Volume 7, Issue 6

Caspases are proteases most notably involved in apoptosis and inflammation. Although mRNA content is better described, the constitutive protein content of procaspases between tissue types is not well documented. Since mRNA and protein content do not necessarily correlate, we aimed to discern protein content differences between various tissues. Protein content of procaspase-1, -8, -9, and -12 was assessed in gastrocnemius, heart, liver, and kidney. Since highly expressed in skeletal muscle, content of procaspase-12 was also analyzed in muscles with different fiber type compositions to discern any fiber type differences. Furthermore, Western analysis for procaspase-12 revealed prominent bands of ∼40 kDa and ∼30 kDa under basal conditions, in addition to the 50 kDa band corresponding to the full-length procaspase. Therefore, the content of these caspase-12 related species in the tissue and muscle types is also described. Results show protein content of procaspase-1,-8, -9, and -12 and caspase-12 related species differs between tissue types and do not necessarily correlate with mRNA content reported in the published literature. Procaspase-12 content in skeletal muscle may be fiber-type dependent with higher expression in more oxidative fibers. Furthermore, the 40 kDa species of caspase-12 was the dominant form of the protein in most tissues analyzed.

The aim of this study was to examine the effect of activators of protein kinase A — 3-isobutyl-1-methyl-xanthine (IBMX) and dibutyryl cyclic adenosine monophosphate (dbcAMP) — and of oxytocin (OT) on rabbit female reproductive function. We used equine chorionic gonadotropin (eCG) to improve follicular development and rabbit estrous synchronization and human chorionic gonadotropin (hCG) to induce ovulation. In the experimental group, the females were stimulated using gonadotropins together with either IBMX, dbcAMP or OT. In the animals kept until parturition, the conception rate, parturition rate, and numbers of stillborn and weaned pups were recorded. In the animals euthanized 18–19 hours after insemination, the eggs were flushed from the oviducts and cultured up to the blastocyst cell stage. Numbers of corpora lutea, zygotes, morulas and blastocysts were determined. Both dbcAMP and OT, but not IBMX, decreased conception and parturition rate. Both IBMX and OT, but not dbcAMP, decreased pup mortality rate. All three tested substances increased the weaning rate. Both IBMX and dbcAMP, but not OT, increased the numbers of corpora lutea, zygotes, and embryos at morula and hatching blastocyst stages. These observations confirm the stimulatory role of the protein kinase A-dependent signaling pathway activated by IBMX and dbcAMP in rabbit reproduction. OT may decrease pups mortality.

Male and female rats were fed diets supplemented with cholesterol and palm fat at 10 and 50 g/kg, respectively; serum, hepatic tissue and faeces were analysed. Cholesterol supplementation significantly increased serum and hepatic cholesterol both in male and female rats. Male and female rats fed the cholesterol-containing diet differed significantly in serum cholesterol concentration (2.48 µmol/mL vs 2.92 µmol/mL), concentration of serum triacylglycerols, but not in hepatic cholesterol concentration. The serum and hepatic cholesterol concentrations correlated non-significantly in male rats (r=0.491; P=0.063) and significantly in female rats (r=0.818; P<0.001). Cholesterol supplementation non-significantly decreased relative expression of the hepatic LDL receptor gene and significantly increased relative expression of the hepatic cholesterol 7α-hydroxylase gene in rats of both genders. The faeces of control rats contained similar amounts of cholesterol and bile acids. Cholesterol supplementation increased cholesterol concentration 10 times in the faeces of male rats and 12 times in faeces of female rats. The corresponding increases of bile acid concentration were much lower (83% in male rats and 108% in female rats). It can be concluded that the effects of cholesterol supplementation were more pronounced in female than in male rats.

