Transient receptor potential A1 (TRPA1) is an excitatory ion channel that functions as a cellular sensor, detecting a wide range of proalgesic agents such as environmental irritants an d endogenous products of inflammation and oxidative stress. Topical application of TRPA1 agonists produces an acute nociceptive response through peripheral release of neuropeptides, purines and other transmitters from activated sensory nerve endings. This, in turn, further regulates TRPA1 activity downstream of G-protein and phospholipase C -coupled signaling cascades. Despite the important physiological relevance of such regulation leading to nociceptor sensitization and consequent pain hypersensitivity, th e specific domains through which TRPA1 undergoes post -translational modifications that affect its activation properties are yet to be determined at a molecular level. This review aims at providing an account of our current knowledge on molecular basis of r egulation by neuronal inflammatory signaling pathways that converge on the TRPA1 channel protein and through modification of its specific residues influence the extent to which this channel may contribute to pain., A. Kádková, V. Synytsya, J. Krusek, L. Zímová, V. Vlachová., and Obsahuje bibliografii
a1_Two full-length cDNAs (SGrca1 and SGrca2) encoding ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activase (RCA) were cloned from a heterophyllous aquatic plant, Sagittaria graminea, using Rapid-Amplification of cDNA Ends (RACE). SGrca1 contains a 1,320 bp open reading frame encoding a protein of 440 amino acids, and SGrca2 is exactly identical to SGrca1 except for 330 bp missing in the middle of SGrca1. Sequence analysis of cDNA and genomic DNA indicated both two cDNAs were generated from a common gene via alternative splicing. The deduced amino acid sequence encoded by SGrca1 showed 75-82% identity with other RCAs from higher plants and showed high homology in three highly conserved motifs associated with ATP-binding sites. RT-PCR analysis suggested both SGrca1 and SGrca2 were expressed in green tissues. During a 14 h light/10 h dark photoperiod, both aerial and submerged leaves exhibited the similar expression pattern of SGrca1 and SGrca2 with SGrca1 as the dominant form, but the accumulation of both SGrca1 and SGrca2 mRNA was significantly inhibited in the submerged leaves., a2_Western blot analysis showed that both SGrca1 and SGrca2 had their translation products, the 43 kDa form and the 31 kDa form expressing in leaves. Interestingly, the aerial leaves expressed higher amount of the 43 kDa form compared with the 31 kDa form, while it was reversed in the submerged leaves. The results demonstrated that both environments regulated the RCA gene expression at both transcriptional and posttranscriptional level. In addition, co-immunoprecipitation assay revealed that the isolated Rubisco-RCA complex contained both the 43 and 31 kDa forms, and the proportion of the 31 kDa form was obviously enhanced in the submerged leaves. The results indicated that both the 43 kDa and 31 kDa forms were involved in Rubisco and RCA interaction and the increased incorporation of the 31 kDa form was associated with submerged photosynthetic environment., D. Wang, S. Z. Xie, J. Yang, Q. F. Wang., and Obsahuje bibliografii
Microsporidia are intracellular parasites of insects and other higher eukaryotes. The microsporidian Nosema philosamiae Talukdar, 1961 was isolated from the eri silkworm, Philosamia cynthia ricini Grote. In the present study, alpha- and beta-tubulin genes from N. philosamiae were characterized. The identity analysis of nucleotide and amino acid sequences indicated high similarity with species of Nosema Nägeli, 1857 sensu lato (nucleotide sequences, ≥ 96.0%; amino acid sequences, ≥ 99.0%). However, the tubulin genes of N. philosamiae share low sequence similarity with that of N. ceranae Fries, Feng, da Silva, Slemenda et Pieniazek, 1996 (strain BRL01) and a Nosema/Vairimorpha species. Phylogenies based on alpha-, beta- and combined alpha- plus beta-tubulin gene sequences showed that N. philosamiae, along with the true Nosema species, forms a separate clade with a high bootstrap value, with N. ceranae BRL01 forming a clade of its own. The results indicated that the alpha- and beta-tubulin sequences may be useful as a diagnostic tool to discriminate the true Nosema group from the Nosema/Vairimorpha group.
