The influence of CRF and alpha-helical CRF(9-41) on rat fear responses, c-Fos and CRF expression, and concentration of amino acids in brain structures

In the present study we have examined the influence of intracerebroventricullary administered CRF, and a non-selective CRF receptor antagonist, α-helical CRF_(9–41), on rat conditioned fear response, serum corticosterone, c-Fos and CRF expression, and concentration of amino acids (in vitro), in several brain structures. Pretreatment of rats with CRF in a dose of 1μg/rat, enhanced rat-freezing response, and further increased conditioned fear-elevated concentration of serum corticosterone. Moreover, exogenous CRF increased aversive context-induced expression of c-Fos in the parvocellular neurons of the paraventricular hypothalamic nucleus (pPVN), CA1 area of the hippocampus, and M1 area of the frontal cortex. A different pattern of behavioral and biochemical changes was present after pre-test administration of α-helical CRF_(9–41) (10μg/rat): a decrease in rat fear response and serum corticosterone concentration; an attenuation of fear-induced c-Fos expression in the dentate gyrus, CA1, Cg1, Cg2, and M1 areas of the frontal cortex; a complete reversal of the rise in the number of CRF immunoreactive complexes in the M2 cortical area, induced by conditioned fear. Moreover, α-helical CRF_(9–41) increased the concentration of GABA in the amygdala of fear-conditioned rats. Altogether, the present data confirm and extend previous data on the integrative role of CRF in the central, anxiety-related, behavioral and biochemical processes. The obtained results underline also the role of frontal cortex and amygdala in mediating the effects of CRF on the conditioned fear response.     

Recent drastic changes in the gammarid fauna (Crustacea, Amphipoda) of the Vistula River deltaic system in Poland caused by alien invaders

During the last decade of 20th century, the nonindigenous gammarid species Gammarus tigrinus, Dikerogammarus haemobaphes, Pontogammarus robustoides and Obesogammarus crassus invaded the lower Vistula River and its deltaic, partly brackish regions. G. tigrinus, an oligohaline North‐American species, was introduced to western Europe in the 1950s; the remaining three species are oligohaline/freshwater Ponto‐Caspian species. All these species are now invading central and western Europe using the network of man‐made canals connecting different European river systems. In the Vistula River, the native European freshwater gammarid species Gammarus pulex and G. varsoviensis were replaced in the 1920s by the Ponto‐Caspian Chaetogammarus ischnus (syn. Echinogammarus ischnus), which in turn has been outnumbered by the more recent invasions of D. haemobaphes and P. robustoides. In brackish waters, the native Atlantic‐boreal species Gammarus zaddachi and Gammarus duebeni are replaced or at least outnumbered by G. tigrinus, P. robustoides and O. crassus. Possible invasion routes are discussed.     

ESBL-positive and ESBL-negative Klebsiella pneumoniae and Klebsiella oxytoca strains--occurrence in clinical specimens and susceptibility to selected antibiotics

The aim of this study was to evaluated susceptibility to selected antibiotics and occurrence in clinical materials ESBL-positive and ESBL-negative K. pneumoniae and K. oxytoca strains. The susceptibility to selected antibiotics were tested by disk-diffusion method. None of the analysed strains was resistant to imipenem. Among non beta-lactam antibiotics thwe most strains were susceptible to fluoroquinolones. The most frequently K. pneumoniae strains were isolated from urine, K. oxytoca trains were isolated from wounds.     

Functional and structural basis for a bacteriophage homolog of human RAD52

In eukaryotes, homologous recombination proteins such as RAD51 and RAD52 play crucial roles in DNA repair and genome stability. Human RAD52 is a member of a large single-strand annealing protein (SSAP) family [1] and stimulates Rad51-dependent recombination [2, 3]. In prokaryotes and phages, it has been difficult to establish the presence of RAD52 homologs with conserved sequences. Putative SSAPs were recently found in several phages that infect strains of Lactococcus lactis[4]. One of these SSAPs was identified as Sak and was found in the virulent L. lactis phage ul36, which belongs to the Siphoviridae family [4, 5]. In this study, we show that Sak is homologous to the N terminus of human RAD52. Purified Sak binds single-stranded DNA (ssDNA) preferentially over double-stranded DNA (dsDNA) and promotes the renaturation of long complementary ssDNAs. Sak also binds RecA and stimulates homologous recombination reactions. Mutations shown to modulate RAD52 DNA binding [6] affect Sak similarly. Remarkably, electron-microscopic reconstruction of Sak reveals an undecameric (11) subunit ring, similar to the crystal structure of the N-terminal fragment of human RAD52 [7, 8]. For the first time, we propose a viral homolog of RAD52 at the amino acid, phylogenic, functional, and structural levels.     

