Band 5, Heft 3

Mitochondrial toxins represent an interesting group of neurotoxicants related both to causation and modelling of central nervous damage. 3-nitropropionic acid (3NP), a neurotoxin of herbal and microfungal origin, inhibits succinate dehydrogenase leading thereby to various biochemical and morphological alterations in the brain. Experimental animals treated by 3NP are used to model Huntington’s disease. Manganese, often present in occupational settings and as environmental pollutant, inhibits complex II and III of the mitochondria and is known to cause Parkinson-like CNS damage. In this work, rats were administered a single acute dose of Mn (50 mg Mn2+/kg body weight) or 3-NP (20 mg/kg b.w.) and the alterations of the somatosensory cortical evoked potential elicited by stimulation of the whisker pad and the tail base were observed, together with the changes of the action potential in the tail nerve. Latency and amplitude of the two cortical responses changed in parallel, while those of the tail nerve response remained more or less unaltered. The two mitochondrial toxins studied seem to exert their action centrally, primarily on synaptic transmission, rather than peripherally. Recording of evoked activity could be used to follow-up the nervous system effects of mitochondrial toxins, but it requires further investigation.

The effects of amidated pectin and psyllium on serum, hepatic and faecal cholesterol concentration were compared in female rats fed diets supplemented with palm fat and cholesterol at 50 and 10 g/kg, respectively. Control rats were fed a diet supplemented with cellulose at 60 g/kg. In treated rats, cellulose was replaced with either amidated pectin or psyllium. Amidated pectin and psyllium intake significantly decreased serum cholesterol from 3.41 μmol/ml (control) to 1.68 and 2.04 μmol/ml, respectively, and hepatic cholesterol from 31.9 μmol/g (control) to 7.2 and 9.0 μmol/g, respectively. Histology with lipid-staining Sudan Black B revealed that liver tissue from control rats was infiltrated with lipids, but staining was absent in livers of treated rats. No hepatic pathophysiology was apparent in treated rats. Amidated pectin and psyllium intake significantly increased faecal fat content. Faecal cholesterol content was significantly increased in rats that were fed amidated pectin, and non-significantly increased in rats that were fed psyllium. Body weight and food intake did not differ among treatment groups. In conclusion, amidated pectin, a novel sequestrant of sterols, demonstrated a similar effect on rat serum and hepatic cholesterol concentration to psyllium, which is a well-established hypocholesterolaemic agent.

The present study was designed to explore the mechanism of action of walnut (the seed of Juglans regia) leaf and ridge on hepatic glucose metabolism in diabetic mice. Experimental diabetes was induced by intravenous administration of streptozotocin (60 mg/kg)and confirmed with an increase of blood glucose, 90–100% of the control, 72 hours later. Isolated extracts from walnut leaf and ridges were administered in a single effective dose of 400 mg/kg orally. Firstly, blood glucose was determined every 1 hour until 5 hours post administration of extracts. In the second experiment, the liver was surgically removed, 2 hours post treatment of diabetic animals with extracts, homogenized and used for measurement of key enzymes of glycogenolysis (glycogen phosphorylase, GP) and gluconeogenesis (phosphoenolpyruvate carboxykinase, PEPCK). Treatment by both leaf and ridge extracts decreased blood glucose and liver PEPCK activity and increased blood insulin and liver GP activity. It is concluded that walnut is able to lower blood glucose through inhibition of hepatic gluconeogenesis and secretion of pancreatic insulin.

Polyclonal antibodies, as well as monoclonal antibodies are efficacious in providing protective immunity against Francisella tularensis. This study demonstrates the application of phage display libraries for the construction of monoclonal antibodies against F. tularensis. Novel single-chain fragment variable (scFv) antibodies were generated against a whole bacterial lysate of F. tularensis live vaccine strain using the human single fold scFv libraries I (Tomlinson I + J). A total of 20 clones reacted with the bacterial cell lysate. Further, the library contains two clones responsive to recombinant lipoprotein FTT1103Δsignal (F. tularensis subsp. tularensis Schu S4), which was constructed without a signal sequence. These positively-binding scFvs were evaluated by scFv-phage enzyme-linked immunosorbent assay (ELISA). Then, positive scFvs were expressed in a soluble form in Escherichia coli HB2151 and tested for positive scFvs by using scFv-ELISA.

In the last decade, infections caused by Candida glabrata have become more serious, particularly due to its decreased susceptibility to azole derivatives and its ability to form biofilm. Here we studied the resistance profile of 42 C. glabrata clinical isolates to different azoles, amphotericin B and echinocandins. This work was also focused on the ability to form biofilm which plays a role in the development of antifungal resistance. The minimal inhibitory concentration testing to antifungal agents was performed according to the CLSI (Clinical and Laboratory Standards Institute) M27-A3 protocol. Quantification of biofilm was done by XTT reduction assay. All C. glabrata clinical isolates were resistant to itraconazole and sixteen also showed resistance to fluconazole. All isolates remained susceptible to voriconazole. Amphotericin B was efficient in a concentration range of 0.125–1 mg/L. The most effective antifungal agents were micafungin and caspofungin with the MIC100 values of ≤0.0313–0.125 mg/L. Low concentrations of these agents reduced biofilm formation as well. Our results show that resistance of different C. glabrata strains is azole specific and therefore a single azole resistance cannot be assumed to indicate general azole resistance. Echinocandins proved to have very high efficacy against clinical C. glabrata strains including those with ability to form biofilm.

