Autologous stem cell therapy is the most promising alternative treatment in patients with chronic ischemic diseases, including ischemic heart disease and critical limb ischemia, which are characterized by poor prognosis related to serious impair of quality of life, high risk of cardiovascular events and mortality rates. However, one of the most serious shortcomings of stem cell transplantation are low survival after transplantation to the site of injury, as large number of stem cells are lost within 24 hours after delivery. Multiple studies suggest that combination of lipid-lowering drugs, statins, and stem cell transplantation might improve therapeutic efficacy in regenerative medicine. Statins are inhibitors of HMG-CoA reductase and belong to recommended therapy in all patients suffering from critical limb ischemia. Statins possess non-lipid effects which involve improvement of endothelial function, decrease of vascular inflammation and oxidative stress, anti-cancer and stem cell modulation capacities. These non-lipid effects are explained by inhibition of mevalonate synthesis via blocking isoprenoid intermediates synthesis, such as farnesylpyrophospate and geranylgeranylpyrophospate and result in modulation of the PI3K/Akt pathway. Moreover, statin-mediated microRNA regulation may contribute to the pleiotropic functions. MicroRNA interplay in gene regulatory network of IGF/Akt pathway may be of special significance for the treatment of critical limb ischemia. We assume further studies are needed for detailed analysis of statin interactions with microRNA at the molecular level and their link to PI3K/Akt and IGF/Akt pathway in stem cells, which are currently the most promising treatment strategy used in chronic ischemic diseases.
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
To investigate the transmission of species of Cryptosporidium Tyzzer, 1907 in Timis County, Romania, 48 isolates of Cryptosporidium coccidia from 11 children, 29 calves and eight pigs were characterised by molecular analysis of two loci (SSU rRNA and 60-kDa glycoprotein gene). Overall, 22 isolates were amplified and sequence analyses revealed that all isolates were Cryptosporidium parvum Tyzzer, 1912. Two subtype families were identified, IIa and IId. Subtype IIdA22G1 (n = 4) was the single C. parvum subtype found in children. Subtypes found in calves included IIdA27G1 (n = 8), a novel subtype, IIdA25G1 (n = 5), IIdA22G1 (n = 2), IIdA21G1a (n = 1), and IIaA16G1R1 (n = 1). Subtype IIdA26G1 was found in a pig. These results were significantly different from previous Romanian reports, as the five subtypes of family IId identified in this study were never identified previously in this country. Thus, cattle may be a source of Cryptosporidium infections for humans and the transmission dynamics of C. parvum in Romania is more complex than previously believed., Patrícia Manuela Vieira, Narcisa Mederle, Maria Luísa Lobo, Kálmán Imre, Ovidiu Mederle, Lihua Xiao, Gheorghe Darabus, Olga Matos., 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.
Exorista sorbillans, the uzi fly, is a serious tachinid pest of silkworm and is present in all silk producing areas of Asia. Assuming that E.sorbillans was accidentally transported from West Bengal to southern states of India, its population genetic structure was studied using 13 ISSR, 3 RAPD, two sets of universal primers and two sets of primers designed from a lepidopteran repeat sequence. Statistical analyses of DNA markers revealed significant genetic variability between the E. sorbillans populations from 4 different geographic locations (within 400 km of one another) in the southern states and the one from West Bengal (Murshidabad). Multivariate and discriminant function analyses indicate that the E. sorbillans from south India has diverged from the original gene pool of West Bengal and is suitable for studying the microevolution of adaptation to the conditions prevailing in the different cocoon producing areas in India.
Abbreviations used. GP = geographic population; ISSR = Inter Simple Sequence Repeat; PCR = Polymerase Chain Reaction; RAPD = Random Amplified Polymorphic DNA; SPSS = Statistical Package for Social Sciences; UBC = University of British Columbia; UNIV = Universal.
The ribosomal protein S6 kinase (S6K) plays a pivotal role in developmental processes and cell survival by participating in protein synthesis relevant signaling pathways. In the present study, an S6K gene (AccS6K-p70) was isolated and characterized from the Chinese honeybee, Apis cerana cerana (Fabricius) (Hymenoptera: Apidae), an important economic insect in the agricultural industry. The cDNA of AccS6K-p70 was 1683 bp in length and predicted to encode a protein of 467 amino acid residues. Sequence and structure analysis showed that there was a conserved catalytic domain in AccS6K-p70, whilst a phosphorylation site was found in the conserved part of the catalytic domain. Development relevant transcription factor binding sites found in the 5’-flanking region of AccS6K-p70 suggest that AccS6K-p70 might be involved in A. c. cerana development. Furthermore, quantitative PCR revealed that the expression levels of AccS6K-p70 were higher in head and thorax than in other tissues. The AccS6K-p70 was highly expressed in both larvae and adults compared with that in pupae, whilst expression of the gene was significantly down-regulated by hydrogen peroxide (H2O2) (although initially and slightly increased by it) and pyriproxyfen (a juvenile hormone analogue insecticide) stresses. These results suggest that AccS6K-p70 may play critical roles in developmental processes and cell survival in A. c. cerana, whilst both oxidative stress and pyriproxyfen may impair S6K-p70 mediated developmental processes by down-regulation of AccS6K-p70 expression., Yingqi Cai ... [et al.]., and Obsahuje seznam literatury
Insect cellular immune reaction, which generally includes phagocytosis, encapsulation and nodule formation, is achieved by hemocytes circulating in insect haemolymph. The shift of hemocytes from the normal phase to the adhering phase is an important process in the cellular immune reaction, which includes the attachment of hemocytes to foreign surfaces or other hemocytes via adhesion factors. Neuroglian is one of the adhering factors associated with encapsulation in Manduca sexta and Drosophila melanogaster. Here we studied the localization of neuroglian (MsNrg) in Mythimna separata and its functional role in the cellular immune reaction. The distribution of MsNrg mRNA between hemocyte populations was determined using real-time quantitative reverse transcription PCR and in situ hybridization, which revealed that MsNrg was highly expressed in adhering hemocytes, especially in plasmatocytes. Unexpectedly, the transcript was observed as well in non-adhering hemocytes, implying neuroglian has a function in non-adhering hemocytes. Moreover, we showed that the amount of MsNrg mRNA was not changed by injections of either biotic or abiotic non-selves. Fewer latex beads were fully encapsulated by hemocytes in larvae treated with MsNrg double-stranded RNA than in control larvae, but their ability to achieve phagocytosis and nodule formation remained unchanged by the MsNrg knockdown. These results indicate that the function of neuroglian in the cellular immune reaction is conserved in D. melanogaster and lepidopteran species, and neuroglian in non-adhering hemocytes could possess unidentified function., Kakeru Yokoi, Yoshiaki Kato, Masahiro Suzuki, Ken Miura., and Obsahuje bibliografii