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1.
Trichomoniasis is now an important health problem in developing countries. Although metronidazole has so far been widely used to treat this disease, the prevalence of metronidazole-resistant protozoa and unpleasant adverse effects have been found. In this study, natural products purified from Thai plants were, therefore, investigated for their effectiveness against Trichomonas vaginalis. The minimal inhibitory concentrations for all beta-glycosides against Trichomonas vaginalis at 24 h were in a range of 6.25-12.5 microM. In addition, torvoside A and H were found to be more potent than their corresponding aglycones, deglucosylated torvoside A and H, while other beta-glycosides were generally as active as their corresponding aglycones. The cytotoxicity of these compounds was also determined. Except for dalcochinin, none of the tested compounds showed cytotoxicity against Vero and cancer cell lines (KB and MCF-7), having IC(50) values greater than 50 microg/ml. In conclusion, beta-glycosides and several aglycones showed selective inhibition against Trichomonas vaginalis without harmful effect to mammalian cells.  相似文献   

2.
The PSO3 gene of Saccharomyces cerevisiae was molecularly cloned by complementing the cold-sensitivity phenotype of a pso3-1 mutant and was found to be allelic to RNR4, encoding one of the two DNA damage-inducible small subunits of the ribonucleotide reductase (RNR) complex. Compared to a rnr4Δ mutant that allows only very little mutation induction at very low doses of 254nm ultraviolet light (UVC), the pso3-1 mutant allele confers leakiness in that it permits some DNA damage-induced mutagenesis at low doses of UVC. Similarly, the pso3 mutant is slightly less sensitive to UVC than an rnr4Δ mutant. Cloning and sequencing of the RNR4 locus of the pso3-1 mutant revealed that its intermediate phenotype is attributable to a G → A transition at nucleotide 352, leading to replacement of glycine by arginine [G118R] in the mutant’s protein. Both RNR4 mutant alleles confer significantly less sensitivity to UVC than mutant alleles of non-UVC-mutable REV3, indicating that, apart from nucleotide excision repair, RAD6-dependent error-free DNA repair may still be functional. The phenotype of a strongly reduced UVC-induced mutagenesis for rnr4 mutant alleles has not yet been described; it suggests the importance of this gene for a fully functional RNR providing correct amounts of DNA precursor molecules, thereby, allowing translesion synthesis (error-prone) of UVC-damaged DNA. Stationary phase cells of the rnr4Δ mutant, but not of the original pso3-1 mutant, are swollen with a fourfold to eightfold increase in volume. The central role of RNR in DNA precursor metabolism and its complex regulation allow for several modes of suppression that may influence the phenotypes of RNR4 mutants, especially those containing the leaky pso3-1 mutant allele.  相似文献   

3.
This study analyzed the prevalence and antibiotic susceptibility of urogenital Ureaplasma urealyticum and Mycoplasma hominis isolated in Xi’an, China. A total of 2161 individuals from 2011 to 2015 were included, and antibiotic susceptibility tests were performed by using the Mycoplasma IST kit. Of the individuals studied, 1018 (47.11 %) were identified to be positive for urogenital mycoplasmas. The single U. urealyticum, single M. hominis, and dual U. urealyticum and M. hominis infections accounted for 772 (75.83 %), 66 (6.48 %), and 180 (17.68 %), respectively. The total positive rate was higher in females than in males (58.76 % vs. 28.86 %, p?<?0.001). The highest total positive rate (48.88 %) was observed in individuals aged 25 years to 30 years. In symptomatic and asymptomatic individuals, the positive rates were both higher in females than in males (67.36 % vs. 31.02 %, p?<?0.001 and 42.58 % vs. 7.69 %, p?<?0.001, respectively) and individuals aged 25 years to 30 years, and those aged 30 years to 35 years had the highest positive rates (54.35 and 57.14 %, respectively). The U. urealyticum and M. hominis identified from single or dual infections displayed low resistance rates to josamycin, doxycycline, and minocycline (<10 %) in both the symptomatic and asymptomatic groups. These results suggest that females and individuals with symptoms and younger age had higher mycoplasma infection rates and that josamycin, doxycycline, and minocycline may be recommended for the clinical treatment of patients infected with urogenital mycoplasmas, irrespective of the symptoms.  相似文献   

