The effects of chronic smoking on lung tissue and the role of alpha lipoic acid

Abstract

We investigated the effects of alpha lipoic acid (ALA) on blood and lung tissue exposed chronically to cigarette smoke (CS). Female Sprague-Dawley rats were divided into three groups. Group 1 was the control group (CON): fresh air was supplied twice daily and 0.1 ml physiological saline was given orally for 8 weeks. Group 2 was exposed to CS: 12 cigarettes were smoked daily at two sessions for 1 h and 0.1 ml saline was given orally for 8 weeks. Group 3 (CS + ALA) was exposed to 12 cigarettes daily in two sessions for 1 h and 100 mg/kg/day ALA was given orally for 8 weeks. DNA damage was assessed using comet analysis; oxidative damage was assessed using ischemia-modified albumin (IMA) from blood; and total oxidant status (TOS), total antioxidant status (TAS) and oxidative stress index (OSI) were measured in blood and lung tissue. Histopathological and immunohistochemical evaluation of hypoxia-inducible factor (HIF)-1α, and ?2α, caspase-3, vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF₂) were conducted using lung tissue. The oxidative markers, TOS, OSI and IMA, and the comet analysis score were increased and the TAS level was decreased in the blood of the CS group compared to the CON group. IMA levels in blood, and TOS and OSI levels in the lung were decreased significantly in the CS + ALA group compared to the CS group. We observed increased septal wall thickness, marked and diffuse inflammatory reaction, emphysema, and necrotic cell debris in bronchial and bronchiolar lumens in the CS group. HIF-1α, HIF-2α, caspase-3 and FGF₂ expressions were increased, while VEGF expression decreased in the lung tissues of the CS group compared to the CON group. ALA slightly ameliorated the damage caused by chronic exposure to CS in the lungs, but further investigation is needed to determine its possible protective effects at different dosages.     

Effect of dried corn distillers’ grains with solubles and soybean meal supplements on physiological indicators and reproductive performance of ewes

This study aimed to evaluate the effects of adding dried distillers’ grains with solubles (DDGS) to the nutritional regimens of ewes at different time periods on reproductive traits, serum hormones and serum metabolites. In Experiment 1, 100 ewes were divided into 4 groups (n = 25) according to diet. The four diets were grass hay (H) alone (group H), H with barley (group H + B), H with DDGS for the whole feeding period (27 d) (group H + DDGS) or H and DDGS for 5 d (d 8–12 of oestrus cycle) (group H + DDGS5). Serum progesterone concentrations were affected by oestrus cycle (p < 0.001), but not by dietary treatments. However, feeding H + DDGS caused significant increases in serum insulin, leptin and growth hormone concentrations (p < 0.05). In Experiment 2, 30 ewes were divided into 2 groups (n = 15), receiving DDGS or soybean meal (SBM) during the prepartum period. Diets had no significant effect on weights of dams or lambs at birth; however, the weaning weights of lambs born from ewes of group DDGS were significantly higher (p < 0.05). Moreover, serum glucose, blood urea nitrogen, triglyceride and beta hydroxybutyric acid (BHBA) concentrations during the prepartum period were affected by dietary treatments and by time (p < 0.001). During the prepartum period, BHBA concentrations of ewes fed SBM were higher, while BHBA levels decreased during the last weeks of pregnancy regardless of diet. Lamb serum immunoglobulin G concentrations increased from 1 h to 24 h after birth (p < 0.001). Colostrum of ewes fed SBM had higher fat-free dry matter (DM) and protein contents in comparison to colostrum of ewes fed DDGS (p < 0.05). In conclusion, DDGS can be included as protein source in pregnancy rations up to 15% of DM to obtain reproductive performance outcomes equal to or exceeding those obtained with SBM.     

