An unusual murine behavior following infection with log-phase Nocardia asteroides type 6 strain GUH-2 (Nocardia cyriacigeorgica GUH-2)

Nocardia asteroides (Nocardia cyriacigeorgica) strain GUH-2 infects the brains of mice following intravenous injection. Non-lethal infections resulted in a transitory increase of bacterial numbers in the brain followed by the development of permanent impaired movements at a time when bacteria appeared to be eliminated from the brain. These signs included headshake, rigidity, stooped posture, dyskinesia, retropulsion, and abnormal tail positioning in approximately 20% of infected animals. The attached video presents a typical mouse following infection with this organism, as compared to an age- and gender-matched uninfected control mouse.     

Nocardia asteroides culture filtrates cause dopamine depletion and cytotoxicity in PC12 cells

Experimental infection of BALB/c mice with the gram-positive bacterium Nocardia asteroides produces marked loss of nigrostriatal dopamine neurons, resulting in striatal dopamine depletion. To investigate the mechanism(s) responsible for this neuronal loss, we evaluated the influence of N. asteroides cell-free culture filtrates on rat pheochromocytoma PC12 cells, an in vitro model for dopamine neurons. Changes in cell viability and cell numbers were minimal after 24 h, but increased with longer incubation. In contrast, dopamine depletion occurred after 30 min incubation, and was greater with GUH-2 filtrate than with filtrate from the less virulent strain 10905. Incubation with the culture filtrate decreased viability in neuroblastoma and glioma cell lines, indicating that cytotoxic effects were not limited to dopaminergic cells. These findings suggest that the loss of nigrostriatal dopamine neurons and concomitant striatal dopamine depletion in Nocardia-infected mice may be due, at least in part, to the neurotoxicity of nocardial secretory products.     

Primary subcutaneous Nocardia asteroides infection in a renal allograft recipient

Nocardia asteroides is an important opportunistic pathogen in immunocompromised hosts. The primary infection is usually in the lungs and is followed by dissemination to other parts of the body. Primary subcutaneous infection with Nocardia asteroides has been reported rarely (three reports) and no such case has been reported in a renal transplant recipient. We describe here a case of renal transplant recipient who developed primary subcutaneous infection with Nocardia asteroides within one and half years of the transplantation. This revised version was published online in June 2006 with corrections to the Cover Date.     

Effect of temperature on the viability of Bacillus stearothermophilus

One of the obligate thermophilic bacteria, Bacillus stearothermophilus, was unable to grow at temperatures below 35° C. About 80% of the population in the bacterial culture died at the temperatures, and the same extent of loss in either of the activities of oxygen consumption or synthesis of protein or nucleic acid of the organisms was observed. With the progress of death of the organisms, reduced nicotinamide-adenine dinucleotide came to be oxidized by the organisms, enzymes such as fructose-1,6-diphosphate aldolase, when the organisms were washed with phosphate buffer, were leaked out of the organisms, and an increasing amount of ribonucleoprotein was released into the culture medium. The change of the membrane state was then suggested to be one of the possible causes for the death of the organisms at the temperatures.     

Enzymatic characterization of Nocardia spp. and related bacteria by API ZYM profile

Characterization of 62 isolates belonging to the genus Nocardia and related bacteria was obtained by using the API ZYM system. The difference in enzymatic profile should allow a relatively efficient, low-cost means to identify aerobic actinomycetes of clinical significance.     

Colonization and degradation of rubber pieces by Nocardia sp.

The growth of a Nocardia sp. occurs essentially on the insoluble rubber substrate and the cells are tightly bound to the rubber in the initial stage of the growth in spite of vigorous stirring of the cultures. The colonization of rubber pieces was followed by staining with Schiff reagent, and it was revealed that not only the thickness of rubber pieces, but also their length and width greatly influenced microbial colonization and degradation of natural rubber products. Among rubber pieces of various shapes, long strips were most rapidly covered by many microbial colonies and experienced the highest rate of rubber degradation. The rate of degradation (expressed by % weight loss) of the long strips of rubber was a linear function of surface area per unit weight of rubber. Thin and wide films of rubber were also rapidly colonized and degraded, while the colonization and degradation of short and narrow pieces were substantially slower and less extensive.     

Nocardia rhamnosiphila sp. nov., isolated from soil

In this study two actinomycete strains were isolated in Cape Town (South Africa), one from a compost heap (strain 202GMO^T) and the other from within the fynbos-rich area surrounded by the horseracing track at Kenilworth Racecourse (strain C2). Based on 16S rRNA gene sequence BLAST analysis, the strains were identified as members of the genus Nocardia. Phylogenetic analysis showed that the strains clustered together and are most closely related to Nocardia flavorosea NRRL B-16176^T, Nocardia testacea JCM 12235^T, Nocardia sienata IFM 10088^T and Nocardia carnea DSM 43397^T. This association was also supported by gyrB based phylogenetic analysis. The results of DNA–DNA hybridization and physiological tests allowed genotypic and phenotypic differentiation of both strains 202GMO^T and C2 from related species. However, their high DNA relatedness showed that they belong to the same species. Strain 202GMO^T was selected as the type strain to represent this novel species, for which the name Nocardia rhamnosiphila is proposed (=DSM 45147^T = NRRL B-24637^T).     

Decomposition of 14C-labelled lignin and phenols by a Nocardia sp.

