Raw diets for dogs and cats: Potential health benefits and threats.

Raw meat-based diets for pet nutrition are becoming increasingly popular. The percentage of meat content, composition of nutrients, and amount of additives started to play an important role in the recipe of a given food. However, the use of healthier and unprocessed food must also be balanced with the animal's specific needs based on its anatomy, physiology, and behavior. There are many potential advantages and disadvantages of a biologically appropriate raw food (BARF) diet, and all of them should be considered before switching to this approach. Raw meat is considered a diet closest to nature and least processed. However, raw diets threaten pet health because of the potential for nutrient imbalances. The choice of raw meat in pets' everyday diet should be supported by the veterinarian's medical decision and preferably also with nutritionist help. Growing animals require a specific Ca:P ratio in their diet, which may be improper in raw meat. For cats, taurine levels must be carefully checked. In addition, an imbalanced raw-meat diet can be the cause of poor semen quality in males. Females are prone to inhibition of the estrus cycle, especially due to hyperthyroidism. Exogenous thyroid hormone intake is a real concern when feeding a neck/head meat with thyroid glands. There is also a possibility of bacterial or parasitic presence in raw meat. The present paper aims to summarize the current state of knowledge about the benefits and threats of eating a raw meat diet for the health concerns of companion animals.

Effect of prudent use of antimicrobials in the early phase of infection in pigs on the performance and meat quality of fattening pigs.

The presented study was a part of the ThermoEye project. The study examined the effect of prudent antibiotic treatment in response to illness on the fattening performance, slaughter traits, and meat quality of the pig fatteners. Pigs were divided into an experimental group - EXP and a control - CON. In both groups, the body temperature was monitored, and diseases were confirmed by a veterinarian. In the EXP group, metaphylaxis was used in each case of confirmed disease. The EXP fatteners had greater slaughter weight (by 4.7 kg) and meatiness (by 2.1 percentage points) compared to the CON pigs. The pH in pork was lower in EXP compared to CON pigs. The longissimus thoracis et lumborum of EXP pigs was characterised by a lower cooking loss (28.30 vs. 30.45%) and yellower colour compared to the CON group. Among the amino acids, only the content of histidine (by 5.2%; P < 0.01) and tyrosine (by 7.2%; P < 0.01) was significantly greater in the meat of the CON group compared to EXP, with no effect of sex noted. The content of SFA (by 14.6%; P < 0.05), UFA (by 15.6%; P < 0.05), MUFA (by 17.8%; P < 0.05), MCFA (by 14.1%; P < 0.05), and LCFA (by 15.6%; P < 0.05) was also greater in CON compared to EXP meat. In conclusion, automated monitoring of pig body temperature during the fattening period enables more precise, prudent treatment and effective animal health control while reducing costs due to disease losses and pharmacotherapy. It allows optimal production and affects meat quality.

Distinct dynamics of lipid accumulation by porcine cumulus cells during in vitro maturation with follicular fluid of low and high fatty acid contents.

Follicular fluid (FF) is a complex biological medium providing a fully balanced microenvironment for the oocyte. The standard medium for in vitro maturation of porcine oocytes contains 10% of FF which due to its unknown and inconstant composition is a source of a significant variation. This study aimed to investigate whether follicular fluids of significantly different fatty acid contents (standardized follicular fluid) supplemented to porcine IVM media affects lipid metabolism of porcine oocytes and cumulus cells. Two categories of FF from cyclic gilts containing high (H) and low (L) FA content was added to IVM medium for oocytes from prepubertal gilts. Altogether, 521 cumulus oocyte-complexes (oocytes and corresponding cumulus cells) were analyzed for mRNA expression of 7 genes regulating lipid metabolism and selected traits of the lipid droplets (LD). The applied FFs of different FA levels exerted distinct effects on oocytes and cumulus cells (CCs). During IVM oocytes tended to utilize the lipids as demonstrated by the reduced LD number and lipid fluorescence, whereas cumulus cells accumulated lipids as indicated by the increase in LD number, the occupied area and fluorescence level. Changes in cumulus cells were independent of the FA content in the follicular fluid which means an efficient lipid accumulation during IVM. Final analysis including the effect of FA level on LD traits in oocytes and corresponding CCs revealed two distinct patterns. COCs matured in FF of high FA content were characterized by elevated dynamics of lipid accumulation in CCs and stable lipid content in oocytes. In the case of FF with low FA content, CCs accumulated lipids at a significantly lower rate whereas lipid level in oocytes was reduced. The alterations observed in the LD parameters were not accompanied by changes in oocyte nuclear maturation and in transcript level of any from the 7 analyzed genes. In conclusion, fatty acid content of the follicular fluid supplemented to porcine IVM medium affects lipid metabolism in cumulus cells of the maturing oocyte and application of a standardized FF may help to improve the quality of porcine oocytes matured in vitro.