Kynurenic acid (KYNA), a metabolite of the essential amino acid L-tryptophan, is a broad spectrum antagonist of excitatory amino acid receptors, which have also anticonvulsant and neuroprotective properties. After spinal cord injury (SCI), excitotoxicity is considered to play a significant role in the processes of secondary tissue destruction in both grey and white matter of the spinal cord. In this study, we have tested the potential therapeutic effect of glucosamine-kynurenic acid, administered after experimental compression-induced SCI in the rat. Spinal application of glucosamine-kynurenic acid continually for 24 hr after experimental SCI resulted in improved motor function recovery, beginning from the first week of evaluation and continuing until the end of the study (4 weeks). After 4 weeks’ survival, quantitative morphometric analysis of the spinal cord showed that glucosamine-kynurenic acid treatment was associated with improved tissue preservation at the lesion site. These findings indicate that spinal application of glucosaminekynurenic acid is neuroprotective and improves the outcome even when administered after spinal trauma. Our results suggest that the treatments initiated in early posttraumatic period can alleviate secondary injury and improve the final outcome after SCI.

Testosterone level has an influence on cognitive functions, especially spatial abilities. The relationship is, however, bidirectional and brain activity also affects testosterone levels. The aim of this study was to analyze the effects of an intensive 3D mental rotation task on testosterone levels in young healthy men and women. In the mental rotation task, men reached a higher top score (P=0.027) and total score (P=0.004) compared to women. In 8 out of 9 women (P=0.008) but not in men (P=0.129) testosterone levels decreased after one hour of mental rotation testing. A significant gender difference was shown (P<0.0001). In all women, plasma cortisol levels was significantly lower after testing (P=0.004). In men cortisol levels decreased in 7 out of 9 subjects (P=0.039). A significant gender difference was not found (P=0.19). No association was found in women between baseline testosterone levels and mental rotation total score (P=0.810). In men there was a positive correlation between baseline testosterone and mental rotation total score (P=0.015). A significant difference gender difference was seen in the association between testosterone and mental rotation score (P<0.05). Mental rotation stimuli caused significant changes in hormonal levels of testosterone and cortisol. A gender-specific response was detected in testosterone fluctuation.

Tacrine is a medication applied in cases of mild to moderate dementia in Alzheimer’s disease. By blocking acetylcholinesterase activity the drug increases the concentration of acetylcholine, whose effects influence the functions of different organs and systems of the body. The effect of tacrine on smooth muscle preparations isolated from rat stomach was studied by isometric registration of muscle contractility. Our investigations found a specific significant systematic decrease in the strength of consecutive tacrine-induced contractions of smooth muscle preparations, a phenomenon known as tachyphylaxis. The tacrineinduced tachyphylaxis was significantly inhibited by SQ22536 (inhibitor of adenylate cyclase activity), by blockers of nitric oxide synthase and KT5823 (inhibitor of protein kinase G). The process was not influenced by cyclopiazonic acid (specific blocker of sarco/endoplasmic reticulum Ca2+-ATPase,) and atropine (blocker of M-cholinergic receptors). We hypothesize that the overlapping and different time-development of the two opposing processes: smooth muscle contraction caused by acetylcholinesterase inhibition and tacrine-induced relaxation influenced by synthesis of nitric oxide, results in tachyphylaxis.

As canopy structure produces spatial heterogeneity of litter microclimatic conditions and thus is a crucial factor affecting ground insects, we hypothesized that low canopy openness has a positive effect on the activity of ground insect predators in forest and non-forest habitats. Blowfly larvae were used as bait along the canopy openness gradient (forest interior, forest edge, base of a solitary tree and meadow) and the attack rate was assessed after 30 min of exposure. Although the predation rate has a varying pattern throughout the year in different habitats, in contrast to previous studies, we observed a significant positive trend in predation rates toward the forest interior. A significant trend in predation rate was not observed in non-forest areas. We found that the trend was strongly influenced by ants as the most active taxon of predators (65%) attacking our baits, whereas ground beetles, the second-most active predators (21%), showed the opposite trend along the canopy openness gradient.