Synergic Effect of Exercise and Lipoic Acid on Protection Against Kainic Acid Induced Seizure Activity and Oxidative Stress in Mice

Anti-convulsant effects of physical exercise and lipoic acid (LA), also referred to as thioctic acid with antioxidant activity, were investigated using chemical induced seizure model. We investigated the synergic effect of physical exercise and LA on kainic acid-induced seizure activity caused by oxidative stress. After 8 weeks of swimming training, body weight decreased and endurance capacity increased significantly compared to sedentary mice. Kainic acid (30 mg/kg, i.p.) evoked seizure activity 5 min after injection, and seizure activity peaked approximately 80 min after kainic acid treatment. Median seizure activity score in KA only treated group was 4.55 (range 0.5–5), 3.45 for “LA + KA” group (range 0.5–4.3), 3.12 for “EX + KA” group (range 0.05–3.4, p < 0.05 vs. “KA only” group), 2.13 for “EX + LA + KA” group (range 0.5–3.0, p < 0.05 vs. “EX + KA” group). Also, there was a synergic cooperation of exercise and LA in lowering the mortality in kainic acid treated mice (χ² = 5.45, p = 0.031; “EX + KA” group vs. “LA + EX + KA” group). In addition, the synergic effect of exercise and LA was found in PGx activity compared to separated treatment (“LA + EX + KA”: 37.3 ± 1.36; p < 0.05 vs. “LA + KA” and “EX + KA” group). These results indicate that physical exercise along with LA could be a more efficient method for modulating seizure activity and oxidative stress.     

Effects of alpha-lipoic acid on high fructose induced hepatic pathology

Little is known about the pathogenesis of high fructose corn syrup (HFCS) induced hepatic toxicity. We investigated hepatic lesions induced by chronic HFCS consumption and the protective effects of alpha-lipoic acid (ALA) on liver pathology. We used 24 rats allocated randomly into three groups of eight. The HFCS group was given in drinking water for 10 weeks. The ALA + HFCS group was given the same dose of HFCS and ALA also was administered during the last 6 weeks of the experiment. The control group was untreated. The rats were euthanized at the end of 10 weeks and 24 h after the last ALA administration. A significant increase was observed in the serum aspartate aminotransferase (AST) level of the HFCS group compared to controls. Tissue malondialdehyde (MDA) levels also increased significantly and catalase (CAT) activity decreased significantly in the HFCS group. Caspase-3 expression increased significantly in the HFCS group compared to controls. In the ALA treated group, the levels of MDA, CAT and caspase-3 returned to near control levels. HFCS caused hepatic toxicity by increasing oxidative stress and apoptosis. ALA administration ameliorated the pathological changes.     

Effect of diabetes on collagen metabolism and hypoxia in human gingival tissue: a stereological,histopathological, and immunohistochemical study

Diabetes mellitus and periodontitis are chronic inflammatory diseases that disrupt soft tissue metabolism. The diseases separately or together increase apoptosis in gingival fibroblast cells and reduce cell renewal. We investigated the effects of diabetes and periodontitis on the composition and structure of gingival connective tissue. We used gingival biopsies from 16 healthy individuals (control group, C), 16 type 2 diabetic patients with chronic periodontitis (diabetes + periodontitis group, D + P) and 16 healthy chronic periodontitis patients (periodontitis group, P). Biopsies were obtained under local anesthesia. Clinical attachment level (CAL), gingival index (GI) and plaque index (PI) were measured prior to gingival biopsies. Fibroblast cells were counted stereologically. Inflammatory cells were counted histomorphometrically. Hypoxia-inducible factor (HIF)-1α, lysyl hydroxylase (PLOD-2), neutrophil collagenase (MMP-8), and vascular endothelial growth factor (VEGF) levels were evaluated immunohistochemically. CAL, GI and PI for the C group were lower than for the other groups (p < 0.05). Fibroblast cell counts were lower for the D + P group than for the other groups (p < 0.05). Diabetes increased inflammatory cell numbers in the D and D + P groups compared to the C and P groups. MMP-8 levels were higher for the D + P group than for the other groups. VEGF was elevated in both the P and D + P groups compared to the C group, while HIF-1α and PLOD-2 levels were comparable. Diabetes increased tissue destruction and inflammation, and decreased fibroblast cell numbers without affecting collagen crosslinking and HIF-1α levels.     

