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981.
This research examines possible factors limiting pathogen development and reproduction in a novel host insect. The nematode Heterorhabditis marelatus and its symbiotic bacterium, Photorhabdus luminescens, kill 98% of nematode-treated Colorado potato beetle (CPB) prepupae, but the nematode reproduces in only 1-6% of beetles. We examined nematode/bacterial inhibition at each step of the normal developmental pathway to determine host feature(s) limiting nematode reproduction. We found that in vivo encapsulation of nematodes occurred in only 1.6% of CPB, and in 5% of in vitro hanging drops of hemolymph. Thus, the cellular defense system did not strongly limit nematode reproduction in the CPB. The symbiotic bacterium was negatively affected by a heat-labile factor found in the CPB's hemolymph which often caused the bacterium to switch from the primary form that produces antibiotics and nutrients necessary for the nematodes' development, to a secondary form that provides only limited nutrients. A 58 kDa protein was isolated and bioassayed for activity against P. luminescens, but caused a delay in bacterial growth rather than the primary-secondary form switch. Thus, the identity of the heat-labile factor could not be confirmed as being the 58 kDa protein. The heat-labile factor did not directly affect the nematode. The addition of lipids in the form of olive oil to heated CPB hemolymph allowed nematodes to reproduce in 17% of hanging drops, in contrast to zero reproduction in hemolymph without oil. Reproductive nematodes were smaller when grown in CPB hemolymph than in hemolymph of the highly susceptible Galleria mellonella. These data suggest that both the toxic heat-labile factor and a lack of appropriate nutrients alter the CPB-bacterium-nematode interaction. These factors preclude the use of this otherwise highly effective nematode-bacterial complex in the longterm control of the CPB. 相似文献
982.
Byrd CM Bolken TC Mjalli AM Arimilli MN Andrews RC Rothlein R Andrea T Rao M Owens KL Hruby DE 《Journal of virology》2004,78(22):12147-12156
By using a homology-based bioinformatics approach, a structural model of the vaccinia virus (VV) I7L proteinase was developed. A unique chemical library of approximately 51,000 compounds was computationally queried to identify potential active site inhibitors. The resulting biased subset of compounds was assayed for both toxicity and the ability to inhibit the growth of VV in tissue culture cells. A family of chemotypically related compounds was found which exhibits selective activity against orthopoxviruses, inhibiting VV with 50% inhibitory concentrations of 3 to 12 microM. These compounds exhibited no significant cytotoxicity in the four cell lines tested and did not inhibit the growth of other organisms such as Saccharomyces cerevisiae, Pseudomonas aeruginosa, adenovirus, or encephalomyocarditis virus. Phenotypic analyses of virus-infected cells were conducted in the presence of active compounds to verify that the correct biochemical step (I7L-mediated core protein processing) was being inhibited. Electron microscopy of compound-treated VV-infected cells indicated a block in morphogenesis. Compound-resistant viruses were generated and resistance was mapped to the I7L open reading frame. Transient expression with the mutant I7L gene rescued the ability of wild-type virus to replicate in the presence of compound, indicating that this is the only gene necessary for resistance. This novel class of inhibitors has potential for development as an efficient antiviral drug against pathogenic orthopoxviruses, including smallpox. 相似文献
983.
Long-lived poxvirus immunity, robust CD4 help, and better persistence of CD4 than CD8 T cells
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The currently used smallpox vaccine is associated with a high incidence of adverse events, and there is a serious need for a safe and effective alternative vaccine. Here, we carried out a longitudinal evaluation of vaccinia virus-specific CD4 and CD8 T cells in smallpox-vaccinated individuals by using a highly sensitive intracellular cytokine staining assay. Our results demonstrate that, in addition to the CD8 response, the smallpox vaccinations raised a robust CD4 response with a Th1-dominant cytokine profile. These CD4 T cells were stable and exhibited only a twofold contraction between peak effector and memory phases compared with an approximate sevenfold contraction for CD8 cells. A significant proportion of vaccinated individuals lost detectable CD8 memory while maintaining CD4 memory. After a booster immunization, these individuals generated a robust CD8 response, which some of them rapidly lost. Thus, the current smallpox vaccine provides long-lasting CD4 help that may be critical for long-lived B-cell memory. We suggest that the provision of adequate CD4 help for CD8 and humoral effector functions will be critical to the success of the next generation of smallpox vaccines. 相似文献
984.