Review: Methanogens and methane production in the digestive systems of nonruminant farm animals.

The greenhouse gases (GHGs) derived from agriculture include carbon dioxide, nitrous oxide, and methane (CH4). Of these GHGs, CH4, in particular, constitutes a major component of the GHG emitted by the agricultural sector. Along with environmental concerns, CH4 emission also leads to losses in gross energy intake with economic implications. While ruminants are considered the main source of CH4 from agriculture, nonruminant animals also contribute substantially, and the CH4 emission intensity of nonruminants remains comparable to that of ruminants. Means of mitigating CH4 emissions from enteric fermentation have therefore been sought. Methane is produced by methanogens-archaeal microorganisms that inhabit the digestive tracts of animals and participate in fermentation processes. As the diversity of methanogen communities is thought to be responsible for the differences in CH4 production among nonruminant animals, it is necessary to investigate the archaeal composition of specific animal species. Methanogens play an important role in energy metabolism and adipose tissue deposition in animals. Higher abundances of methanogens, along with their higher diversity, have been reported to contribute to lean phenotype in pigs. In particular, a greater abundance of Methanosphaera spp. and early dominance of Methanobrevibacter smithii have been reported to correlate with lower body fat formation in pigs. Besides the contribution of methanogens to the metabolic phenotype of their hosts, CH4 release reduces the productivity that could be achieved through other hydrogen (H2) disposal pathways. Enhanced participation of acetogenesis in H2 disposal, leading to acetate formation, could be a more favorable direction for animal production and the environment. Better knowledge and understanding of the archaeal communities of the gastrointestinal tract (GIT), including their metabolism and interactions with other microorganisms, would thus allow the development of new strategies for inhibiting methanogens and shifting toward acetogenesis. There are a variety of approaches to inhibiting methanogens and mitigating methanogenesis in ruminants, which can find an application for nonruminants, such as nutritional changes through supplementation with biologically active compounds and management changes. We summarize the available reports and provide a comprehensive review of methanogens living in the GIT of various nonruminants, such as swine, horses, donkeys, rabbits, and poultry. This review will help in a better understanding of the populations and diversity of methanogens and the implications of their presence in nonruminant animals.

Evidence for a role of RUNX1 as recombinase cofactor for TCRß rearrangements and pathological deletions in ETV6-RUNX1 ALL.

T-cell receptor gene beta (TCRß) gene rearrangement represents a complex, tightly regulated molecular mechanism involving excision, deletion and recombination of DNA during T-cell development. RUNX1, a well-known transcription factor for T-cell differentiation, has recently been described to act in addition as a recombinase cofactor for TCRδ gene rearrangements. In this work we employed a RUNX1 knock-out mouse model and demonstrate by deep TCRß sequencing, immunostaining and chromatin immunoprecipitation that RUNX1 binds to the initiation site of TCRß rearrangement and its homozygous inactivation induces severe structural changes of the rearranged TCRß gene, whereas heterozygous inactivation has almost no impact. To compare the mouse model results to the situation in Acute Lymphoblastic Leukemia (ALL) we analyzed TCRß gene rearrangements in T-ALL samples harboring heterozygous Runx1 mutations. Comparable to the Runx1+/- mouse model, heterozygous Runx1 mutations in T-ALL patients displayed no detectable impact on TCRß rearrangements. Furthermore, we reanalyzed published sequence data from recurrent deletion borders of ALL patients carrying an ETV6-RUNX1 translocation. RUNX1 motifs were significantly overrepresented at the deletion ends arguing for a role of RUNX1 in the deletion mechanism. Collectively, our data imply a role of RUNX1 as recombinase cofactor for both physiological and aberrant deletions.