Effect of Different Parthenogenetic Activation Methods on the Developmental Competence of in vitro Matured Porcine Oocytes

The aim of this study was to investigate the effect of electrical pulse, ethanol, and ionomycin combined with cycloheximide (CHX), cytochalasin B (CB), and 6-dimethylaminopurine (6-DMAP) on parthenogenetic developmental competence of in vitro matured porcine oocytes. In experiment 1, oocytes were treated with direct current electrical pulse (DC pulse) and then incubated in the NCSU-23 medium supplemented with CHX, 6-DMAP, CB + CHX, and CB + 6-DMAP for 6 h, respectively. The rate of blastocyst development in DC pulse + CB + 6-DMAP group was significantly higher than those in other groups (42.4% vs 23.9% ～ 35.8%; P < 0.05); however, there were no differences in both of the cleavage rate and the cell number of blastocysts among four groups. In experiment 2, oocytes were treated with NCSU-23 medium containing 20 μM ionomycin for 40 min and then incubated in the NCSU-23 medium supplemented with CHX, 6-DMAP, CB + CHX and CB + 6-DMAP for 6 h, respectively. The rates of cleavage and blastocyst development in ionomycin + 6-DMAP group were higher than those obtained in other groups (66.2% vs 46.3% ～ 57.3%; 22.3% vs 7.4% ～ 16.1%; P < 0.05). In experiment 3, the activation effects of ethanol combined with 6-DMAP, CHX, CB + 6-DMAP and CB + CHX were investigated. The rates of cleavage and blastocyst development in ethanol + CB + 6-DMAP group were significantly higher than those in other groups (55.5% vs 42% ～ 46.2%; 18.0% vs 7.1% ～ 11.9%; P < 0.05). In experiment 4, the optimal activation protocols in each group plus DC pulse + ionomycin + 6-DMAP were compared. The results showed the rates of cleavage in DC pulse + CB + 6-DMAP group and ionomycin + 6-DMAP were higher than those in ethanol + CB + 6-DMAP and DC pulse + ionomycin + 6-DMAP (73.8–74.4% vs 56.5–57.5%; P < 0.05), but the blastocyst development only in DC pulse + CB + 6-DMAP group was significantly higher than that in other groups (34.1% vs 13.4% ～ 22.3%; P < 0.05). Total cell number of blastocysts in the group of DC pulse + ionomycin + 6-DMAP was higher than that in other groups (34.1 vs 25.3–27.2; P < 0.05). In conclusion, DC pulse, ethanol, CB, and 6-DMAP all affected the parthenogenesis of porcine oocytes matured in vitro, but their combination of DC pulse + CB + 6-DMAP showed the best result in both of cleavage and blastocyst development.     

Physiological Responses of Pinus sylvestris var. Mongolica Seedlings to the Interaction Between Suillus luteus and Trichoderma virens

The effects of the interaction between Suillus luteus (L.) Roussel and Trichoderma virens (J.H. Mill., Giddens & A.A. Foster) Arx on Pinus sylvestris var. mongolica Litv. were studied using plant physiology, mycorrhizal science, forest pathology, and biochemistry. Seedling growth and physiological parameters were determined, including the colonization rate of mycorrhizal fungi, biomass, root activity, photosynthetic pigment content, soluble protein content, antioxidant enzyme activities, rhizosphere soil enzyme activities, and protective enzyme activities. In addition, an optimal resistance system involving T. virens, mycorrhizal fungus (S. luteus), and P. sylvestris var. mongolica seedlings was constructed. Synergies between S. luteus and T. virens were observed, and most of the parameters of P. sylvestris var. mongolica seedlings inoculated with S. luteus 30 days + T. virens were higher than other treatments. After three months, when compared the control, the S. luteus 30 days + T. virens treatment gave increases in height (42.3 %); collar diameter (66.7 %); fresh weight (54 %); dry weight (50 %); soluble protein content (69.86 %); root activity (150 %); chlorophyll a (77.6 %); chlorophyll b (70.5 %); carotenoids (144 %); CAT activity (876.9 %); POD activity (268.3 %); SOD activity (66.18 %); β-1,3-glucanase activity (125.8 %); chitinase activity (40 %); rhizosphere soil catalase activity (97.8 %); and phosphatase activity (266.7 %). These results indicate that there may be a stimulating factor between S. luteus and T. virens when they are inoculated together (S. luteus 30 days + T. virens).     

