 8-Hydroxy-2́-deoxyguanosine levels were increased in groups Cr-20 and Cr-30 compared with control and groups Cr-20 + GSH and Cr-30 + GSH. No difference was determined in 8-OHdG levels among control, groups GSH, Cr-20 + GSH and Cr-30 + GSH. Results indicate that Cr (VI) given i.p. route causes increased oxidative stress and oxidative DNA damage in the blood of Swiss-Albino mice. Administration of GSH via i.p. route protects from oxidative stress and DNA damage.

Histopathologic, biochemical and genotoxic investigations on chronic sodium nitrite toxicity in mice.

The aim of this study was to investigate the effects of long term Sodium nitrite (NaNO2) consumption. Swiss albino mice were given NaNO2 (0, 10 and 20 mg/kg/day) as mixed in feed for 8 months. At the end of treatments, animals were sacrificed and selected organs were processed for histopathologic, imunohistochemical, biochemical and genotoxic investigations. Mild to moderate degenerative changes were observed in liver, kidney, intestine, lung and spleen of NaNO2-given mice. Inducible nitric oxide synthase and nitrotyrosine activities increased in liver and kidney of NaNO2-given mice. Proliferating cell nuclear antigen activity increased in liver. Apoptotic cell death was observed in livers of the treatment groups. Liver malondialdehyde level was higher in the treatment groups while no change was seen in kidney. Nitric oxide levels in both liver and kidney of the treatment groups were lower than those of the control group. In genotoxic investigations, the number of chromosome and chromatid breaks, chromatid association, and polyploidy increased while mitotic index decreased in NaNO2-given mice. The results showed that NaNO2 would cause histopathologic changes, nitrosative tissue damage, and lipid peroxidation in liver and kidney, as well as induce chromosomal aberrations even if it was given at low levels for long time.

Subclinical mastitis causes alterations in nitric oxide, total oxidant and antioxidant capacity in cow milk.

The aim of this study was to investigate total antioxidant (TAC), and oxidant capacity (TOC) and nitric oxide (NO) levels in milk of cows with subclinical mastitis. Brown Swiss and Holstein breed cows were screened with California Mastitis Test (CMT) to determine mammary glands with subclinical mastitis. Moreover, somatic cell counts (SCC) were determined electronically in all milk samples. Mammary quarters were classified as healthy (n=25) or subclinical mastitis (n=35) based on CMT scores and somatic cell count (SCC: < or =200,000/ml or >200,000/ml) in milk. Nitric oxide, TOC and SCC levels were significantly higher (p<0.001, p<0.005 and p<0.001, respectively) in milk from mammary quarters with subclinical mastitis compared to those from healthy mammary quarters. In conclusion, subclinical mastitis results in higher NO concentrations, TOC and SCC, and NO and TOC were positively correlated with SCC. Moreover, alterations in NO levels and TOC in milk could be used as an alternative diagnostic tool to screen for subclinical mastitis.

Effects of dietary zinc and L-arginine supplementation on total antioxidants capacity, lipid peroxidation, nitric oxide, egg weight, and blood biochemical values in Japanese quails.

The aim of this study was to evaluate effects of dietary zinc and L-arginine supplementation on blood total antioxidant capacity (TAC), malondialdehyde (MDA), nitric oxide (NO), some blood chemistry parameters, and egg weights of laying quails. Three groups of Japanese quails were fed with a diet containing L-arginine (5 mg/kg), zinc (60 mg/kg), and normal basal diet (control) for 30 days. TAC, lipid peroxidation, and biochemical analysis were performed in the blood of animals. L-Arginine and zinc supplementation improved TAC and reduced MDA concentrations compared to the control (P<0.05). In comparison to the control, blood NO concentrations were increased by Larginine (P<0.01) and zinc treatment (P<0.05). Both zinc (P<0.001) and L-arginine (P< 0.01) supplementation significantly increased egg weight in laying quails. Some of the blood chemistry parameters were also altered by the treatment of L-arginine and zinc supplementation. No difference was found in blood albumin and creatinine levels among the groups. Blood glucose (P=0.833) and total protein (P=0.264) levels in control and Larginine-treated groups were found to be similar. Glucose and total protein levels were decreased in zinc-supplemented animals compared to the control and L-arginine groups (P< 0.05). No difference was found in triglyceride levels between control and zinc-applied groups (P=0.197). However, L-arginine treatment reduced the blood triglyceride levels compared to the control (P<0.05). In conclusion, L-arginine and zinc supplementation could be beneficial and effective for decreasing oxidative stress, boosting antioxidant capacity, and improving egg weight in the blood of the animals.

Hepatoprotective effect of L-carnitine against acute acetaminophen toxicity in mice.

L-carnitine is a cofactor in the transfer of long-chain fatty acid allowing the beta-oxidation of fatty acid in the mitochondria. It is also a known antioxidant with protective effects against lipid peroxidation. In this study, hepatoprotective effect of L-carnitine was investigated against acetaminophen (AA)-induced liver toxicity where mitochondrial dysfunction and oxidative stress are thought to be involved in AA hepatotoxicity. Sixty-four Balb/C mice were divided into eight groups. Mice were dosed with single-AA injection (500 mg/kg via the intra peritoneal route) with or without L-carnitine (500 mg/kg for 5 days starting 5 days before AA injection via intra peritoneal route) and sampled at 4, 8 and 24 h following AA injection. AA increased serum AST, ALT, total sialic acid (TSA) and MDA as well as tissue TSA and MDA levels significantly with the highest increase observed at 4 h, but there was a decrease in blood and tissue GSH level. Administration of L-carnitine significantly reduced AA-induced elevations in AST, ALT, TSA and MDA concentrations and increased GSH levels at all sampling points. AA also induced necrosis, hyperemia, sinusoidal congestion and hemorrhage with time-dependent increase in severity, but the degree of necrosis and histopathologic alterations were most severe at 24 h following AA administration. However, the degree of pathologic alterations was less severe with simultaneous L-carnitine application. These results suggest that AA results in oxidative damage in the liver with an acute effect. L-carnitine also has a prominent protective effect against AA toxicity and may be of therapeutic value in the treatment of AA-induced hepatotoxicity.