Technical note: Interchangeability and comparison of methane measurements in dairy cows with 2 noninvasive infrared systems.

This study aimed to compare measurements of methane (CH4) and carbon dioxide (CO2) concentrations in the breath of dairy cows kept in commercial conditions using the Fourier-transform infrared spectroscopy (FTIR) and nondispersive infrared spectroscopy (NDIR) methods. The measurement systems were installed in an automated milking system. Measurements were carried out for 5 d using both systems during milkings. The measurements were averaged per milking, giving 467 observations of CH4 and CO2 concentrations of 44 Holstein Friesian cows. The Pearson correlation between observations from the 2 systems was 0.86 for CH4, 0.84 for CO2, and 0.88 for their ratio. The repeatability of FTIR (0.53 for CH4, 0.57 for CO2, and 0.28 for their ratio) was somewhat higher than that of NDIR (0.57 for CH4, 0.47 for CO2, and 0.25 for their ratio). The coefficient of individual agreement was 0.98 for CH4, 0.89 for CO2, and 0.89 for their ratio; the concordance correlation coefficient was 0.48 for both gases and 0.24 for their ratio. We showed that FTIR and NDIR give similar results in commercial farm conditions. They can therefore be used interchangeably to generate a larger data set, which could then be further used for genetic evaluation.

TCRα rearrangements identify a subgroup of NKL-deregulated adult T-ALLs associated with favorable outcome.

T-cell acute lymphoblastic leukemia (T-ALL) results from leukemic transformation of T-cell precursors arrested at specific differentiation stages, including an 'early-cortical' thymic maturation arrest characterized by expression of cytoplasmic TCRß but no surface T-cell receptor (TCR) and frequent ectopic expression of the TLX1/3 NK-like homeotic proteins (NKL). We designed a TCRα VJC PCR to identify clonal TCRα rearrangements in 32% of 127 T-ALLs, including 0/52 immature/TCRγδ lineage cases and 41/75 (55%) TCRαß lineage cases. Amongst the latter, TCRα rearrangements were not identified in 30/54 (56%) of IMß/pre-αß early-cortical T-ALLs, of which the majority (21/30) expressed TLX1/3. We reasoned that the remaining T-ALLs might express other NKL proteins, so compared transcript levels of 46 NKL in T-ALL and normal thymic subpopulations. Ectopic overexpression of 10 NKL genes, of which six are unreported in T-ALL (NKX2-3, BARHL1, BARX2, EMX2, LBX2 and MSX2), was detectable in 17/104 (16%) T-ALLs. Virtually all NKL overexpressing T-ALLs were TCRα unrearranged and ectopic NKL transcript expression strongly repressed Eα activity, suggesting that ectopic NKL expression is the major determinant in early-cortical thymic T-ALL maturation arrest. This immunogenetic T-ALL subtype, defined by TCRß VDJ but no TCRα VJ rearrangement, is associated with a favorable outcome in GRAALL-treated adult T-ALLs.

Prepubertal heifers versus cows-The differences in the follicular environment.