4.
Mosquitoes are important vectors of many infectious diseases. Bacillus sphaericus (Bs) is an ideal larvicide and has attracted more and more attention, recently. However, the fundamental research of its application is very limited, especially on the subsequent impact of Bs exposure on mosquito’s fecundity and resistance emergence. Through bioassay, LC50 and LC95 of Bs in killing Anopheles dirus larvae were determined as 9.793 ± 1.878 IU/L and 62.4 ± 6.438 IU/L at 48 h posttreatment, 7.128 ± 0.913 IU/L and 34.385 ± 12.547 IU/L at 72 h post treatment, respectively. After being treated with a sub-lethal dose of Bs, gravidity, oviposition, hatch, pupation, and eclosion of the surviving mosquitoes were counted and analyzed to elucidate the subsequent effects of Bs exposure on the reproductive capacity of A. dirus. The result interestingly showed that the exposure of Bs significantly reduced the oviposition ability of the surviving A. dirus, without effect on egg formation/gravidity, hatch, pupation, and eclosion. The surviving mosquitoes were also maintained routinely for generations to test the sustained effect of Bs exposure on the fecundity of the offsprings. After conventional breeding for generations, the capacity of egg laying totally recovered. To explore the rules of resistance development, bioassays were performed after treatment twice with a sub-lethal dose of Bs on two continuous generations of A. dirus larvae. The killing efficacies between the Bs treated group and control group were compared. The results showed that LC50 and LC95 increased by 4.35- and 7.37-folds after treatment with the sub-lethal dose of Bs on two consecutive generations, respectively. The results indicated that A. dirus was sensitive to Bs, which could reduce oviposition of the surviving A. dirus. The subsequent effect might help to further decrease the mosquito population. However, a sub-lethal dose of Bs exposure could easily cause resistance development. Our study provides a dose standard and reference for the rational use of Bs, which will be helpful for mosquito control.  相似文献   

5.
Integration of repair tissue is a key indicator of the long-term success of cell-based therapies for cartilage repair. The objective of this study was to compare the in vitro chondrogenic differentiation and integration of agarose hydrogels seeded with either chondrocytes or bone marrow-derived mesenchymal stem cells (MSCs) in defects created in cartilage explants. Chondrocytes and MSCs were isolated from porcine donors, suspended in 2% agarose and then injected into cylindrical defects within the explants. These constructs were maintained in a chemically defined medium supplemented with 10 ng/mL of TGF-β3. Cartilage integration was assessed by histology and mechanical push-out tests. After 6 weeks in culture, chondrocyte-seeded constructs demonstrated a higher integration strength (64.4 ± 8.3 kPa) compared to MSC-seeded constructs (22.7 ± 5.9 kPa). Glycosaminoglycan (GAG) (1.27 ± 0.3 vs. 0.19 ± 0.03 kPa) and collagen (0.31 ± 0.08 vs. 0.09 ± 0.01 kPa) accumulation in chondrocyte-seeded constructs was greater than that measured in the MSC-seeded group. The GAG, collagen, and DNA content of both chondrocyte- and MSC-seeded hydrogels cultured in cartilage explants was significantly lower than control constructs cultured in free swelling conditions. The results of this study suggest that the explant model may constitute a more rigorous in vitro test to assess MSC therapies for cartilage defect repair.  相似文献   

6.
7.
Human defensin-α1 is a biologically active peptide exhibiting a dose-dependent trypanocidal effect in vitro against trypomastigotes and amastigotes of Trypanosoma cruzi line Tulahuen. This effect is determined by fragmentation of parasite DNA reducing the capacity of passaged T. cruzi to invade HeLa cells.  相似文献   

8.
9.
In the present study, larvae of Ascaris suum and Trichuris muris were investigated by light and electron microscopy after incubation in a hatching medium containing 89% phosphate-buffered saline (pH 7.4), 10% RPMI-1640 and 1% sodiumhypochlorite at 40 and 37 degrees C, respectively. The larvae were obtained from fertilised eggs of the worms during defined phases of development (A. suum, 36th-50th day of development; T. muris, once a week from week 16 to 20). Light and electron micrographs of the larvae gave evidence that the third larval stage of A. suum is probably the infectious stage. The first moult of the larvae had already taken place before the 36th day of incubation starting at day 1. After 36 days of incubation, only the second larval stage was found within eggs. Some of these larvae were coated by a separated sheath so that a second moult of the larvae is reasonable. On the other hand, no sheathed larvae of T. muris were found in the eggs incubated for 20 weeks in distilled water. No signs of moult were seen for 20 weeks neither on light nor on the electron micrographs. Therefore, in T. muris, the first larval stage is the infectious stage, which was proven by means of re-infections of mice 16, 18 or 20 weeks after incubation of the eggs.  相似文献   