Effects of 5-aminolevulinic acid on Swiss chard (Beta vulgaris L. subsp. cicla) seedling growth under saline conditions

Salinity is a widespread adverse environmental problem globally, and significantly limits crop production. In this study, the possibility of enhancing salinity stress tolerance of Swiss chard (Beta vulgaris L. var. cicla) by 5-aminolevulinic acid (ALA) foliar application was investigated. The Swiss chard plants were grown in hydroponic culture. Twelve-week-old uniform seedlings were treated by 0 and 40 mM saline regimes generated by the mixture of sodium chloride and sodium sulfate (molar ratio NaCl:Na₂SO₄ = 9:1), and were foliar-sprayed with 0 and 60 μM L^?1 ALA (every 3 days) for 6 days; then the plants were treated for another 7 days (every 3 day) with increased concentration of salinity and ALA, 80 mM and 120 μM L^?1. Salinity without ALA application significantly decreased plant growth [43 % in shoot dry weight (DW), 21 % in root DW, 24 % in relative growth rate (RGR), 43 % in leaf area (LA)], water uptake [20.8 % in relative water content (RWC), 47.9 % in osmotic potential (OP)], chlorophyll (Chl) a content (10 %), P_n (36 %), G_s (72 %) and T_r (59 %) compared with those in control plants; however, under saline conditions, ALA foliar application improved plant growth (49.7 % in shoot DW, 27 % in root DW, 42.3 % in RGR, 72.1 % in LA) and increased RWC (12 %), Chl a content (10 %) and photosynthetic parameters (27 % in P_n, 28 % in G_s, 14 % in T_r) compared with those in untreated plants. Salinity significantly increased Na⁺ content, resulting in the reduction of Mg²⁺ and K⁺ contents. ALA foliar application alleviated ionic toxicity through the reduction of Na⁺ content and Na⁺/K⁺ ratio. On the other hand, it increased total nitrogen and glycine betaine (GB) content. ALA foliar application slightly reduced malondialdehyde (MDA) content, indicating that ALA has the potential to alleviate oxidative stress in salinity-stressed Swiss chard.     

Macro- and Microelement Status in Animal and Human Hypertension: the Role of the ACE Gene I/D Polymorphism

Growth hormone (GH) and zinc (Zn) were evaluated for their potential to prevent radiation injury using a rat model of radiation-induced skin injury. Sprague–Dawley rats were divided into five groups: a control group not receiving Zn, GH, or irradiation: a radiation (RT) group receiving a single 30 Gy dose of gamma irradiation to the right hind legs; a radiation + GH group (RT + GH) receiving a single 30 Gy dose of gamma irradiation plus the subcutaneous administration of 0.01 IU kg d^?1 GH; a radiation + Zn group (RT + Zn) receiving a single 30 Gy dose plus 5 mg kg d^?1 Zn po; and a radiation + GH + Zn group (RT + GH + Zn) group receiving a single 30 Gy dose plus subcutaneous 0.01 IU kg d^?1 GH and 5 mg kg d^?1 Zn po. Acute skin reactions were assessed every 3 days by two radiation oncologists grouping. Light microscopic findings were assessed blindly by two pathologists. Groups receiving irradiation were associated with dermatitis as compared to the control group (P < 0.05). The severity of radiodermatitis in the RT + GH, RT + Zn, and RT + GH + Zn groups was significantly lower than that in the RT group (P < 0.05). Furthermore, radiodermatitis was observed earlier in the RT group than in the other treatment groups (P < 0.05). GH and Zn effectively prevented epidermal atrophy, dermal degeneration, and hair follicle atrophy. The highest level of protection against radiation dermatitis was observed in the combination group.     

