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31.
RuvB family of protein contains two similar kinds of proteins i.e. RuvB1 and RuvB2 from yeast to human. These proteins belong to the AAA + class of proteins and are critical components of several multiprotein complexes involved in diverse cellular activities. There are two RuvB proteins annotated in the Plasmodium database but the identification of the third protein recently by our lab has raised the question why Plasmodium falciparum contains three RuvB proteins instead of two. Hence the biochemical characterizations of these proteins have become essential to understand the role of these proteins in the malaria parasite. Recently we have reported the characterization of the recombinant PfRuvB3, which contains ATPase activity but lacks DNA helicase activity. In the present study we report the phylogenetic analysis and detailed biochemical characterization of one of the other RuvB homologue RuvB1 from P. falciparum. PfRuvB1 shows considerable homology with human as well as yeast RuvB1 and contains Walker motif A and Walker motif B. The activity analysis of this protein revealed that PfRuvB1 is an ATPase and this activity increased significantly in the presence of ss-DNA. PfRuvB1 also contains DNA helicase activity and translocates preferentially in 5′ to 3′ direction. In vivo investigation of PfRuvB1 revealed that it is constitutively expressed during all the stages of intraerythrocytic cycle of P. falciparum and localizes mainly to the nucleus. These studies will make important contribution in understanding the role of RuvB protein in P. falciparum. 相似文献
32.
Luiz Claudio Pereira Ribeiro Cassia Cristina Alves Gon?alves Carla Maria Sena Andrade Slater Silvia Maia Farias de Carvalho Marzia Puccioni-Sohler 《Memórias do Instituto Oswaldo Cruz》2013,108(6):730-734
Intrathecal synthesis of human T-lymphotropic virus type 1 (HTLV-1) antibodies
(Abs) represents conclusive evidence of a specific immune response in the
central nervous system of HTLV-1 associated myelopathy/tropical spastic
paraparesis (HAM/TSP) patients. Western blotting (WB) for HTLV Abs in serum is a
confirmatory test for HTLV-1 infection. The aim of this study was to standardise
the Western blot to demonstrate the intrathecal pattern of Abs against HTLV-1
proteins in HAM/TSP patients. Paired cerebrospinal fluid (CSF) and serum samples
were selected from 20 patients with definite HAM/TSP, 19 HTLV-1 seronegative
patients and two HTLV-1 patients without definite HAM/TSP. The presence of
reactive bands of greater intensity in the CSF compared to serum (or bands in
only the CSF) indicated the intrathecal synthesis of anti-HTLV-1 Abs. All
definite HAM/TSP patients presented with an intrathecal synthesis of anti-HTLV-1
Abs; these Abs were not detected in the control patients. The most frequent
intrathecal targets of anti-HTLV-1 Abs were GD21, rgp46-I and p24 and, to a
lesser extent, p19, p26, p28, p32, p36, p53 gp21 and gp46. The intrathecal
immune response against env (GD21 and rgp46-I) and
gag (p24) proteins represents the most important humoral
pattern in HAM/TSP. This response may be used as a diagnostic marker,
considering the frequent association of intrathecal anti-HTLV-1 Ab synthesis
with HAM/TSP and the pathogenesis of this neurological disease. 相似文献
33.
Adriana Simionescu-Bankston Giovanna Leoni Yanru Wang Peter P. Pham Arivudainambi Ramalingam James B. DuHadaway Victor Faundez Asma Nusrat George C. Prendergast Grace K. Pavlath 《Developmental biology》2013
Actin dynamics are necessary at multiple steps in the formation of multinucleated muscle cells. BAR domain proteins can regulate actin dynamics in several cell types, but have been little studied in skeletal muscle. Here, we identify novel functions for the N-BAR domain protein, Bridging integrator 3 (Bin3), during myogenesis in mice. Bin3 plays an important role in regulating myofiber size in vitro and in vivo. During early myogenesis, Bin3 promotes migration of differentiated muscle cells, where it colocalizes with F-actin in lamellipodia. In addition, Bin3 forms a complex with Rac1 and Cdc42, Rho GTPases involved in actin polymerization, which are known to be essential for myotube formation. Importantly, a Bin3-dependent pathway is a major regulator of Rac1 and Cdc42 activity in differentiated muscle cells. Overall, these data classify N-BAR domain proteins as novel regulators of actin-dependent processes in myogenesis, and further implicate BAR domain proteins in muscle growth and repair. 相似文献
34.