10.
While the counterselectable Schizosaccharomyces pombe ura4 + gene can be used to prepare a site in the S. pombe genome to receive an unmarked mutant allele (loss of ura4 + confers 5FOA-resistant (5FOAR) growth), the desired unmarked knock-in strains are generally outnumbered by spontaneously arising 5FOAR mutants. Relative to the same approach using the homologous URA3 + gene in Saccharomyces cerevisiae, knock-ins in S. pombe are harder to identify due to a lower efficiency of homologous recombination and a relatively high background of spontaneous 5FOAR colonies. To develop an improved method for identifying cells receiving unmarked mutant alleles, we first determined that 5FOAR strains carry mutations in either of two genes; ura4 + and ura5 +. We then cloned the S. pombe ura5 + orotate phosphoribosyltransferase gene and constructed a 2.1 kb cassette containing ura5 + together with the S. pombe lys7 + gene. Using this doubly marked cassette to disrupt the sck1 + kinase gene, we can distinguish between strains created by homologous knock-in of unmarked wild-type or kinase-dead alleles and spontaneously arising ura4 and ura5 mutants by screening 5FOAR colonies for the loss of the lys7 + marker. The utility of this system, especially when the phenotype for the strain carrying the knock-in allele is indistinguishable from that of the disruption strain, is borne out by the fact that ~95% of 5FOAR colonies in our studies arose from background ura4 and ura5 mutations.  相似文献   

11.
Yan Z  Hull CM  Sun S  Heitman J  Xu J 《Current genetics》2007,51(3):187-195
In the great majority of sexual eukaryotes, mitochondrial genomes are inherited almost exclusively from a single parent. While many hypotheses have been proposed to explain this phenomenon, very little is known about the genetic elements controlling uniparental mitochondria inheritance. In the bipolar, isogamous basidiomycete yeast Cryptococcus neoformans, progeny from crosses between strains of mating type a (MATa) and mating type α (MATα) typically inherit mitochondrial DNA (mtDNA) from the MATa parent. We recently demonstrated that a mating type α (MATα)-specific gene SXI1α, controls mitochondrial inheritance in C. neoformans. Here, we show that another homeodomain gene SXI2a in the alternative mating type MATa is also required for uniparental mtDNA inheritance in this fungus. Disruption of SXI2a resulted in biparental mtDNA inheritance in the zygote population with significant numbers of progeny inheriting mtDNA from the MATa parent, the MATα parent, and both the MATa and the MATα parents. In addition, progeny from same-sex mating between MATα strains showed a biparental mitochondrial inheritance pattern. Our results suggest that SXI1α and SXI2a coordinately control uniparental mitochondrial inheritance in C. neoformans.  相似文献   

12.
RNA primer removal from Okazaki fragments during lagging-strand replication and the excision of damaged DNA bases requires the action of structure-specific nucleases, such as the mammalian flap endonuclease 1 (FEN-1). This nuclease contains two conserved motifs enriched with acidic amino acid residues that are important for catalytic function. Similar motifs have been identified in nucleases found in viruses, archebacteria, eubacteria, and in eukaryotes ranging from yeast to humans. Unique among these proteins, the putative FEN-1 homologue in Escherichia coli is contained within the N-terminal region of the DNA polymerase I (PolN). To demonstrate that the cellular functions of FEN-1 reside in PolN, we cloned and expressed the amino terminal domain (323 amino acid residues) of PolI in a Saccharomyces cerevisiae strain lacking the FEN-1 homologue RAD27. Overexpression of PolN suppressed, to varying degrees, phenotypes associated with a rad27 null strain. These include temperature sensitivity, Okazaki fragment processing, a mutator phenotype, a G2/M cell cycle arrest, minichromosome loss, and methyl methane sulfonate sensitivity. We purified Rad27 and PolN proteins in order to determine whether differences in their intrinsic nuclease activities or interaction with proliferating cell nuclear antigen (PCNA) could explain the partial suppression of some phenotypes. We found that the in vitro nuclease activities of Rad27 were more potent than those of PolN and the activity of Rad27, but not PolN, was stimulated by PCNA. We conclude that the N-terminal nuclease domain of E. coli polymerase I encodes a functional homologue of FEN-1.  相似文献   