The effect of 19 environmental variables on species richness of macrophytes was studied in 39 Slovak streams. The studied streams were poor in species; in total, 88 macrophyte taxa were found and the average number of macrophytes per sampling site was 4, ranging from 0 to15. The most frequently occurring macrophytes were filamentous algae (occurrence at 38.6% of sampling sites), followed by Rhynchostegium riparioides (28.4%) and Phalaris arundinacea (19.3%). The strongest environmental gradient in the sampling site detected by factor analysis (factor 1 explains more than 32% variability) is related to the portion of artificial banks, shading by woody vegetation along banks, flexuosity of stream course and the portion of natural land cover in the contact zone of the stream, and can be interpreted as a natural-anthropogenic gradient. The following variables had the highest correlations with species richness of macrophytes: shading by woody vegetation (r=−0.507), portions of artificial bank (r=0.488), flexuosity (r=−0.457) and distance from stream source (r=0.388).

The influence of different chelates applied in the soil primary on Al and secondary on Fe and Mn mobilization and their removal from solution was investigated. The work compared the efficiency of 10 mM tartaric acid and 3 mM EDTA in soil washing process and accumulation potential of Pistia stratiotes in rhizofiltration process. The plant response on the toxic element Al and other elements Fe and Mn was determined through the nitrogen and free amino acids content in plants. The efficiency of chelates decreased in order 10 mM tartaric acid > deionized water > 3 mM EDTA for all studied elements. P. stratiotes was able to remove up to 90% of elements during the 15 days period. Higher content of toxic element Al and potential toxic elements Fe and Mn were observed in the roots than in the leaves with the increased time. The trend of Al accumulation correlated with Fe accumulation (R2=0.89). Toxicity impact of high level of Al was observed by increased free amino acids (AA) level. Proline, histidine, glutamic acid and glycine were the most synthesised free AA in leaves. Total AA content in leaves was significantly higher under chelates addition compared to control.

Toxic effects of acidic root medium and aluminium were evaluated in two forage cultivars of Lotus corniculatus differing in their tolerance to Al stress. The structural response of most of the root cells exposed to low pH without Al3+ differed markedly from that induced by the combined stress. Conspicuous alteration of the nucleus was present only at low pH 4.0 and disintegration of the cytoplasmic components was more drastic than in the roots exposed to acidic solution containing Al3+. Cells exposed to low pH without Al, did not produce wall thickenings. Severely damaged cytoplasm and localized death in some cortical cells or groups of cells contrasting with almost intact cells exposed to Al3+ stress were found. In this respect, a strong correlation between the occurrence of cell wall thickenings and a better preserved structure of the cytoplasm was observed. The frequency of cell damage in the more tolerant cultivar UFRGS was generally lower, significantly more cortical cells capable of maintaining their resting membrane potential were present than in the sensitive INIA Draco. The difference in their tolerance is related rather to the exudation of citrate and oxalate that was higher in UFRGS than to the accumulation of tannins, which increased after Al treatment in both cultivars.

This study investigates how yellow bluestem affects biodiversity in a typical Pannonian grassland. Beta diversity (i.e. the finescale spatial variability of species compositions), was estimated by the realized number of species combinations sampled at various scales. Sampling was performed by a standard protocol. Presences of plant species were recorded along 52.2 m long belt transect of 1044 units of 0.05x0.05 m contiguous microquadrats. According to the results the massive presence of tested C4 grass significantly reduced species richness of the grassland. Beta diversity assessment revealed that 90% of species combinations were lost due to yellow bluestem invasion. Fine-scale spatial pattern analyses showed complete local extinctions of other species from microsites dominated by yellow bluestem. This local extinction is enhanced by the specific clonal architecture of this species and by the accumulation of litter. Other dominant grasses had no effect on fine scale diversity, i.e. they could coexist well with other elements of the local flora. This study presents currently developed microhabitat types, forecasts and also draws attention to the danger that climate warming will probably enhance the spread of this detrimental C4 species.