The Major Histocompatibility Complex (MHC) is a large gene family that is found in most vertebrates and has an important influence on body odour preference and mate selection in animals. In this research we found, that human leukocyte antigen (HLA) phenotype is strongly connected with the strength and pleasantness of perceived odour of selected chemical compounds found in sweat. Among different chemical classes of compounds tested, the esters of fatty acids such as methyl undecanoate, methyl decanoate, methyl nonanoate, methyl octanoate and methyl hexanoate show strongest connection to HLA. On the other hand, our experiment did not confirm the connection of MHC to the perceived strength and pleasantness of body odour.

The antioxidant properties of methanol extracts of Lady’s Bedstraw (Galium verum L., Rubiaceae) herb from two different localities in Serbia were evaluated. Antioxidant activity was assessed in four different model systems. Free radical scavenging capacity (RSC) was examined by measuring the scavenging activity of extracts on 2,2-diphenyl-1-pycrylhydrazil (DPPH) and hydroxyl radical (OH), as well as on hydrogen peroxide. In addition, the protective effects of lipid peroxidation (LP) in corn oil were evaluated by the TBA-assay using the Fe2+/ascorbate system of induction. The amount of dried extract, the content of total phenolics, flavonoids and chlorophylls was also determined. Extracts from both locations expressed very strong scavenger activity, reducing the DPPH⊙ (IC50=3.10 µg/mland 8.04 µg/ml) and OH radical formation (IC50=0.05 µg/ml and 0.54 µg/ml) and neutralising H2O2 (IC50=4.98 µg/ml and 3.80 µg/ml), in a dose dependant manner. Also, examined extracts showed notable inhibition of LP (IC50=11.69 µg/ml and 19.47 µg/ml). The observed differences in antioxidant activity could be partially explained by the levels of phenolics (2.44–4.65 mg and 4.57–5.16 mg gallic acid equivalents/g dry extract), flavonoids (6.38–10.70 µg and 15.56–17.96 µg quercetin equivalents/g dry extract) and chlorophylls in the investigated Lady’s Bedstraw extracts.

Antioxidant activity of the aerial part and bulbs of Allium paradoxum was investigated by eight in vitro assay systems. Extracts showed good antioxidant activity. IC50 for 1,1-diphenyl-2-picryl hydrazyl radical-scavenging activity was 890.9±43.2 and 984.9±33.5 µg/ml for the aerial part and bulbs, respectively. The aerial parts have better reducing power than bulb extracts but not comparable with Vitamin C (P>0.001). Extracts showed weak Fe2+ chelating ability, the IC50 being 959±47 and 530±24 µg/ml for bulbs and aerial parts, respectively. Both tested extracts exhibited good hydrogen peroxide scavenging in a concentration dependent manner. They exhibited good antioxidant activity against the hemoglobin-induced linoleic acid system that was comparable with vitamin C (P>0.01). They showed good activity against cumene hydro peroxide induced hemolysis in RBCs. In addition, they possessed antihemolytic activity. The extract from aerial parts had significantly higher total phenol and flavonoid content than did bulbs. Amounts of eight elements (Cu, Mn, Zn, Fe, Ni, Pb, Cd and Cr) were also determined in the bulb and aerial part using atomic absorption spectroscopy. They contained higher Fe and Mn contents than other elements.

Differences in essential oil composition of wild Achillea millefolium L., collected at five habitats in Lithuania, where plants with pink (f. rosea) and white (f. millefolium) flowers grow together, were reported. For the first time, oils of different plant organs (inflorescences and leaves) of both forms from every population were analysed in detail. Chemical analysis was performed by GC and GC-MS. The most predominant constituents of the oils were nerolidol (9.4–31.9%, in 11 out of 20 samples), caryophyllene oxide (8.4–23.0%, 4 leaf oils), ß-pinene (8.0–15.2%, 2 samples), eudesmol (11.8–15.8%, 2 leaf oils) and 1,8-cineole (11.9%, one inflorescence oil). Domination of nerolidol was mostly characteristic of A. millefolium f. rosea essential oils (in 8 out of 10 oils). The inflorescences biosynthesised markedly larger amounts of nerolidol and ß-pinene than those of the leaves. An opposite correlation was observed for caryophyllene oxide and eudesmol. Chamazulene (≤2.7%) was determined only in six oils. The 65 identified constituents made up 75.4–96.5% of the oils.