Copaifera langsdorffii oleo-resin (CLOR) is a reputed herbal medicine used to combat gastrointestinal functional disorders. Our previous studies show that CLOR prevents gastric ulceration and promotes wound healing. This study examined the effects of CLOR on intestinal damage associated with mesenteric ischemia/reperfusion in rat. Wistar albino rats were divided into four groups of six in each. Group 1: Sham operated, Group 2: Vehicle + 45 min of ischemia followed by 60 min reperfusion (I/R), Groups 3 and 4: I/R + CLOR (200 and 400 mg /kg, p.o., respectively). All treatments were given 24 h, 12 h and 2 h before I/R. Animals were sacrificed at the end of reperfusion period and ileal tissue samples were obtained for biochemical analysis. Myeloperoxidase (MPO), an index of polymorphonuclear leukocytes; malondialdehyde (MDA), an end product of lipoperoxidation; catalase (CAT), an antioxidant enzyme; reduced glutathione (GSH), a key antioxidant; and nitrite, a marker of nitric oxide (NO) production were determined in ileum homogenates. The results show that I/R produces a significant increase in MDA content, MPO, and CAT activities with a significant decrease in GSH and an elevation in nitrite production, as compared to sham control. CLOR treatment caused significant attenuations in I/R-associated increases of MPO, MDA and CAT activities and on nitrite level. Besides, CLOR could effectively prevent the I/R-associated depletion of GSH. The data indicate that the oleo-resin has a protective action against I/R-induced intestinal tissue damage, which appeared to be, at least in part, due to an antioxidant and anti-lipid peroxidation mechanism. 相似文献
985.
The roles played by the non-covalent interactions have been investigated for a set of six TNF proteins and nine Interleukins. The stabilizing residues have been identified by a consensus approach using the concepts of available surface area, medium and long-range interactions and conservation of amino acid residues. The cation-pi interactions have been computed based on a geometric approach such as distance and energy criteria. We identified an average of 1 energetically significant cation-pi interactions in every 94 residues in TNF proteins and 1 in every 62 residues in Interleukins. In TNF proteins, the cationic groups Lys preferred to be in helix while Arg preferred to be in strand regions while in Interleukins the Arg residues preferred to be in helix and Lys preferred to be in strand regions. From the available surface area calculations, we found that, almost all the cation and pi residues in TNF proteins and Interleukins were either in buried or partially buried regions and none of them in the exposed regions. Medium and long-range interactions were predominant in both TNF proteins and Interleukins. It was observed that the percentage of stabilizing centers were more in TNF proteins as compared to the Interleukins, while the percentage of conserved residues were more in Interleukins than in TNF proteins. In the stabilizing residues Lys was observed to be a stabilizing residue in both TNF proteins and Interleukins. Among the aromatic group, Phe was seen to be a stabilizing residue in both TNF and Interleukins. We suggest that this study on the computation of cation-pi interactions in TNF proteins and Interleukins would be very helpful in further understanding the structure, stability and functional similarity of these proteins. 相似文献
986.
987.
The cDNA encoding N-terminal three immunoglobin-like domains of human M-CSFR was linked to His-tag and endoplasmic reticulum retention sequence (KDEL) before being inserted into the genome of tobacco plant, Nicotiana tabacum cv. NC-89, by Agrobacterium tumefaciens-mediated transformation. The insertion and expression of target gene were confirmed by PCR, ELISA, and Western blot. The recombinant M-CSFsR reached a maximum expression level of 1.92% of total soluble protein in transgenic tobacco plant leaf tissues. The recombinant M-CSFsR could be purified through a one-step IMAC process and its bioactivity was confirmed by the inhibition of colony formation of J6-1 cells. The results suggested that we successfully expressed a high level of bioactive human M-CSFsR in tobacco plants. 相似文献
988.