The chlorophyll a/b-binding protein (CAB) serves in both photosystems (PS), I and II, as a coordinator of antenna pigments in the light-harvesting complex (LHC). The CABs constitute abundant and important proteins in the thylakoid membrane of higher plants. In our study, five CAB genes, which contained full-length cDNA sequences from the 4-year-old ginseng leaves (Panax ginseng Meyer), were isolated and named PgCAB. Phylogenetic comparison of the members of the subfamily between ginseng and higher plants, including Arabidopsis, revealed that the putative functions of these ginseng CAB proteins were clustered into the different family of Arabidopsis CABs; two PgCABs in LHCII family and three PgCABs in LHCI family. The expression analysis of PgCABs consistently showed dark-dependent inhibition in leaves. Expression analysis during abiotic stress identified that PgCAB genes responded to heavy metal, salinity, chilling, and UV stresses differently, suggesting their specific function during photosynthesis. This is the first comprehensive study of the CAB gene family in P. ginseng., J. Silva, Y. J. Kim, J. Sukweenadhi, S. Rahimi, W. S. Kwon, D. C. Yang., and Seznam literatury
Zoonotic cutaneous leishmaniasis (ZCL) is an expanding disease and a public health issue in Iran. In the present study, rate of natural infection of rodent populations with Leishmania was investigated in six endemic foci including 28 villages in Golestan, Esfahan, Yazd, Fars, Khuzestan and Ilam provinces. A total of 593 rodents were captured and identified as Rhombomys opimus (n = 325), Meriones libycus (n = 171), Meriones persicus (n = 27), Tatera indica (n = 37), Nesokia indica (n = 12), Rattus rattus (n = 13) and Mus musculus (n = 8). Microscopic examinations of Giemsa-stained smears showed that 108 out of 593 (18.2%) rodents were infected with Leishmania spp., whereas infection of 186 out of 593 (31.4%) rodents with Leishmania was then confirmed by ITS1-PCR. The highest rate of infection was found in R. opimus (prevalence of 35%) and M. libycus (31%). Based on Restriction Fragment Length Polymorphism (RFLP), 145 (78%) of 186 samples detected as Leishmania DNA were identified as L. major, 8 (4%) samples as L. turanica and 33 (18%) as mixed infection (L. major and L. turanica). Samples from infected rodents were inoculated subcutaneously at tail base of BALB/c mice. In 35 of them, nodules and ulcers containing amastigotes appeared at the inoculation site. The samples prepared from infected rodents were cultured in NNN medium and only two samples were positive. Rhombomys opimus, M. libycus, M. persicus, T. indica and N. indica were confirmed as reservoir hosts of ZCL in the studied regions. Leishmania major infection was usually accompanied L. turanica in naturally infected gerbils (R. opimus and M. libycus) in Golestan, Esfahan and Fars provinces.
Autosomal dominant hypercholesterolemia (ADH), more known as familial hypercholesterolemia (FH), is a lipid metabolism disorder characterized by an elevation in low-density lipoprotein cholesterol (LDL-C) and increased risk for cardiovascular disea se. In this study, we assessed a spectrum of mutations causing ADH in 3914 unrelated Czech patients with clinical diagnosis of hypercholesterolemia. Samples have been collected within the framework of the MedPed project running in the Czech Republic since 1998. So far we have found 432 patients (11.0 %) with the APOB gene mutation p.(Arg3527Gln) and 864 patients (22.1 %) with the LDLR gene mutation. In 864 probands carrying the LDLR gene mutation, 182 unique allelic variants were detected. We have identified 14 patients homozygous for mutations in the LDLR or APOB genes. We performed function analyses of p.(Leu15Pro) and p.(Gly20Arg) sequence variations., L. Tichý, L. Fajkusová, P. Zapletalová, L. Schwarzová, M. Vrablík, T. Freiberger., and Obsahuje bibliografii
The contemporary state of the problem of molecular masers observed In regions of active star formation is briefly reviewed. The major landmarks of the history of their discovery, observations,
and interpretations are mentioned. OH masers observed In these regions are believed to be a product of disintegration of dense
molecular envelopes ("cocoons") surroundlng compact HlI regions of young OB-stars. They are local ized at, typically, =:10^17 cm from their parent star and have there densities = 10^6 cm, temperatures
=100 K, magnetic fields of the order of several mG. At least in some cases they seem to be falling on the star.