ABSTRACT:?Xylanases are hydrolases depolymerizing the plant cell wall component xylan, the second most abundant polysaccharide. The molecular structure and hydrolytic pattern of xylanases have been reported extensively and the mechanism of hydrolysis has also been proposed. There are several models for the gene regulation of which this article could add to the wealth of knowledge. Future work on the application of these enzymes in the paper and pulp, food industry, in environmental science, that is, bio-fueling, effluent treatment, and agro-waste treatment, etc. require a complete understanding of the functional and genetic significance of the xylanases. However, the thrust area has been identified as the paper and pulp industry. The major problem in the field of paper bleaching is the removal of lignin and its derivatives, which are linked to cellulose and xylan. Xylanases are more suitable in the paper and pulp industry than lignin-degrading systems. 相似文献
35.
Elik Aharonovsky Edward N. Trifonov 《Journal of biomolecular structure & dynamics》2013,31(3):237-242
Abstract Conserved protein sequence segments are commonly believed to correspond to functional sites in the protein sequence. A novel approach is proposed to profile the changing degree of conservation along the protein sequence, by evaluating the occurrence frequencies of all short oligopeptides of the given sequence in a large proteome database. Thus, a protein sequence conservation profile can be plotted for every protein. The profile indicates where along the sequences the potential functional (conserved) sites are located. The corresponding oligopeptides belonging to the sites are very frequent across many prokaryotic species. Analysis of a representative set of such profiles reveals a common feature of all examined proteins: they consist of sequence modules represented by the peaks of conservation. Typical size of the modules (peak-to-peak distance) is 25–30 amino acid residues. 相似文献
36.
目的 评价预防性应用微生态制剂(金双歧)在老年人抗生素相关性腹泻中的作用.方法 2010年5月至2013年2月应用抗生素的住院老年患者55例纳入观察对象,随机分为2组,预防组28例,给予微生态制剂双歧杆菌乳杆菌三联活菌片(金双歧)4片,3次/d,连续服用7~l0d;对照组27例,腹泻患者给予给予思密达3.76 g,3次/d.观察两组发生抗生素相关性腹泻情况,并进行肠道菌群分析.结果 预防组腹泻发生率、腹泻次数、腹泻持续时间与对照组比较,差异有统计学意义(P<0.05).两组腹泻病例的大便培养及常规检查无异常,大便涂片检查出现细菌计数减少或大便杆球菌比失调;预防组粪检结果杆球比例倒置、细菌计数减少、真菌感染例数等与对照组比较差异有统计学意义(P<0.05).结论 双歧杆菌乳杆菌三联活菌片(金双歧)可预防和减少老年人抗生素相关性腹泻的发生,减轻腹泻症状. 相似文献
37.
Dispersal moves individuals from patches where their immediate ancestors were successful to sites where their genotypes are untested. As a result, dispersal generally reduces fitness, a phenomenon known as “migration load.” The strength of migration load depends on the pattern of dispersal and can be dramatically lessened or reversed when individuals move preferentially toward patches conferring higher fitness. Evolutionary ecologists have long modeled nonrandom dispersal, focusing primarily on its effects on population density over space, the maintenance of genetic variation, and reproductive isolation. Here, we build upon previous work by calculating how the extent of local adaptation and the migration load are affected when individuals differ in their dispersal rate in a genotype‐dependent manner that alters their match to their environment. Examining a one‐locus, two‐patch model, we show that local adaptation occurs through a combination of natural selection and adaptive dispersal. For a substantial portion of parameter space, adaptive dispersal can be the predominant force generating local adaptation. Furthermore, genetic load may be largely averted with adaptive dispersal whenever individuals move before selective deaths occur. Thus, to understand the mechanisms driving local adaptation, biologists must account for the extent and nature of nonrandom, genotype‐dependent dispersal, and the potential for adaptation via spatial sorting of genotypes. 相似文献
38.
Erika L. Spaeth Christopher M. Booth Frank C. Marini 《Journal of visualized experiments : JoVE》2013,(79)
With the desire to understand the contributions of multiple cellular elements to the development of a complex tissue; such as the numerous cell types that participate in regenerating tissue, tumor formation, or vasculogenesis, we devised a multi-colored cellular transplant model of tumor development in which cell populations originate from different fluorescently colored reporter gene mice and are transplanted, engrafted or injected in and around a developing tumor. These colored cells are then recruited and incorporated into the tumor stroma. In order to quantitatively assess bone marrow derived tumor stromal cells, we transplanted GFP expressing transgenic whole bone marrow into lethally irradiated RFP expressing mice as approved by IACUC. 0ovarian tumors that were orthotopically injected into the transplanted mice were excised 6-8 weeks post engraftment and analyzed for bone marrow marker of origin (GFP) as well as antibody markers to detect tumor associated stroma using multispectral imaging techniques. We then adapted a methodology we call MIMicc- Multispectral Interrogation of Multiplexed cellular compositions, using multispectral unmixing of fluoroprobes to quantitatively assess which labeled cell came from which starting populations (based on original reporter gene labels), and as our ability to unmix 4, 5, 6 or more spectra per slide increases, we''ve added additional immunohistochemistry associated with cell lineages or differentiation to increase precision. Utilizing software to detect co-localized multiplexed-fluorescent signals, tumor stromal populations can be traced, enumerated and characterized based on marker staining.1 相似文献
39.
