Reduced-Oxidized Glutathione Status as a Potential Index of Oxidative Stress in Mature Cereal Grain

The purpose of this study was to examine the reduced and oxidized glutathione status of selected cereal grains as a potential index of balance between oxidative stress and antioxidant systems, and the contribution of reduced glutathione to the total antioxidant status in cereal grain extracts. Wheat cultivars Almari and Henika, barley cultivars Gregor and Mobek, rye cultivar Dañkowskie Złote, oat cultivar Sławko, and buckwheat cultivar Kora were used. Total antioxidant status (TAS) was measured by the ABTS (2,2'-azinobis(3-ethyl-benzothiazoline-6-sulphonate)) method. Contents of total phenolic compounds were also determined. Reduced (GSH) and oxidized glutathione (GSSG) (γ-glutamyl-cysteinyl-glycine) were assayed using the spectrofluorimetric method, and results were confirmed by the enzyme recycling method. Correlation coefficient for the GSH/GSSG ratio was r = 0.79. Correlation between TAS and the total phenolic compound content was r = 0.81. Correlation between GSH/GSSG ratio and TAS values was r= 0.46, depending on the extraction system used. The GSH/GSSG ratio may indicate a hierarchy among different cultivars and variance of cereal grains against damage caused by reactive oxygen species. For the main water-soluble antioxidants, our data indicate a potential hierarchy of resistance in investigated cereals against oxidative stress (buckwheat > wheat > barley ≈ rye > oat). This hierarchy was confirmed by the ability of investigated cereal extracts to scavenge superoxide anion radicals in vitro. The reduced-oxidized glutathione status in different cereal grains can be applied as a potential index of balance between oxidative stress and antioxidant systems.     

NIR spectroscopy and partial least-squares regression for determination of natural alpha-tocopherol in vegetable oils

Near-infrared (NIR) spectroscopy and partial least-square regression were used for determination of alpha-tocopherol in edible oils after extraction with ethanol. The standard error of calibration and the standard error of prediction were calculated for evaluation of the calibration models. The chemometric calibration model was prepared in spectral region 6500-4500 cm(-1) for standard alpha-tocopherol solutions (0.54-53.54 mg/mL). Obtained mean concentrations of natural alpha-tocopherol in different types of oils varied from 17.53 to 57.10 mg/100 g. Net analyte signal calculation was used to estimate detection limit (DL = 0.12 mg/mL), quantification limit (QL = 0.40 mg/mL), sensitivity (SEN = 0.045 mg/mL), and selectivity (SEL ranged between 0.24 and 0.54% of the measured reflectance signal) of the proposed NIR method. The comparable precision (RSD = 0.68-2.80% and 0.79-3.06%) and accuracy (recovery, 97.2-102.4% and 96.8-103.2%) for the proposed NIR and standard HPLC methods, demonstrate the benefit of the NIR method in the routine analysis of alpha-tocopherol in vegetable oils.     

Introduction of the entomopathogenic fungusEntomophaga maimaiga hum.,

The entomopathogenic fungusEntomophaga maimaiga was introduced into aL. dispar population situated in South Bulgaria for the first time in Europe. It was found that of all larvae collected in the five experimental plots, 6.3 % contained conidia and resting spores ofE. maimaiga, 14.2 % contained parasitoids and 2.3 % were infected with nucleopolyhedrovirus (NPV). The presence of the fungus in cadavers collected from each experimental plots and on five of the six collection dates was observed.     