The oocyte quality is to a large extent influenced by the sexual maturity of the donor female. Although this phenomenon has already been broadly described in domestic animals, the underlying mechanisms are poorly understood. Published data focus on oocyte ultrastructure, fertilization abnormalities, and blastocyst developmental rate. The goal of the present experiment was to characterize the follicular environment (oocyte, cumulus [CC] and granulosa (GC) cells as well as follicular fluid [FF]) in ovarian follicles of prepubertal heifers and cows. Each experimental replicate included the following set of traits within individual follicles: lipid droplets (LDs) number in oocytes, expression of seven genes involved in energy metabolism (fatty acids [FAs] metabolism-ELOVL2, ELOVL5, SCD, FADS2, glucose transport-GLUT1, GLUT3, GLUT8) in CC and GC as well as FA composition and glucose concentration in FF. According to our results, cow oocytes were larger in diameter and contained more LD than those from prepubertal heifers, both before and after IVM. The LD number was also higher in cow oocytes after IVM, when compared to immature oocytes. The FF from cow follicles had elevated glucose content similarly to the majority of the analyzed FA. Transcript analysis revealed differences for five out of seven analyzed genes (ELOVL, FADS2, SCD, GLUT3, GLUT8) in CC and GC cells. However after considering the female category, the only difference was noticed for the mRNA of SCD gene, which was more abundant in cow GC. This finding may indicate distinct roles of CC and GC in follicular energy metabolism. In conclusions, we suggest that distinct properties of follicular environment in prepubertal heifers and cows may be responsible for differences in the quality of oocytes from the two categories of donors. We hypothesize that suboptimal environment in heifer follicles (glucose and FA lower content in FF) determines reduced quality of their oocytes (lower diameter and LD number) and limited maturation potential. Besides, energy demands of heifer oocytes may be restricted due to a low LD number, exerting a negative effect on the development of the future embryo. The advantages of cow gametes (e.g., higher LD number and diameter) attributed to oocytes of superior quality may support the statement that cows donate oocytes of better quality than heifers.

Short communication: A nanoemulsified form of oil blends positively affects the fatty acid proportion in ruminal batch cultures.

Two consecutive rumen batch cultures were used to study the effect of nanoemulsified oils as a new type of supplement, on the in vitro fatty acid proportion and vaccenic acid formation. Three levels (3, 5, and 7%) of 2 different oil blends [soybean:fish oil (SF) or rapeseed-fish oil (RF)] were used. Both oil blends were used either in the raw form (SF or RF, respectively) or in the nanoemulsified form (NSF or NRF, respectively). The diets were the control (0%), which consisted of a dry total mixed ration without any supplements, the control plus 3, 5, or 7% of the SF or RF oil blend in appropriate form (raw or nanoemulsified). For each treatment, 6 incubation vessels were used. Each batch culture was incubated for 24h and conducted twice in 2 consecutive days. All supplements were calculated as a percentage of the substrate dry matter (400mg). Nanoemulsified supplements were recalculated to make sure the oil amount was equal to the raw oil supplementation levels. The results from both experiments indicated that the proportions of vaccenic acid and cis-9,trans-11 C18:2 increased when a raw oil blend was supplemented; on the other hand, no influence of nanoemulsified form of oil blend was observed on the proportion cis-9,trans-11 C18:2. Generally, supplementation with the nanoemulsified oil blends had less effect on biohydrogenation intermediates than the raw form of oil blends. However, the nanoemulsified form had a greater effect on the increase of n-3 and n-6 fatty acids. Nanoemulsified oil blends had a positive effect on decreasing the transformation rate of polyunsaturated fatty acids to saturated fatty acids in the biohydrogenation environment. Supplements of nanoemulsified oil blends tended to be more effective than supplements of raw oils in preserving a greater proportion of polyunsaturated fatty acids in the fermentation culture.

Protein expression parallels thermal tolerance and ecologic changes in the diversification of a diving beetle species complex.