H2O (and, as an exception, Si0) masers are connected with still earlier stages of stellar evolutlon, when a very strong mass loss from the saár (up to =10^-3 - 10^-2 M /yr) takes place. H2O maser emission is produced by gas-dynamical (possibly MHD) interaction between the stellar wind and the surroundlng gas.The H2O sources may be localized much closer to their stars than the OH sources, - up to =10^14 - 10^15 cm, and they indicate much higher densities (=-10^11 cm^-3) and magnetic fields (up to several G), and higher temperatures ( = 100 - 1000 K). They reveal also rather llvely klnematics of the circumstellar gas (up to several hundred km/s).
CH3OH 1.3 cm low-gain masers in Orion with their observed dimensions and derived densities and temperatures may be gravitationally unstable and, with their posltion around a
pre-Trapezium young star cluster, may perfectly be a group of protostars. Much stronger CH3OH 2.5 cm masers, recently discovered in the direction of many compact HII regions, are dosely associated with the OH masers.
This review summarizes recent information on the role of calcium in the process of neuronal injury with special attention to the role of calcium stores in the endoplasmic reticulum (ER). Experimental results present evidence that ER is the site of complex processes such as calcium storage, synthesis and folding of proteins and cell response to stress. ER function is impaired in many acute and chronic diseases of the brain which in turn induce calcium store depletion and conserved stress responses. Understanding the mechanisms leading to ER dysfunction may lead to recognition of neuronal protection strategies., J. Lehotský, P. Kaplán, E. Babušíková, A. Strapková, R. Murín., and Obsahuje bibliografii
The Microsporidia are a group of obligate intracellular parasites, now thought to be derived fungi. Presented here is a comparative small subunit rDNA (ssrDNA) analysis of 125 species of Microsporidia (sequences obtained from GenBank). This analysis shows that groups or clades are formed based largely on habitat and host. This result is supported by comparative molecular analyses of the past decade, and indicates that structural and ultrastructural characters are unreliable for distinguishing among higher-level microsporidian taxa. Our findings indicate the presence of five major clades of Microsporidia which group according to habitat. We present three new classes of Microsporidia based on natural phylogenetic groupings as illustrated by the ssrDNA analysis: Aquasporidia, Marinosporidia and Terresporidia. The names of the proposed classes reflect the habitat of each group. The class Aquasporidia, found primarily in freshwater habitats, is a paraphyletic group consisting of three clades. The Marinosporidia are found in hosts of marine origin and the Terresporidia are primarily from terrestrial environments.
Summary The aim of the study was to characterize by molecular profiling two glomerular diseases: IgA nephropathy (IgAN) and focal segmental glomerulosclerosis (FSGS) and to identify potential molecular markers of IgAN and FSGS progression. The expressions of 90 immune-related genes were compared in biopsies of patients with IgAN (n=33), FSGS (n=17) and in controls (n=11) using RT-qPCR. To identify markers of disease progression, gene expression was compared between progressors and non-progressors in 1 year follow-up. The results were verified on validation cohort of patients with IgAN (n=8) and in controls (n=6) using laser-capture microdissection, that enables to analyze gene expression separately for glomeruli and interstitium. In comparison to controls, patients with both IgAN and FSGS, had lower expression of BAX (apoptotic molecule BCL2-associated protein) and HMOX-1 (heme oxygenase 1) and higher expression of SELP (selectin P). Furthermore, in IgAN higher expression of PTPRC (protein-tyrosine phosphatase, receptor-type C) and in FSGS higher expression of BCL2L1 (regulator of apoptosis BCL2-like 1) and IL18 compared to control was observed. Validation of differentially expressed genes between IgAN and controls on another cohort using laser-capture microdissection confirmed higher expression of PTPRC in glomeruli of patients with IgAN. The risk of progression in IgAN was associated with higher expression EDN1 (endothelin 1) (AUC=0.77) and FASLG (Fas ligand) (AUC=0.82) and lower expression of VEGF (vascular endothelial growth factor) (AUC=0.8) and in FSGS with lower expression of CCL19 (chemokine (C-C motif) ligand 19) (AUC=0.86). Higher expression of EDN1 and FASLG along with lower expression of VEGF in IgAN and lower expression of CCL19 in FSGS at the time of biopsy can help to identify patients at risk of future disease progression., I. Tycová, P. Hrubá, D. Maixnerová, E. Girmanová, P. Mrázová, L. Straňavová, R. Zachoval, M. Merta, J. Slatinská, M. Kollár, E. Honsová, V. Tesař, O. Viklický., and Seznam literatury