We have developed a software program that weights and integrates specific properties on the genes in a pathogen so that they may be ranked as drug targets. We applied this software to produce three prioritized drug target lists for Mycobacterium tuberculosis, the causative agent of tuberculosis, a disease for which a new drug is desperately needed. Each list is based on an individual criterion. The first list prioritizes metabolic drug targets by the uniqueness of their roles in the M. tuberculosis metabolome ("metabolic chokepoints") and their similarity to known "druggable" protein classes (i.e., classes whose activity has previously been shown to be modulated by binding a small molecule). The second list prioritizes targets that would specifically impair M. tuberculosis, by weighting heavily those that are closely conserved within the Actinobacteria class but lack close homology to the host and gut flora. M. tuberculosis can survive asymptomatically in its host for many years by adapting to a dormant state referred to as "persistence." The final list aims to prioritize potential targets involved in maintaining persistence in M. tuberculosis. The rankings of current, candidate, and proposed drug targets are highlighted with respect to these lists. Some features were found to be more accurate than others in prioritizing studied targets. It can also be shown that targets can be prioritized by using evolutionary programming to optimize the weights of each desired property. We demonstrate this approach in prioritizing persistence targets. 相似文献
989.
Das B Ramu R Venkateswarlu K Rao YK Reddy MR Ramakrishna KV Harakishore K Murty US 《化学与生物多样性》2006,3(2):175-179
Two new ent-clerodane-type diterpenoids, compounds 1 and 2, were isolated from the aerial parts of Pulicaria wightiana, together with three known constituents. Their structures were established based on spectroscopic data, and their antibacterial activities were evaluated (Table 2). 相似文献
990.
Screening for High-Temperature Tolerant Cotton Cultivars by Testing In Vitro Pollen Germination, Pollen Tube Growth and Boll Retention 总被引:5,自引:0,他引:5
Zhi Liu You-Lu Yuan Shao-Qing Liu Xiao-Nan Yu Li-Qun Rao 《植物学报(英文版)》2006,48(6):706-714
With radical global climate change and global warming, high temperature stress has become one of major factors exerting a major influence on crop production. In the cotton (Gossypium hirsutum L.)-growing areas of China, especially in the Yangtze River valley, unexpected periodic episodes of extreme heat stress usually occur in July and August, the peak time of cotton flowering and boll loading, resulting in lower boll set and lint yield. Breeding programs for screening high temperature-tolerant cotton germplasm and cultivars are urgent in order to stabilize yield in the current and future warmer weather conditions. In the present study, 14 cotton cultivars were quantified for in vitro pollen germination and pollen tube growth in response to temperatures ranging from 10 to 50 ℃ at 5 ℃ intervals. Different cotton genotypes varied in their in vitro pollen germination and pollen tube length responses to the different temperatures. Maximum pollen germination and pollen tube length ranged from 25.2% to 56.2% and from 414 to 682 μm, respectively.The average cardinal temperatures (Tmin, Topt, and Tmax) also varied among the 14 cultivars and were 11.8,27.3, and 42.7 ℃ for pollen germination and 11.8, 27.8, and 44.1 ℃ for maximum pollen tube length. Variations in boll retention and boll numbers per plant in field experiments were found for the 14 cotton cultivars and the boll retention and boll retained per plant on 20 August varied considerably in different years according to weather conditions. Boll retention on 20 August was highly correlated with maximum pollen germination (R2=0.84) and pollen tube length (R2=0.64). A screening method based on principle component analysis of the combination of pollen characteristics in an in vitro experiment and boll retention testing in the field environment was used in the present study and, as a result, the 14 cotton cultivars could be classified as tolerant, moderately tolerant, moderately susceptible and susceptible to high temperature. 相似文献