Physiological changes associated with evolutionary and ecological processes such as diversification, range expansion or speciation are still incompletely understood, especially for non-model species. Here we study differences in protein expression in response to temperature in a western Mediterranean diving beetle species complex, using two-dimensional differential gel electrophoresis with one Moroccan and one Iberian population each of Agabus ramblae and Agabus brunneus. We identified proteins with significant expression differences after thermal treatments comparing them with a reference EST library generated from one of the species of the complex (A. ramblae). The colonisation during the Middle Pleistocene of the Iberian peninsula by A. ramblae, where maximum temperatures and seasonality are lower than in the ancestral north African range, was associated with changes in the response to 27 °C in proteins related to energy metabolism. The subsequent speciation of A. brunneus from within populations of Iberian A. ramblae was associated with changes in the expression of several stress-related proteins (mostly chaperons) when exposed to 4 °C. These changes are in agreement with the known tolerance to lower temperatures of A. brunneus, which occupies a larger geographical area with a wider range of climatic conditions. In both cases, protein expression changes paralleled the evolution of thermal tolerance and the climatic conditions experienced by the species. However, although the colonisation of the Iberian peninsula did not result in morphological change, the speciation process of A. brunneus within Iberia involved genetic isolation and substantial differences in male genitalia and body size and shape.

The metabolic profile of growing lambs fed diets rich in unsaturated fatty acids.

The effect of two diets enriched with unsaturated fatty acids--one containing the addition of dried distillers grains with solubles (DGS) and the other the addition of false flax--Camelina sativa cake (CS)--on some metabolic parameters and hormone concentration in growing lambs was determined in this experiment. A total of 21 ram lambs of the Polish Whiteheaded mutton sheep were divided into three groups (the control, receiving DGS and CS). The diets were administered to animals for 6 weeks. During the experiment, peripheral blood was collected. Glucose (GL), total cholesterol (CH), triglycerides (TG), free fatty acids (FFA), insulin (IN), leptin (LE), triiodothyronine (T3) and thyroxine (T4) were assayed in serum. The age-dependent reduction in CH and TG limited by both experimental diets were observed. A significant increase in FFA concentration was observed in samples collected in the last, that is, third, time period. This was most probably caused by a 12-h pre-slaughter fasting period. A significantly lower dynamic of FFA increase in that experimental period was found in animals receiving the experimental feed. Insulin concentration in DGS-receiving lambs was increased, in contrast to the CS-receiving lambs, in which it was lower when compared to the control. LE concentration was decreased by both experimental diets, more markedly in the DGS-receiving animals. No differences between the experimental groups and the control were observed in T3 and T4 concentrations. The effect of 12-h pre-slaughter fasting was statistically highly significant for the levels of examined blood markers and hormones, except for TG and IN in the group of lambs receiving the experimental diet with CS.

Plant components with specific activities against rumen methanogens.

A wide range of plant bioactive components (phytochemicals) have been identified as having potential to modulate the processes of fermentation in the rumen. The use of plants or plant extracts as natural feed additives has become a subject of interest not only among nutritionists but also other scientists. Although a large number of phytochemicals (e.g. saponins, tannins and essential oils) have recently been investigated for their methane reduction potential, there have not yet been major breakthroughs that could be applied in practice. A key tenet of this paper is the need for studies on the influence of plant components on methane production to be performed with standardized samples. Where there are consistent effects, the literature suggests that saponins mitigate methanogenesis mainly by reducing the number of protozoa, condensed tannins both by reducing the number of protozoa and by a direct toxic effect on methanogens, whereas essential oils act mostly by a direct toxic effect on methanogens. However, because the rumen is a complex ecosystem, analysis of the influence of plant components on the populations of methanogens should take into account not only the total population of methanogens but also individual orders or species. Although a number of plants and plant extracts have shown potential in studies in vitro, these effects must be confirmed in vivo.

No single way to explain cytoplasmic maturation of oocytes from prepubertal and cyclic gilts.