Galactoglucomannan oligosaccharides (GGMOs, d.p. 4–8 and fractions d.p. 3, 4, 5, 6–7), used in culture media for spruce protoplasts derived from callus cells showed a pleiotropic effect. They influenced both, quality and quantity of extracellular proteins in regenerating protoplasts. GGMOs d.p. 4, 5 at pH 6.0 and the mixture of d.p. 4–8 (pH 3.8 and 6.0) after 48 h of culture increased the amount of extra- and intracellular proteins and the viability of protoplasts. The most significant effect on protoplasts viability in the presence of GGMOs d.p. 4–8 without the growth hormone supplementation at pH 3.8, and with lower efficiency in the presence of NAA (1 mg/l) has been observed. The most significant differences were observed in the molecular mass intervals Mr∼17–21 kDa, 25–30 kDa, and 45–60 kDa. After 24 h of culture extracellular acid proteins with Mr∼30; 41; 68; and 90 in media supplemented with GGMOs d.p. 3, 4, 5, 6–7 (pH 6.0) with or without NAA were identified to belong to the group of β-1,3-glucanases. Extracellular proteins p27.5; 30; 41; 50; 52 and 90 exhibited chitinase activity after 24 h of protoplast cultivation. GGMOs probably fulfil a protective role in this process of spruce protoplast regeneration.

The effects of Aporrectodea caliginosa earthworms on both carbon dioxide (CO2) accumulation in and emissions from soil, as well as the simultaneous impact of earthworms on soil microbiological properties were investigated in a microcosm experiment carried out over 5.5 months. Concentration of CO2 in soil air was greater at a depth of 15 cm when compared with a depth of 5 cm, but varied during the season both in control and earthworm-inhabited chambers. Peaks of CO2 concentrations at both depths occurred in both treatments during August, approximately 80 days after the experiment started. Generally, the presence of earthworms increased the CO2 concentration at 15-cm depth. Larger CO2 emissions were consistently recorded in conjunction with higher amounts of CO2 in soil air when chambers were inhabited by earthworms. The total CO2 emissions during the experimental period covering 161 days were estimated at 118 g CO2-C m−2 and 99 g CO2-C m−2 from chambers with and without earthworms respectively. Moreover, the presence of earthworms increased microbial biomass in the centre and at the bottom of chambers, and enhanced both dehydrogenase activity and nitrifying enzyme activity in the soils. We suggest that the effect of earthworms on both the enhanced soil accumulation of CO2 as well as emissions of CO2 was mostly indirect, due to the impacts of earthworms on soil microbial community.

The paper deals with the health status, production and structure of autochthonous beech stands in the eastern part of the Krkonoše Mts. (Czech Republic). The region was affected by increased air pollution load due to long-range SO2 transmission from the late 1970s to the early 1990s. In 1980, five research plots were established in long unmanaged beech stands at an optimum stage. Tree coordinates and initial biometrical measurements were made in 1980 and repeated in 2005. The defoliation of individual trees was monitored every year (ICP Forests principles). The actual stand development is compared with the model development worked out by the Sibyla growth simulator in variants with a model of natural mortality and with entering a real mortality of trees. Differences between the stand characteristics of actual and simulated development of forest stands suggest some production losses. The reason to the losses can be seen in the long-term impact of air pollution in the first decade of monitoring and its after-effects on soil quality and worsening health condition of stands. The anticipated acceleration in the onset of the phase of stand disintegration under the influence of air pollution load was not demonstrated.

Changes in circadian rhythms of dROMs, Oxy-ads and SHp during reproductive stages were studied in Comisana ewes. Twelve ewes were divided in two equal groups. The experimental group consisted of ewes undergoing gestation and lactation following artificial insemination and the control group consisted of non-pregnant ewes. Blood samples were collected every 3 h over a 24 h period, 20 days before insemination, on days 100 and 140 of pregnancy, on days 10, 30 and 200 post-partum and during the dry period. In the control group, blood samples were collected on the same days and with the same procedures as those used for the experimental group. A significant effect of time on all parameters studied was observed in the experimental group. Daily rhythms of the parameters studied were observed in the control group in all experimental conditions, and in the experimental group during pre-pregnancy and dry periods. We conclude that the reproductive status of sheep affects oxidative stress markers in blood and their circadian rhythms.

We present data on the female reproductive traits of the Balkan wall lizard in the Deliblato Sand, a large continental sandland in the Pannonian area in the northwestern periphery of the species range. The clutch and egg characteristics of the population were investigated on the basis of clutches laid in laboratory conditions by gravid females captured in one locality. Balkan wall lizards produced at least two clutches in a breeding season. Individual females laid clutches of commonly two (range 1–4) eggs. The female body size had no effect on clutch and egg size. There was no trade-off between egg size and clutch size.

As a result of investigations over type material from a collection of the Hungarian Natural History Museum, Budapest, a new synonym and two new combinations in the Torymidae family (Hymenoptera: Chalcidoidea) are proposed: Eridontomerus illiesi Szelényi, 1981 as a junior synonym of Cryptopristus caliginosus (Walker, 1833); Eridontomerus unidentatus Szelényi, 1973 is transferred to the genus Pseudotorymus Masi, 1921; and Idiomacromerus obscuripennis (Szelényi, 1973) is transferred to the genus Torymoides Walker, 1871.