Ganive Bhinder Ho Pan Sham Justin M. Chan Vijay Morampudi Kevan Jacobson Bruce A. Vallance 《Journal of visualized experiments : JoVE》2013,(72)
This protocol outlines the steps required to produce a robust model of infectious disease and colitis, as well as the methods used to characterize Citrobacter rodentium infection in mice. C. rodentium is a gram negative, murine specific bacterial pathogen that is closely related to the clinically important human pathogens enteropathogenic E. coli and enterohemorrhagic E. coli. Upon infection with C. rodentium, immunocompetent mice suffer from modest and transient weight loss and diarrhea. Histologically, intestinal crypt elongation, immune cell infiltration, and goblet cell depletion are observed. Clearance of infection is achieved after 3 to 4 weeks. Measurement of intestinal epithelial barrier integrity, bacterial load, and histological damage at different time points after infection, allow the characterization of mouse strains susceptible to infection.The virulence mechanisms by which bacterial pathogens colonize the intestinal tract of their hosts, as well as specific host responses that defend against such infections are poorly understood. Therefore the C. rodentium model of enteric bacterial infection serves as a valuable tool to aid in our understanding of these processes. Enteric bacteria have also been linked to Inflammatory Bowel Diseases (IBDs). It has been hypothesized that the maladaptive chronic inflammatory responses seen in IBD patients develop in genetically susceptible individuals following abnormal exposure of the intestinal mucosal immune system to enteric bacteria. Therefore, the study of models of infectious colitis offers significant potential for defining potentially pathogenic host responses to enteric bacteria. C. rodentium induced colitis is one such rare model that allows for the analysis of host responses to enteric bacteria, furthering our understanding of potential mechanisms of IBD pathogenesis; essential in the development of novel preventative and therapeutic treatments. 相似文献
40.
Ubaldo E Martinez-Outschoorn Joseph M Curry Ying-Hui Ko Zhao Lin Madalina Tuluc David Cognetti Ruth C Birbe Edmund Pribitkin Alessandro Bombonati Richard G Pestell Anthony Howell Federica Sotgia Michael P Lisanti 《Cell cycle (Georgetown, Tex.)》2013,12(16):2580-2597
Here, we developed a model system to evaluate the metabolic effects of oncogene(s) on the host microenvironment. A matched set of “normal” and oncogenically transformed epithelial cell lines were co-cultured with human fibroblasts, to determine the “bystander” effects of oncogenes on stromal cells. ROS production and glucose uptake were measured by FACS analysis. In addition, expression of a panel of metabolic protein biomarkers (Caveolin-1, MCT1, and MCT4) was analyzed in parallel. Interestingly, oncogene activation in cancer cells was sufficient to induce the metabolic reprogramming of cancer-associated fibroblasts toward glycolysis, via oxidative stress. Evidence for “metabolic symbiosis” between oxidative cancer cells and glycolytic fibroblasts was provided by MCT1/4 immunostaining. As such, oncogenes drive the establishment of a stromal-epithelial “lactate-shuttle”, to fuel the anabolic growth of cancer cells. Similar results were obtained with two divergent oncogenes (RAS and NFκB), indicating that ROS production and inflammation metabolically converge on the tumor stroma, driving glycolysis and upregulation of MCT4. These findings make stromal MCT4 an attractive target for new drug discovery, as MCT4 is a shared endpoint for the metabolic effects of many oncogenic stimuli. Thus, diverse oncogenes stimulate a common metabolic response in the tumor stroma. Conversely, we also show that fibroblasts protect cancer cells against oncogenic stress and senescence by reducing ROS production in tumor cells. Ras-transformed cells were also able to metabolically reprogram normal adjacent epithelia, indicating that cancer cells can use either fibroblasts or epithelial cells as “partners” for metabolic symbiosis. The antioxidant N-acetyl-cysteine (NAC) selectively halted mitochondrial biogenesis in Ras-transformed cells, but not in normal epithelia. NAC also blocked stromal induction of MCT4, indicating that NAC effectively functions as an “MCT4 inhibitor”. Taken together, our data provide new strategies for achieving more effective anticancer therapy. We conclude that oncogenes enable cancer cells to behave as selfish “metabolic parasites”, like foreign organisms (bacteria, fungi, viruses). Thus, we should consider treating cancer like an infectious disease, with new classes of metabolically targeted “antibiotics” to selectively starve cancer cells. Our results provide new support for the “seed and soil” hypothesis, which was first proposed in 1889 by the English surgeon, Stephen Paget. 相似文献