Evaluation of the ability of colistin,amoxicillin (components of Potencil®), and fluoroquinolones to attenuate bacterial endotoxin‐ and Shiga exotoxin‐mediated cytotoxicity—In vitro studies

Escherichia coli is one of the major pathogens in humans and animals causing localized and systemic infections, which often lead to acute inflammation, watery diarrhea, and hemorrhagic colitis. Bacterial lipopolysaccharide (LPS) and Shiga exotoxins (Stx) are mostly responsible for such clinical signs. Therefore, highly effective treatment of E. coli infections should include both eradication of bacteria and neutralization of their toxins. Here, for the first time, we compared the in vitro ability of common antibiotics to decrease LPS‐ and Stx‐mediated cytotoxicity: colistin, amoxicillin (used separately or combined), enrofloxacin, and its metabolite ciprofloxacin. Three experimental scenarios were realized as follows: (a) the direct effect of antibiotics on endotoxin, (b) the effect of antibiotic treatment on LPS‐mediated cytotoxicity in an experiment mimicking “natural infection,” (c) the effect of antibiotics to decrease Stx2e‐mediated cytotoxicity. Two cell lines, A549 and Vero cells, were used to perform cytotoxic assays with the methyl tetrazolium (MTT) and lactate dehydrogenase leakage (LDH) methods, respectively. Colistin and amoxicillin, especially used in combination, were able to attenuate LPS toxic effect, which was reflected by increase in A549 cell viability. In comparison with other antibiotics, the combination of colistin and amoxicillin exhibited the highest boster or additive effect in protecting cells against LPS‐ and Stx2e‐induced toxicity. In summary, in comparison with fluoroquinolones, the combination of colistin and amoxicillin at concentrations similar to those achieved in plasma of treated animals exhibited the highest ability to attenuate LPS‐ and Stx2e‐mediated cytotoxicity.     

Survivability and developmental competences of domestic cat (Felis catus) oocytes after Cryotech method vitrification

The aim of this study was to evaluate the applicability of the Cryotech technique for the vitrification of domestic cat (Felis catus) oocytes, as a model for other feline species threatened with extinction. This technique, in which oocytes are stored in a minimal volume of medium, is already widely used in human assisted reproductive technology. In the first part of this study, a viability test (EtBr/FDA) was used to evaluate the toxicity of the vitrification media (solutions). After IVM, oocytes were placed in vitrification and warming solutions according to the manufacturer's procedure, with or without exposure to liquid nitrogen. The solutions and the vitrification procedure each caused a reduction in oocyte viability, with survival rates of 71.4% in oocytes exposed to the Cryotech media (without cooling in liquid nitrogen), and 62% in oocytes that were vitrified. In the second part of the experiment, parthenogenetic activation was used to evaluate the developmental potential of oocytes previously vitrified using the Cryotech method. After warming, the oocytes were activated using a combination of 0.7 µM ionomycin in TCM 199 medium (5 min) followed by 2 mM 6-DMAP in TCM 199 supplemented with 10% FBS (3 hr), then cultured and evaluated every 24 hr for parthenogenetic cleavage. In the experimental group, 23/50 (46%) cleaved embryos were obtained. Domestic cat oocytes, vitrified by the Cryotech method, are characterized by high survival rates. However, it is necessary to improve the technique to increase the developmental competence of embryos obtained from vitrified oocytes.     

Maternal HMB treatment affects bone and hyaline cartilage development in their weaned piglets via the leptin/osteoprotegerin system

It has been demonstrated in animal studies that prenatal administration of β‐hydroxy‐β‐methylbutyrate (HMB, metabolite of leucine) influences general growth and mechanical endurance of long bones in newborn offspring in sex‐dependent manner. The present experiment was conducted to evaluate the effect of HMB treatment of pregnant sows on bone development in offspring at weaning. From 70th day until the 90th day of gestation, sows received either a basal diet (n = 12) or the same diet supplemented with HMB (n = 12) at the dose of 0.2 g/kg of body weight/day. Femora obtained from six males and females in each group weaned at the age of 35 days were examined. Maternal HMB treatment significantly enhanced body weight and changed bone morphology increasing femur mechanical strength in both sexes. Maternal HMB supplementation also elevated bone micro‐ and macroelement concentrations and enhanced content of proteoglycans in articular cartilage. Based on the obtained results, it can be concluded that maternal HMB supplementation in the mid‐gestation period significantly accelerated bone development in both sexes by upregulation of a multifactorial system including leptin and osteoprotegerin. However, the sex (irrespective of the HMB treatment) was the factor which influenced the collagen structure in cartilages and trabecular bone, as demonstrated both by the Picrosirius red staining and performed analysis of thermal stability of collagenous tissues. The structural differences in collagen between males and females were presumably related to a different collagen maturity. No studies conducted so far provided a detailed morphological analysis of bone, articular cartilage, growth plate and the activities of the somatotropic and pituitary–gonadal axes, as well as leptin/osteoprotegerin system in weaned offspring prenatally treated with HMB. This study showed also the relationship between the maternal HMB treatment and bone osteometric and mechanical traits, hormones, and growth and bone turnover markers such as leptin, osteoprotegerin and insulin‐like growth factor‐1.     