Infradian rhythm in hematology,cell mediated immunity and serum micronutrient concentration around peripartum periods in high yielding crossbred cows before and after micronutrient supplementation

Alteration in hematology and cell-mediated immunity around peripartum periods before and after micronutrient supplementation was studied in this investigation. Twelve (12) high-yielding cross-bred cows in advanced pregnancy were selected for the experiment and divided into two equal groups viz. supplemented (n = 6) and unsupplemented (n = 6). Supplemented animals were provided with a micronutrient mixture (@ 25 g per cow) as per the recommendation of NRC over the normal feeding. Unsupplemented group of cows were kept as control with standard farm feeding management. Blood samples (15 ml/cow) were collected from all the animals during days 30th, 15th, and 7th before calving, on the day of calving, and during days 7th, 15th, and 30th after calving and analyzed for hematology viz. hemoglobin (Hb), total erythrocyte counts (TEC), total leukocyte counts, and differential leukocyte counts. In vitro phagocytic index (PI) of neutrophils and lymphocyte proliferation response (LPR) were also evaluated. Copper (Cu), zinc (Zn), manganese, and selenium (Se) concentrations in serum were estimated by atomic absorption spectrophotometer. The Hb concentration was found to be significantly (p < 0.01) higher in supplemented group compared to the unsupplemented group of cows. There were no significant differences in Hb concentration during different days of peripartum in both the group of cows. There was a significantly (p < 0.001) higher TEC in supplemented group compared to unsupplemented group of cows, though it did not vary significantly between different days of peripartum in both the group of cows. There was a significantly (p < 0.001) higher PI in supplemented group compared to the unsupplemented group of cows. In both the groups, the PI of blood neutrophils was significantly (p < 0.001) lower during calving. The LPR did not differ significantly between the groups, but it differed significantly (p < 0.01) between different days of peripartum and was lowest on the day of calving in both the groups. The Cu, Zn, and SE concentration was found to be significantly (p < 0.01) higher in supplemented group compared to the unsupplemented group of cows, but remained unaltered throughout the peripartum period. The study indicated that there was a marked immune depression around peripartum (15 days precalving to 15 days postcalving) in terms of in vitro phagocytic activity of blood neutrophils and in vitro LPR. Supplementation of micronutrients improves the hematological status as well as immune status, so it was recommended to supplement these micronutrients for better productivity and disease resistance in high-yielding cross-bred cows around peripartum.     

The role of cystatin C in vascular remodeling of balloon-injured abdominal aorta of rabbits

This study aimed to evaluate the role of cystatin C (CysC) in the vascular remodeling of balloon-injured abdominal aorta of rabbits. Forty-eight New Zealand white rabbits were randomly divided into three groups: the balloon-injured injury group (n = 16), the CysC monoclonal antibody group (n = 16), and the sham-operative group (n = 16). Serum CysC levels were detected by enzyme linked immunosorbent assay. Changes in adventitial area, adventitial thickness, lumen area (LA), neointimal area (IA), internal elastic lamina area (IELA), external elastic lamina area (EELA), vascular remodeling index (VRI) and residual stenosis (RS) were measured by the Leica image analysis system. Immunohistochemical analysis of α-smooth muscle actin (α-SMA) and proliferating cell nuclear antigen (PCNA) were performed. Serum CysC levels of rabbits in the balloon-injured injury group were significantly higher than those in the CysC monoclonal antibody group and the sham-operative group (both P < 0.05). At 6 weeks after balloon injury, the adventitial area and thickness, LA, IA, IELA and EELA in the balloon-injured injury group were also higher than those in the CysC monoclonal antibody and sham-operative groups (all P < 0.05). In addition, the balloon-injured injury group showed higher VRI and RS than those of the CysC monoclonal antibody group (both P < 0.05). The positive expression of α-SMA in the vascular adventitia and media in the balloon-injured group were higher than that of the CysC monoclonal antibody and sham-operative groups. The balloon-injured group also showed a stronger expression of α-SMA in the neointima than that of the CysC monoclonal antibody group. There was a strong positive expression of PCNA in the vascular adventitia and neointima in the balloon-injured and CysC monoclonal antibody groups. However, the number of PCNA-positive cells in the balloon-injured group was higher than that of the CysC monoclonal antibody group (25.45 ± 4.21 vs. 6.75 ± 1.11, P = 0.003). Our findings provide empirical evidence that serum CysC levels may play an important role in the vascular remodeling of balloon-injured abdominal aorta of rabbits.     