The role of acetylcholine (ACh) as a signalling molecule in plants was investigated using a model system of Characeae cells. The effect of ACh on conductance of K+ channels in Nitella flexilis cells and on the action potential generation in Nitellopsis obtusa cells after H+-ATPase inhibition, where repolarization occurs after the opening of outward rectifying K+ channels, was investigated. Voltage-clamp method based on only one electrode impalement was used to evaluate the activity of separate potassium ion transport system at rest. We found that ACh at high concentrations (1 mM and 5 mM) activates K+ channels as the main membrane transport system at the resting state involved in electrogenesis of Characeaen membrane potential. We observed that ACh caused an increase in duration of AP repolarization of cells in K+ state when plasmalemma electrical characteristics are determined by large conductance K+ channels irrespective of whether AP were spontaneous or electrically evoked. These results indicate interference of ACh with electrical cellular signalling pathway in plants.

Fungi from the genus Ascochyta are generally facultative saprotrophs, which cause diseases in both monocots and dicots. Over 1 000 species belonging to this genus have been identified, 18 of which infect monocot plants from the family Poaceae. This study analyses the effects of temperature and light on the growth of selected fungi which infect monocots (A. agrostidis, A. avenae, A. brachypodii, A. desmazieri, A. digraphidis, A. ducis-aprutii, A. festucae, A. graminea, A. hordei, A. hordei var. americana, A. hordei var. europea, A. hordei var. hordei, A. melicae, A. phleina, A. skagwayensis, A. sorghi, A. stipae, A. zeicola), grown on three types of media; Potato Dextrose Agar (PDA), Coon’s agar (CN) and oatmeal agar (OMA). The fastest growth among the analyzed fungi at low temperatures was found in Ascochyta melicae, while at high temperatures it was A. zeicola. The fastest in vitro growth (average of all fungi) was observed on CN medium at 20°C (3.4 mm/day), while the lowest on OM medium at 5°C (1.0 mm/day). Radial mycelial growth in dark and the light conditions varied. On average, all isolates grew faster in the dark (3.1 mm/day) than in the light (1.9 mm/day). The greatest effect on the production of pycnidia was found for the isolates. Variation in growth and production of pycnidia depended on temperature, medium and lighting for fungi from the genus Ascochyta infecting monocots. Such variation indicates a potential occurrence of these fungi in different environments.

Breeding of new winter wheat cultivars with good heat tolerance requires better understanding of the genetic background of heat tolerance. In the present work the effect of heat stress on the 6th day after heading was investigated in a doubled haploid (DH) population arising from a cross between heat-sensitive (Plainsman V) and heat-tolerant (Mv Magma) cultivars. Averaged over the population, heat stress was found to result in a significant reduction in biomass, grain yield and grain number per plant, and in thousand-kernel weight (TKW) and harvest index. High temperature had the greatest effect on the grain yield, with a drop of 36.2% compared with the control. This could be attributed jointly to significant reductions in the TKW of the main ear and in the grain number of the side tillers. The relationship between the yield parameters was confirmed by the positive correlations obtained for the lines in the population. The diverse levels of heat tolerance in the different lines were confirmed by the significant differences in the reduction in the chlorophyll content (SPAD index) of the flag-leaves and in yield parameters. The changes in yield components in stress condition, however, can be still the most effective tools for heat stress evaluation.