Analysis of the factors creating consumer attributes of roasted beef steaks

The aim of the study was to analyze the factors creating consumer attributes of roasted beef steaks of various animals. Eight cuts from 30 carcasses (characterized by various types of animal, conformation and fat class, rib fat thickness, ossification score) were selected. Samples were prepared using the roasting method and consumers rated the tenderness, juiciness, flavor, overall acceptability (rated in a 100‐point scale), and satisfaction (rated from 2 to 5) for analyzed samples. No influence of type of animal, fat class, conformation class or ossification score on the results of consumer analysis was observed. For all analyzed factors, the influence of cut on consumer analysis was observed (the highest values of all consumer attributes were observed for tenderloin ‐ for juiciness significantly higher than for other cuts, for tenderness, flavor and MQ4 comparable only with rump (RMP231), while for overall acceptability and satisfaction – with both rump cuts). For rib fat thickness consumer attributes of roasted beef meat were not linear, but the influence was observed – the highest values of consumer attributes were observed for 13 mm rib fat thickness.     

Universal rules for fragmentation of land by humans

The morphology of parcel patterns created by humans both in urban and rural areas is investigated. The parcel size distribution function, f(a), provides a criterion, that enables unambiguous classification of each piece of land as city core, suburbs, or rural area. The morphology of the rural area corresponds to a scale-free structure and follows a power-law distribution f(a) ~ a ⁻ⁿ of the parcel areas with the exponent n ≈ 1. In suburbs, the area distribution follows the log-normal distribution. In the city core, f(a) has an unimodal shape with an algebraically decaying tail, n = 2. Our study is based on data originating mainly from North America, the Hawaiian Islands, and Australia. For the regions analyzed, the characteristics of the parcel size distribution are universal and robust with respect to geographical, historical, and economical conditions accompanying development of a given area. The urbanization process can be described in terms of the changes of the morphology of the patterns of land fragmentation. In this formulation, the rural morphology, which can be thought as natural one because it exhibits a scale-free distribution of parcel sizes, is transformed into the artificial morphology developed in the city centers. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Intracardiac electrophysiological conduction parameters in adult dogs

Background: Invasive electrophysiology study (EPS) is one of the most important adjunct tests for the evaluation of arrhythmias in human medicine. However, EPS is neither well known nor widely used in veterinary medicine.

Objective: To define the values for intracardiac conduction parameters determined during invasive EPS in dogs.

Animals and methods: The study included 16 admitted dogs of various breeds, sex and ages and 6 control Beagles. In the Beagles, EPS was performed twice at 6-month intervals in order to verify the reproducibility of the results.

Results: No significant differences were found between the results of the baseline and repeated EPS performed in the Beagles. We found retrograde conduction in 13 (59%) out of 22 dogs and including 4 (31%) animals with concomitant ‘jumps’ in the atrioventricular node conduction curve, pointing to the presence of dual conduction in this node. The mean values of the AV and retrograde VA Wenckebach points equaled to 220 and 360 ms, respectively, suggesting that the capability of the descending route is higher than that of the ascending route.

Conclusion: The values determined in this study may be helpful in early detection of abnormalities in the electrical conduction system of the heart.