Effects of autocrine vascular endothelial growth factor (VEGF) in non-small cell lung cancer cell line A549

It is reported that the autocrine loop of the vascular endothelial growth factor (VEGF) is crucial for the survival and proliferation of non-small cell lung cancer (NSCLC) tumors. In this study we aimed to systematically investigate the role of autocrine vascular VEGF in NSCLC cell line A549 through inhibition of endogenous VEGF. A549 cells were transfected with florescence-labeled VEGF oligodeoxynucleotide with lipofectamine. For the experimental group, cells were transfected with VEGF anti-sense oligodeoxynucleotide (ASODN), sense oligodeoxynucleotide (SODN) and mutant oligodeoxynuleotide (MODN) respectively. For the control group cells were mock transfected with lipofectamine or culture medium. At indicated time point after transfection, the expression levels of VEGF mRNA and protein in A549 cells were analyzed by RT-PCR and ELISA respectively. Cell viability was measured by the MTT assay. Cell cycle distribution was detected by flow cytometry. As revealed by RT-PCR assay, the mRNA level of VEGF in cells transfected with ASDON was significantly lower than the other four groups (P < 0.05) at 24 and 48 h after transfection. ELISA assay yielded similar result with significantly decreased level of VEGF protein expression (P < 0.05). The survival fraction of A549 cells transfected with ASDON was significantly lower than the other four groups (P < 0.05) at 24 h after transfection. Also the percentage of G2 phase cells of ASODN group was significantly lower than other four groups. Our data indicate that VEGF expression is efficiently inhibited in A549 cells by ASODN transfection and this inhibition leads to inhibited cell growth and impaired cell cycle distribution.     

Synergistic Effects of Genistein and Zinc on Bone Metabolism and the Femoral Metaphyseal Histomorphology in the Ovariectomized Rats

Studies have shown that non-alcoholic fatty liver disease (NAFLD) patients are more prone to cardiovascular disease (CVD). Zinc and selenium deficiency are common in NAFLD. But the effects of zinc and selenium co-supplementation before and/or after disease progression on CVD markers are not clear in NAFLD patients. This study aimed to compare the effects of zinc and selenium co-supplementation before and/or after disease progression on some of the CVD markers in an experimental model of NAFLD. Forty male Sprague Dawley rats (197 ± 4 g) were randomly assigned into four dietary groups: control group (C; received 9% of calorie as fat), model group (M; received 82% of calorie as fat), and supplementation before (BS) or after (AS) disease progression. Animals were fed diets for 20 weeks in all groups. Fasting plasma glucose (FPG), insulin, HOMA-IR, ALT, AST, lipid profile, malondialdehyde (MDA) and vascular endothelial growth factor (VEGF) levels were measured as CVD indices. Serum ALT, AST, FPG, insulin, MDA, VEGF and HOMA-IR were significantly higher in the M than C group. Co-supplementation reduced serum ALT and AST levels in the BS and AS groups compared with the M group. FPG, insulin, HOMA-IR, VEGF, MDA, LDL/HDL-c and TC/HDL-c ratio were significantly reduced in the AS compared with the M group. TG/HDL-c ratio was significantly reduced in the BS and AS compared with the M group. Serum MDA, VEGF, Insulin and HOMA-IR were significantly lowered in the AS than BS group (p < 0.05). Zinc and selenium co-supplementation after NAFLD progression reduced CVD risk indices in an experimental model.     