Ligand-independent exhaustion of killer immunoglobulin-like receptor-positive CD8+ T cells in human immunodeficiency virus type 1 infection

Virus-specific CD8⁺ T cells play a central role in the control of viral infections, including human immunodeficiency virus type 1 (HIV-1) infection. However, despite the presence of strong and broad HIV-specific CD8⁺ T-cell responses in chronic HIV-1 infection, these cells progressively lose critical effector functions and fail to clear the infection. Mounting evidence suggests that the upregulation of several inhibitory regulatory receptors on the surface of CD8⁺ T cells during HIV-1 infection may contribute directly to the impairment of T-cell function. Here, we investigated the role of killer immunoglobulin receptors (KIR), which are expressed on NK cells and on CD8⁺ T cells, in regulating CD8⁺ T-cell function in HIV-1 infection. KIR expression was progressively upregulated on CD8⁺ T cells during HIV-1 infection and correlated with the level of viral replication. Expression of KIR was associated with a profound inhibition of cytokine secretion, degranulation, proliferation, and activation by CD8⁺ T cells following stimulation with T-cell receptor (TCR)-dependent stimuli. In contrast, KIR⁺ CD8⁺ T cells responded potently to TCR-independent stimulation, demonstrating that these cells are functionally competent. KIR-associated suppression of CD8⁺ T-cell function was independent of ligand engagement, suggesting that these regulatory receptors may constitutively repress TCR activation. This ligand-independent repression of TCR activation of KIR⁺ CD8⁺ T cells may represent a significant barrier to therapeutic interventions aimed at improving the quality of the HIV-specific CD8⁺ T-cell response in infected individuals.     

The effect of biochanin A on the chlorophylls and carotenoids content in the alga <Emphasis Type="Italic">Chlorella vulgaris</Emphasis> Beijerinck

The present study was undertaken to determine the influence of biochanin A, isoflavone characterised by estrogenic activity, upon the content of chlorophylls and carotenoids in the cells of green alga Chlorella vulgaris Beijerinck (Chlorophyceae). On the 6^th day of cultivation under the influence of 10⁻⁶ M biochanin A exerted the greatest biological activity and the most stimulating effect on the analysed parameters: growth of the alga expressed by the cells number and the content of photosynthetic pigments in them. The total content of carotenoids was stimulated on the 6^th day of experiment in the range of 197 % but during the 9^th day only in 179 % in comparision with the control group (100 %). At the same time content of carotenes increased to the level of 123 – 119 % and xanthophylls to 208 – 178 %. Among the carotenes, β-carotene was characterised with the 3.7 times higher content in regard to the content of α-carotene on the 6^th day of cultivation and during the 9^th day — the 5.7 times domination. The content of xanthophylls that contain two atoms of oxygene in molecule (oxygen — poor xanthophylls) was intensively stimulated in the range of 224 %. Moreover, the oxygen — rich xanthophylls content reached the value 179 % when compared to the control. The greatest stimulation of the content of chlorophylls and its isomers was observed during the 3^rd day of cultivation of Chlorella vulgaris when it rose up to 166 % and to 156 % on the 6^th day. The content of chlorophyll b and its isomers was stimulated in 181 % on the 6^th day of culture and 155 % during the 9^th day of algal culture. The evidence on the stimulating effect of biochanin A as the main representative of isoflavones on the growth and content photosynthetic pigments in eucaryotic alga C. vulgaris was demonstrated in these studies.     

Substrate specificity of rat brain ceramidase.

In this study, the substrate specificity of a newly identified rat brain ceramidase (CDase) was investigated. To this end, the major functional groups and stereochemistry of ceramide (Cer) were evaluated for their influence on the hydrolysis of substrate by this CDase. The results showed that, of the four possible stereoisomers of Cer, only the natural d-e-C(18)-Cer isomer was used as substrate (K(m) of 1.1 mol% and V(max) of 5 micromol/min/mg). Removal of the 4-5 trans double bond to generate dihydroceramide decreased the affinity of the enzyme toward its substrate by around 90%, whereas changing the configuration of the double bond from the natural trans configuration into cis or introduction of a hydroxyl group (phytoceramide) resulted in loss of hydrolysis. Shortening the chain length of the sphingosine backbone resulted in decreased affinity. Methylation of either the primary or the secondary hydroxyl groups resulted in loss of activity. Results also indicated that Cer species that harbor long saturated or monounsaturated fatty acyl chains are preferred substrates of the enzyme. alpha-Hydroxylated Cer demonstrated considerably higher affinity, indicating a preference of the enzyme to those Cer molecular species. These results disclose a very high specificity of nonlysosomal CDase for its substrate, Cer.     

<Emphasis Type="Italic">Raoultella</Emphasis> spp.—clinical significance,infections and susceptibility to antibiotics

The genus Raoultella belongs to the family of Enterobacteriaceae. Raoultella spp. are Gram-negative, aerobic, non-motile rods. This genus can be distinguished from the genus Klebsiella, in that genus use histamine as the only source of carbon in the medium. Also, Raoultella grow at 4 °C and do not produce gas from lactose at 44.5 °C. Raoultella sp. is known to inhabit natural environments (water, soil, plants). The reservoir of Raoultella is the gastrointestinal tract and upper respiratory tract. Raoultella spp. are opportunistic bacteria, which usually cause infections of the biliary tract, pneumonia and bacteraemia in oncologic and with lower immunity patients. Raoultella planticola and Raoultella ornithinolytica are the most frequently encountered human pathogens among the genus Raoultella. In this review, the current knowledge on Raoultella infections is summarized.