The objective of this study was to evaluate selected aspects of cytoplasmic maturation in oocytes from prepubertal and cyclic crossbred gilts before and after in vitro maturation. For this purpose, cortical granule redistribution, mitochondrial DNA content and mitochondria translocation were analyzed. Moreover, for the first time the fatty acid profiles in follicular fluid (FF) of both gilt categories was evaluated. The nuclear maturation (the percentage of metaphase II oocytes was 83% in prepubertal gilts compared with 87% in cyclic gilts), cortical granule relocation from the cortex to peripheral ooplasm (98.7% vs. 98.8% of oocytes, respectively) and mitochondrial DNA content (227 543 vs. 206 660, respectively) was not affected by sexual maturity of the donor gilt. However, the redistribution of active mitochondria during in vitro maturation was observed only in the oocytes of cyclic gilts. With regard to FF analysis, saturated, unsaturated, and monounsaturated fatty acids were significantly more abundant in the FF of prepubertal females. In particular, stearic (C18:0) and palmitic (C16:0) fatty acids had significantly higher concentrations in the FF of prepubertal gilts. In conclusion, although the oocytes of prepubertal gilts matured in vitro at a rate similar to those of cyclic gilts, they differed with respect to the selected factors attributed to cytoplasmic maturation. We suggest that the higher content of particular fatty acids, which is known to have a negative influence on oocyte maturation, as well as impaired mitochondria redistribution are factors limiting the maturation potential of oocytes from prepubertal gilts.

Application of microbial assay for risk assessment biotest in evaluation of toxicity of human and veterinary antibiotics.

The microbial assay for risk assessment (MARA) is a multispecies, growth inhibition microplate toxicity test with 11 microorganisms individually lyophilized in microplate wells. The microbial species representing wide diversity, generated 11 microbial toxic concentration (MTC) values were obtained giving a unique "toxic fingerprint" profile of the test sample. The toxicity of 14 antibiotics was evaluated with the MARA test. The fingerprints for each group of antibiotics differ, indicating a particular toxic profile. Tetracyclines were the most toxic antibacterials with the minimum MTC value of 3.6 µg L(-1). In the group of tetracyclines the order of the three most sensitive microbial strains was the same 2 > 6 > 7. Quinolones affected the most sensitive strain(s) at concentrations of 12-75 µg L(-1). The MARA bioassay exhibited different spectra of toxic responses to microbial strains for the first and second generation quinolones. However, for first generation quinolones strain 6 was substantially more sensitive than the other microorganisms, while second generation quinolones were most toxic to strain 3, followed by 6 then 4. In this instance, the three strains belong to two different phylogenetic groups-strain 3 is γ-proteotype and strains 4 and 6 are ß-proteotype.

Repeated low-dose ultraviolet (UV) B exposures of humans induce limited photoprotection against the immune effects of erythemal UVB radiation.

BACKGROUND: Exposure of human subjects to ultraviolet (UV) B radiation causes immunosuppression. Most experiments to date have not tested the effects of low daily doses of UVB radiation. OBJECTIVES: To ascertain whether photoprotection against several UV-induced immune effects might develop following repeated exposure. METHODS: Groups of approximately 30 healthy individuals were given whole-body UVB irradiation on each of 10 consecutive days with 0.7 minimal erythema dose, or whole-body irradiation as before followed by a single erythemal UVB dose on a small body area, or irradiated only with a single erythemal UVB dose on a small body area, or were not irradiated. They were sensitized with diphenylcyclopropenone (DPCP) 24 h after the final dose, and skin biopsies collected to assess cytokine mRNA expression and the number of cells with thymine dimers and expression cyclooxygenase (COX)-1 and COX-2. RESULTS: The contact hypersensitivity (CHS) response to DPCP was significantly lower in the three irradiated groups compared with the unirradiated controls, while cutaneous interleukin (IL)-1beta, IL-6, IL-10 and tumour necrosis factor-alpha mRNAs, COX-1 and COX-2 and thymine dimers were all significantly higher. When the single erythemal UVB dose was given following the repeated low exposures, a slight downregulation in cytokine expression and thymine dimer formation was indicated. CONCLUSIONS: The repeated low doses of UVB protected to a limited extent against the effects of an erythemal UVB dose on cytokine expression and thymine dimer formation, but not on CHS or COX enzymes.

Similar PAI-1 expression in visceral and subcutaneous fat of postmenopausal women.