Effect of Inducers on the Production of 5‐Aminolevulinic Acid by Recombinant Escherichia coli

Abstract

Aminolevulinic acid (ALA) was produced by recombinant Escherichia coli BL21(DE3) (pET28‐A.R‐hemA) harboring the ALA synthase gene (hemA) from Agrobacterium radiobacter zju‐0121. The effects of inducers on the ALA synthase activity and ALA productivity were evaluated. The results indicated that a low isopropyl‐β‐D‐thiogalactoside (IPTG) concentration (0.05 mmol/L) was favorable for high expression of ALA synthase, which resulted in higher ALA productivity. For metabolic engineering applications, lactose was a better substitute of IPTG for active enzyme expression. When lactose concentration was 5 mmol/L, the specific ALA synthase activity and ALA productivity reached 16.7 nmol/(min · mg of protein) and 1.15 g/L, respectively, which were about 15% and 43% higher than those induced by IPTG.     

Dynamic feeding low and high methionine diets affect the diurnal rhythm of amino acid transporters and clock related genes in jejunum of laying hens

This study was conducted to investigate effects of dynamic feeding low and high methionine (Met) diets on circadian clock gene expression in jejunum of laying hens. A total of 180 laying hens (Brown Hy-line, 41 weeks old) were allocated into 3 groups with 6 replicates each. The CON group received diet containing 0.30% Met, LH group received low Met diet (0.27% Met) at 07:30 h and high Met diet (0.33% Met) at 15:30 h, while HL group received high Met diet at 07:30 h and low Met diet at 15:30 h. After 10 weeks, jejunum samples were collected at 4 h intervals in a daily cycle initial starting at 07:30 h before feeding. Results showed that dynamic feeding of Met significantly affected the wave of mRNA expression of B0AT1, LAT1, SNAT2, CAT-2, and y⁺LAT2 by feeding regime during a day (P<0.05). Dynamic feeding of Met also influenced the fluctuation of relative mRNA expression of Clock, Bmal1, Cry1, Cry2, Per2, and Per3 significantly in hens during a day (P<0.05). These results demonstrated that dynamic feeding different Met level diets of hens affected the rhythmic fluctuation of amino acid transporters and may affected intestinal function by influencing the clock-related genes further.     

Regional Distribution of Longevity Population and Elements in Drinking Water in Jiangjin District,Chongqing City,China

This study was carried out to determine the protective effects of lithium borate (LTB) on blood parameters and histopathological findings in experimentally induced acute cadmium (Cd) toxicity in rats. Twenty-eight male Wistar albino rats were used, weighing 200–220 g, and they were randomly divided into four groups, including one control and the following three experimental groups: a Cd group (0.025 mmol/kg), a LTB group (15 mg/kg/day orally for 5 days), and a LTB + Cd group (15 mg/kg/day orally for 5 days and Cd 0.025 mmol/kg by intraperitoneal injection on the fifth day). All the rats in the study were anesthetized with ketamine at the end of the sixth day, blood was taken from their hearts, and then the rats were decapitated. The values in the control and LTB group were usually close to each other. White blood cell (WBC), neutrophil %, and C-reactive protein (CRP) levels increased in the Cd and LTB + Cd groups while lymphocyte and monocyte levels decreased in a statistically significant manner, in comparison to the other groups. It was determined that the levels of red blood cells (RBCs), hematocrit (Htc), and hemoglobin (Hb) did not change in the groups. The levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the Cd and LTB + Cd groups significantly increased, in comparison to the other groups, while the glucose, alkaline phosphatase (ALP), albumin (ALB), and total protein (TP) levels decreased. According to histopathological findings in the control and LTB groups, the liver and kidney tissues were found to have normal histological structures. In the Cd group, severe necrotic hemorrhagic hepatitis, mild steatosis, and mononuclear cell infiltration were detected in the liver. In the LTB + Cd group, degeneration and mild mononuclear cell infiltration were found in the liver. Regarding the kidney tissue in the Cd group, severe intertubular hyperemia in both kidney cortex and medulla, as well as degeneration and necrosis in the tubulus epithelium, was observed. In the LTB + Cd group, mild interstitial hyperemia and mononuclear cell infiltration was detected. Resultantly, it can be said that LTB at this dose has non-toxic effects and some beneficial effects for liver and kidney damage caused by acute Cd toxicity.     