13.
This study considered the systems controlling sexual and self-recognition in Amylostereum areolatum, a homobasidiomycetous symbiont of the Sirex woodwasp. To investigate the structure and organization of these systems in A. areolatum, we identified a portion of a putative homologue (RAB1) of the pheromone receptor genes of Schizophyllum commune and Coprinus cinereus, and a portion of a putative homologue of the S. commune mitochondrial intermediate peptidase (mip) gene. Diagnostic DNA-based assays for mating-type were developed and their application confirmed that the fungus has a heterothallic tetrapolar mating system. Segregation analysis showed that RAB1 is linked to mating-type B, while mip is linked to mating-type A. The results of sexual and vegetative compatibility tests suggest that sexual recognition in A. areolatum is controlled by two multiallelic mat loci, while self-recognition is controlled by at least two multiallelic het loci. Therefore, despite the association of A. areolatum with the woodwasp and the unique mixture of sexual and clonal reproduction of the fungus, both recognition systems of the fungus appear to be similar in structure and function to those of other homobasidiomycetes. This is the first report regarding the genes controlling recognition of a homobasidiomycete involved in an obligate mutualistic relationship with an insect.  相似文献   

14.
The Acremonium chryrsogenum cefT gene encoding a membrane protein of the major facilitator superfamily implicated in the cephalosporin biosynthesis in A. chrysogenum was introduced into Penicillium chrysogenum Wisconsin 54-1255 (a benzylpenicillin producer), P. chrysogenum npe6 pyrG (-) (a derivative of Wisconsin 54-1255 lacking a functional penDE gene) and P. chrysogenum TA98 (a deacetylcephalosporin producer containing the cefD1, cefD2, cefEF and cefG genes from A. chrysogenum). RT-PCR analysis revealed that the cefT gene was expressed in P. chrysogenum strains. HPLC analysis of the culture broths of the TA98 transformants showed an increase in the secretion of deacetylcephalosporin C and hydrophilic penicillins (isopenicillin N and penicillin N). P. chrysogenum Wisconsin 54-1255 strain transformed with cefT showed increased secretion of the isopenicillin N intermediate and a drastic decrease in the benzylpenicillin production. Southern and northern blot analysis indicated that the untransformed P. chrysogenum strains contain an endogenous gene similar to cefT that may be involved in the well-known secretion of the isopenicillin N intermediate. In summary, the cefT transporter is a hydrophilic beta-lactam transporter that is involved in the secretion of hydrophilic beta-lactams containing alpha-aminoadipic acid side chain (isopenicillin N, penicillin N and deacetylcephalosporin C).  相似文献   

15.
16.
A newly cloned Helicoverpa armigera nucleopolyhedrovirus (HearNPV) from Kenya, HearNPV-NNg1, has a higher insecticidal activity than HearNPV-G4, which also exhibits lower insecticidal activity than HearNPV-C1. In the search for genes and/or nucleotide sequences that might be involved in the observed virulence differences among Helicoverpa spp. NPVs, the entire genome of NNg1 was sequenced and compared with previously sequenced genomes of G4, C1 and Helicoverpa zea single-nucleocapsid NPV (Hz). The NNg1 genome was 132,425 bp in length, with a total of 143 putative open reading frames (ORFs), and shared high levels of overall amino acid and nucleotide sequence identities with G4, C1 and Hz. Three NNg1 ORFs, ORF5, ORF100 and ORF124, which were shared with C1, were absent in G4 and Hz, while NNg1 and C1 were missing a homologue of G4/Hz ORF5. Another three ORFs, ORF60 (bro-b), ORF119 and ORF120, and one direct repeat sequence (dr) were unique to NNg1. Relative to the overall nucleotide sequence identity, lower sequence identities were observed between NNg1 hrs and the homologous hrs in the other three Helicoverpa spp. NPVs, despite containing the same number of hrs located at essentially the same positions on the genomes. Differences were also observed between NNg1 and each of the other three Helicoverpa spp. NPVs in the diversity of bro genes encoded on the genomes. These results indicate several putative genes and nucleotide sequences that may be responsible for the virulence differences observed among Helicoverpa spp., yet the specific genes and/or nucleotide sequences responsible have not been identified.  相似文献   