Recent studies showed that intraabdominal visceral fat located in the mesenterium and omentum may significantly influence the circulating plasminogen activator inhibitor type 1 (PAI-1). To substantiate this link, we performed analysis of PAI-1 expression in human visceral and subcutaneous adipose tissues in peri- and postmenopausal women. The samples of both visceral and subcutaneous fat from 28 generally healthy women (aged 45-69 years) with a wide range of body mass index (BMI; 22.30-38.67 kg/m2), who underwent surgical operation due to benign ovary and uterine tumours, were obtained. In these samples, expression of mRNAs for PAI-1, tumour necrosis factor alpha (TNFalpha), acyl-CoA synthetase (ACS), and glucose transporter (GLUT-4) was analysed by relative quantitative RT PCR and correlated with plasma PAI-1 antigen. In addition, visceral fat area was measured with computer tomography. Both types of fat tissues contained similar quantities of PAI-1 mRNA, and there was no correlation between plasma PAI-1, measured both by antigen and activity, with either visceral or subcutaneous fat PAI-1 mRNA. Furthermore, there was no significant association between the expression of PAI-1 mRNA and TNFalpha mRNA in tested fat samples. PAI-1 mRNA in both fat tissues was significantly correlated with plasma levels of estradiol (positive correlation) and follicle-stimulating hormone (FSH; negative correlation). Finally, the expression of PAI-1 mRNA was negatively correlated with mRNA of ACS present in both fat tissues. In summary, this study directly indicates that PAI-1 mRNA is similarly expressed in both subcutaneous and visceral fat of peri- and postmenopausal women and its expression strongly depends upon lipid metabolism.

The moderating role of hardiness and social support in the relation between job stressors and well-being. A lesson from a clerical women sample.

The study concerns the moderating role of two variables--hardiness and social support--in the relation between job stressors and well-being. It was checked if there is a connection between hardiness and social support, and if these variables are directly related to the level of well-being. It was also hypothesized that hardiness and social support buffer the negative influence of stressors on well-being. The fact that social support and well-being are multidimensional variables was taken into account. The cross-sectional study was conducted on a group of 200 women employed as office workers. The results showed that hardiness correlates with the indexes of emotional support as well as practical support coming from supervisors. The connection, however, was not strong. Hardiness and social support were directly connected with the level of well-being. Interaction of these two independent variables played a marginal role. As far as the buffering role of these variables was concerned, the results have shown that hardiness did not play a buffering role and social support did to a limited degree only. The results of the study proved that it is justifiable to treat social support and well-being as multidimensional variables.

Dual regulatory effects of nitric oxide on plasminogen activator inhibitor type 1 expression in endothelial cells.

In this report we compared the mechanism by which nitric oxide (NO), generated exogenously and endogenously, affects the plasminogen activator inhibitor type 1 (PAI-1) expression in endothelial cells. For this purpose, we stimulated the endothelial cell line EA.hy 926 with tumour necrosis factor alpha (TNFalpha) in the presence of the exogenously NO-releasing donors, sodium nitroprusside (SNP) and S-nitroso-N-acetylpenicillamine, or regulators of nitric oxide synthase (NOS) inhibitor N-nitro-L-arginine-methyl ester hydrochloride and substrate L-Arg. Expression of PAI-1 in EA.hy 926 cells was determined by measuring the level of mRNA, using relative quantitative reverse transcriptase PCR, and protein, using ELISA. In addition, we estimated the level of activation of two mitogen-activated protein kinases (MAPKs), extracellular signal-regulated kinase (ERK1/2) and c-Jun N-terminal kinase (JNK1/2), in the cells before and after treatment with TNFalpha, in the presence or absence of NO donors and inhibitors. In contrast to exogenously released NO that significantly reduced mostly basal PAI-1 expression, endogenously generated NO by NOS potentiated TNFalpha-induced upregulation of PAI-1 expression. Exogenously and endogenously generated NO causes different effects on activation of the MAPKs ERK1/2 and JNK1/2. Specifically, the SNP-released NO activates only ERK1/2, while endogenously generated NO in a pathway induced by TNFalpha activates both MAPKs. Thus our data indicate that due to different cellular locations and mechanisms of generation, NO may participate in various signalling pathways leading to opposite effects on PAI-1 expression in endothelial cells.