Content of Minerals and Fatty Acids and Their Correlation with Phytochemical Compounds and Antioxidant Activity of Leguminous Seeds

This study was carried out to determine the effect of zinc on oxidative DNA damage in rats with experimental acute and chronic kidney deficiency. Six groups of five Wistar-Albino rats each were assigned as controls (C), acute kidney deficiency (AKD), zinc-supplemented (+Zn), acute kidney deficiency, zinc-supplemented (AKD + Zn), chronic kidney deficiency (CKD) and zinc-supplemented chronic kidney deficiency (CKD + Zn). The levels of 8-Oxo-2′-deoxyguanosine (8-OHdG) were determined, being the lowest in the CKD group (p < 0.05), higher in the C group than those of rats with CKD but lower than that of all the other groups (p < 0.05). There were no significant differences between the controls and the CKD + Zn group, or between the AKD and the +Zn groups. Among all groups, the highest 8-OHdG level was found in the AKD + Zn group (p < 0.05). DNA damage was greater in acute renal failure than in rats with chronic renal failure. The DNA damage in the zinc group was significantly higher than in the controls.     

Alpha Lipoic Acid Alleviates Oxidative Stress and Preserves Blood Brain Permeability in Rats with Subarachnoid Hemorrhage

The neuroprotective effect of alpha lipoic acid (ALA; 100 mg/kg, po), a dithiol antioxidant, on experimentally induced subarachnoid hemorrhage (SAH) was assessed in Wistar albino rats. Neurological examination scores recorded at the 48th h of SAH induction were increased in SAH groups, which were accompanied with significant increases in the formation of reactive oxygen species, DNA fragmentation ratios, malondialdehyde levels and myeloperoxidase activity, while significant decreases in the brain glutathione content and Na⁺, K⁺-ATPase activity were observed. On the other hand, ALA treatment reversed all these biochemical indices as well as SAH-induced histopathological alterations. Increased brain edema, impaired blood-brain-barrier permeability and neurological scores were also improved by ALA treatment. The results demonstrate that ALA exerts neuroprotective effects via the enhancement of endogenous antioxidant enzyme activity, the inhibition of neutrophil accumulation and free radical generation, suggesting a therapeutic potential in reducing secondary injury after SAH in patients.     

Augmentation of Fluvastatin Cytotoxicity Against Prostate Carcinoma PC3 Cell Line Utilizing Alpha Lipoic–Ellagic Acid Nanostructured Lipid Carrier Formula

Statins are commonly used in the middle-aged and elderly people for treatment of hyperlipidemia. Both alpha lipoic acid (ALA) and ellagic acid (EA) are natural antioxidants found in a normal diet. They can protect against cellular damage and induce cellular apoptosis in many types of cancer cells. Fluvastatin (FLV) was combined with ALA and EA in a nanostructured lipid carrier (NLC) formula. The prepared NLCs were imaged with a transmission electron microscope (TEM). Particle size and zeta potential and FLV entrapment efficiency (%EE) were measured, and the FLV release profile was constructed. Cellular viability, caspase-3 enzyme levels, and cellular cycle were analyzed. The prepared NLCs were spherical, with a size of 85.2?±?4.1 nm, and had a zeta potential of ??25.1?±?3.4 mV and a %EE of 98.2?±?1.1%. FLV IC₅₀ was decreased by half by the formula and by about 30% when compared with the three drugs together. According to cell-cycle analysis, treatment with FLV-ALA-EA NLCs caused a significant increase in pre-G1 phase by about 1.44-fold in comparison with FLV-ALA-EA. These findings demonstrate that ALA and EA induced cell death, which makes their combination with FLV a candidate for prostate cancer therapy.