17.
A new species of sandfly is described from limestone caves in Thailand. The inclusion of this species in the subgenus Euphlebotomus is justified on the basis of characters of the male genitalia (paramere, basal lobe). The male–female gathering in the same taxon is based on ecological (cavernicolous species), morphological (length of male genital filaments and female spermathecal ducts) and molecular (homology of cytochrome b mt DNA sequences) criteria. A differential diagnosis between Phlebotomus mascomai n. sp. and P. argentipes Annandale & Brunetti, the vector of Leishmania donovani (Laveran & Mesnil) in India, is proposed based on several morphological characters like antennal formula and genitalia.  相似文献   

18.
Ex vivo production of IFN-γ, TNF-α, and IL-6 by mouse peritoneal macrophages was studied during successive infection with the vaccine strain M. bovis BCG and virulent strain M. tuberculosis H37Rv. The increase in the concentrations of TNF-α and IL-6 did not depend on the sequence of macrophage infection with the vaccine or virulent strain, but was related to the presence of the vaccine strain M. bovis BCG in the medium. IFN-γ production depended on infection of macrophages with the virulent strain M. tuberculosis H37Rv. The concentration of IFN-γ was maximum during primary infection with the virulent strain and did not increase after successive infection with the virulent and vaccine strain. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 144, No. 11, pp. 552–555, November, 2007  相似文献   

19.
Plant extracts, especially botanical insecticides, are currently studied more and more because of the possibility of their use in plant protection. Biological activity of five solvent plant extracts were studied using fourth instar larvae of gram pod borer Helicoverpa armigera (Lepidoptera: Noctuidae), cotton leaf roller Sylepta derogata (Lepidoptera: Pyralidae) and malaria vector Anopheles stephensi (Diptera: Culicidae). Antifeedant and larvicidal activity of acetone, chloroform, ethyl acetate, hexane and methanol peel, leaf and flower extracts of Citrus sinensis, Ocimum canum, Ocimum sanctum and Rhinacanthus nasutus were used in this study. During preliminary screening, the extracts were tested at 1,000 ppm concentration. The larval mortality was observed after 24 h of exposure. All extracts showed moderate larvicidal effects; however, the highest larval mortality was found in peel chloroform extract of C. sinensis, flower methanol extract of O. canum against the larvae of H. armigera (LC50 = 65.10,51.78, LC90 = 277.39 and 218.18 ppm), peel methanol extract of C. sinensis, flower ethyl acetate extract of O. canum and leaf acetone extract of O. sanctum against the larvae of S. derogata (LC50 = 20.27,58.21,36.66, LC90 =113.15,285.70 and 668.02 ppm), peel methanol extract of C. sinensis, leaf and flower ethyl acetate extracts of O. canum against the larvae of A. stephensi (LC50 = 95.74,101.53,28.96, LC90 = 303.20,492.43 and 168.05 ppm), respectively. These results suggest that the chloroform and methanol extract of C. sinensis, ethyl acetate flower extracts of O. canum and acetone extract of O. sanctum have the potential to be used as an ideal eco-friendly approach for the control of the agricultural pests H. armigera, S. derogata and medically important vector A. stephensi.  相似文献   

20.
Acute lung injury (ALI) is characterized by widespread inflammation in the lungs and alveolar-capillary destruction, causing high morbidity and mortality. Cavidine, isolated from Corydalis impatiens, have been exhibited to have potent anti-inflammatory effects in previous studies. The purpose of this study was to evaluate the protective effect of cavidine on lipopolysaccharide (LPS)-induced ALI and to enunciate the underlying in vivo and in vitro mechanisms. Mice were intraperitoneally administrated with cavidine (1, 3, or 10 mg/kg) at 1 and 12 h, prior to the induction of ALI by intranasal administration of LPS (30 mg/kg). Blood samples, lung tissues, and bronchoalveolar lavage fluid (BALF) were harvested after LPS challenge. Furthermore, we used LPS-induced lung epithelial cells A549 to examine the mechanism of cavidine to lung injury. The results showed that pretreatment with cavidine significantly decreased lung wet-to-dry weight (W/D) ratio, reduced pro-inflammatory cytokine levels including TNF-α and IL-6 in BALF and serum from LPS-stimulated mice, and attenuated lung histopathological changes. In addition, western blot results showed that cavidine inhibited the phosphorylation of nuclear factor-kappaB (NF-κB) p65 and IκBα induced by LPS. In conclusion, our results demonstrate that cavidine protects against LPS-induced acute lung injury in mice via inhibiting of pro-inflammatory cytokine TNF-α and IL-6 production and NF-κB signaling pathway activation. Taken together, cavidine may be useful for the prevention and treatment of pulmonary inflammatory diseases, such as ALI.  相似文献   

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