The content of biologically active phenolic compounds (total polyphenols, tannins, flavonoids, and phenolic acids) were determined using spectrophotometry in four wild Croatian species of Daphne L. in the family Thymelaeaceae (Daphne alpina, D. cneorum, D. laureola, and D. mezereum). The concentration of total flavonoids (TF) was highest in the leaves of these Daphne species (0.12–0.51% dry herb weight, DW) whereas the content of other phenolic compounds analyzed were highest in the roots, including total polyphenols (TP; 2.71–19.03% DW), tannins (T; 1.14–7.39% DW), and total phenolic acids (TPA; 0.12–0.87% DW). D. alpina contained the highest amount of polyphenols, with the exception of flavonoids, where maximum concentrations were found in D. laureola. We also examined the antioxidant activity of leaf, stem, and root extracts. All extracts analyzed demonstrated high free radical scavenging activity with the highest concentration in the leaf extracts of D. alpina. Leaf extracts of D. cneorum showed the highest antioxidant activity in a β-carotene bleaching assay.

This work presents a study of the effect of foliar and root application of low concentrations (0.1–10 µM) of potassium naphthenate on the antioxidative status of cucumber (Cucumis sativus L.), assessed for both local and systemic organs. Changes in the contents of proline and glutathione indicate that the treatment of plants with potassium naphthenate can be characterized as a mild abiotic stress. The antioxidative system of cucumber plants is sensitive to such treatment, since organs directly exposed to the chemical showed a decrease in total antioxidant activities and an increase in peroxidation. In the organs that were not directly treated, an increase in the total antioxidative activity was observed only at the lowest naphthenate concentration while at higher concentrations this activity tended to decrease. As far as the activities of antioxidant enzymes (guaiacol peroxidase, superoxide dismutase, catalase) are concerned, the responses observed differed between enzymes for a given treatment, but showed similar trends within treated local and untreated systemic organs.

Thymus longicaulis C. Presl is a small aromatic perennial herb used as a traditional remedy for cold, flu and cough. Composition of the essential oil of T. longicaulis from Croatia and its in vitro antimicrobial activity against the most common respiratory pathogens were evaluated. The yield of essential oil obtained by hydrodistillation from aerial plant parts was 1.2%. According to the GC-MS analysis, a total of forty one compounds (99%) were identified. Thymol (46.3%), γ-terpinene (16.2%), thymyl methyl ether (11.4%), and p-cymene (9.4%) were the main components. Antimicrobial activity of the essential oil against six clinically isolated bacterial and yeast strains was determined using standard disc agar diffusion method and microdilution broth assay. The essential oil exhibited antimicrobial activity towards all tested respiratory pathogens. The most sensitive strains were Haemophilus influenzae and Streptococcus pneumoniae(MIC=0.78 mg/mL), while Staphylococcus aureus was the most resistant (MIC>25.00 mg/mL). Our results indicate that T. longicaulis essential oil could be effective against clinically relevant respiratory pathogens which have the ability to develop resistance to antimicrobial drugs.

The present study describes the total phenolic content, concentrations of flavonoids and in vitro antioxidant and antimicrobial activity of methanol extracts from Seseli pallasii Besser, S. libanotis (L.) Koch ssp. libanotis and S. libanotis (L.) Koch ssp. intermedium (Rupr.) P. W. Ball, growing wild in Serbia. The total phenolic content in the extracts was determined using Folin-Ciocalteu reagent and their amounts ranged between 84.04 to 87.52 mg GA (gallic acid)/g. The concentrations of flavonoids in the extracts varied from 4.75 to 19.37 mg Qu (quercetin)/g. Antioxidant activity was analyzed using DPPH reagent. Antioxidant activity ranged from 0.46 to 4.63 IC50 (mg/ml) and from 1.98 to 2.19 mg VitC (vitamin C)/g when tested with the DPPH and ABTS reagents, respectively, using BHA and VitC as controls. The antimicrobial activity of the extracts was investigated using a micro-well dilution assay for the most common human gastrointestinal pathogenic bacterial strains: Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 9027, Salmonella enteritidis ATCC 13076, Bacillus cereus ATCC 10876, Listeria monocytogenes ATCC15313, Staphylococcus aureus ATCC 25923 and Candida albicans ATCC 10231. This finding suggests that Seseli species may be considered as a natural source of antioxidants and antimicrobial agents.