A Gram-positive bacterium which was isolated from a Finnish soil and identified as a Nocardia sp., was able to decompose lignin and to assimilate lignin degradation products as a carbon source. It could release ¹⁴CO₂ from ¹⁴C-labelled methoxyl groups, side chains or ring carbons of coniferyl alcohol dehydropolymers (DHP) and from specifically ¹⁴C-labelled lignin of plant material. Furthermore, it could release ¹⁴CO₂ from phenolcarboxylic and cinnamic acids and alcohols labelled in the OCH₃, COOH groups, side chain or aromatic ring carbons.Non-Common Abbreviations Used DHP dehydropolymers of coniferyl alcohol     

A monoclonal antibody specific for the red-absorbing form of phytochrome

Characterisation of a new monoclonal antibody (mAb), designated LAS 41, directed against 124-kilodalton (kDa) etiolated-oat (Avena sativa L.) phytochrome, indicates that it recognises an epitope unique to the red-light-absorbing form, Pr. In a solid-phase enzyme-linked immunosorbent assay (ELISA), LAS 41 exhibits a seven- to eight-fold higher affinity for Pr than for the far-red-light-absorbing form of phytochrome, Pfr. In addition, in immunoprecipitation assays LAS 41 effectively precipitates 100% of phytochrome presented as Pr but only precipitates a maximum of 24.5% of phytochrome presented as Pfr. These values are indicative of binding exclusively to Pr. Peptide-mapping studies show that LAS 41 recognises and epitope located within a region 6–10 kDa from the aminoterminus of the phytochrome molecule. Since binding of LAS 41 to Pr induces alterations in the spectral properties of Pr, this indicates that at least part of the 4 kDa domain to which the antibody binds is essential for protein-chromophore interaction. Subsequent photoconversion of LAS 41-Pr complexes produces native Pfr spectra, with concomitant production of free antibody and antigen, as shown by a modified ELISA. The specificity of LAS 41 for Pr has facilitated the purification of Pfr which is free of contaminating Pr. This has enabled direct determination of the mole fraction of Pfr established by red light to be 0.874.Abbreviations ELISA enzyme-linked immunsorbent assay - kDa kilodalton - mAb monoclonal antibody - Pfr far-red-absorbing form of phytochrome - Pr red-absorbing form of phytochrome - SDS-PAGE sodium dodecyl sulphate polyacrylamide gel electrophoresis - (A) difference in absorbance (A ₆₆₅ ^Pr –A ₇₃₀ ^Pr )-(A ₆₆₅ ^Pfr –A ₇₃₀ ^Pfr ) - Ar/Afr spectral change ratio (SCR) - max mole fraction of Pfr following saturating red light     

Acid and alkali as aids in differentiation of pathogenic Nocardia and for their isolation from clinical specimens

The tolerance of Nocardia brasiliensis, Nocardia asteroides and Nocardia caviae to different concentrations of acids and alkalis was studied in this work. The purpose was to determine their use: (a) in the differentiation of species; (b) for decontamination of clinical materials during the isolation of pathogenic Nocardia. The results showed (1) that both 2 M NH₃ and 0.05 M H₂SO₄ solutions are useful for differentiating species. They can also be used as a complementary method for identification. (2) That both 1 M NH₃ and 0.05 M H₂SO₄ solutions were useful for decontamination of clinical materials during the isolation of pathogenic Nocardia.     

A note on the hatching and viability ofCeriodaphnia ephippia collected from lake sediment

Ephippia ofCeriodaphnia pulchella Sars were collected at 2 sites from successive sediment layers. Hatching observed in the laboratory gave information about the duration of their viability. Conclusions about the hatching situation in the lake were drawn from the ratio of intact to total ephippia at various lake depths. The results are discussed.     

基于Vortex模型分析金沙江水电开发对圆口铜鱼种群生存力的影响

圆口铜鱼(Coreius guichenoti)是长江上游特有鱼类,研究金沙江梯级水电开发背景下圆口铜鱼种群生存力变化对该物种的保护具有重要意义。Vortex模型在传统种群评估方法的基础上,引入种群年龄结构、繁殖体制、性比等因素,并考虑了环境、灾害、遗传等随机性对种群的影响,可模拟种群数量和生存力变化特征,是一种应用广泛的种群生存力分析模型。基于1981-2021年金沙江圆口铜鱼资源调查数据,采用Vortex模型构建了金沙江中下游干流大规模水电开发前、后两个时期圆口铜鱼种群生存力分析模型,考虑了水电开发下,水温、流速等环境变化对圆口铜鱼种群的影响,对比了两个时期圆口铜鱼种群数量、结构及生存力等。结果表明:水电开发前,严重过度捕捞对圆口铜鱼种群产生了显著威胁,10年内种群数量下降了12.5%,百年内种群内禀增长率为0.085,种群的生存和繁殖能力较稳定,灭绝概率为1%;水电开发后,10年内种群数量下降了42%,百年内种群内禀增长率降至-0.087,灭绝概率增大至36.1%,种群平均灭绝时间为83.7年,圆口铜鱼抵抗环境干扰的种群恢复能力降低,种群生存力受到影响。     

Quantification of Saccharomyces cerevisiae viability using BacLight

Yeast viability can be accurately quantified using BacLight, a kit which so far has been used only for bacterial analysis. Upon staining, viable cells can be differentiated from non-viable ones by either confocal laser scanning microscopy (CLSM), epifluorescence microscopy, or flow cytometry. Using Saccharomyces cerevisiae as a model, viabilities quantified by CLSM deviated an average of 1.7% from the actual data, and those determined by flow-cytometry by 1.4%.     

PD-1对伯氏疟原虫感染小鼠体液免疫应答的影响

利用伯氏疟原虫Plasmodium berghei ANKA(P.b ANKA)感染BALB/c小鼠,PD-1单抗阻断后,流式细胞术检测脾脏浆细胞、滤泡辅助性T细胞(Tfh)数量。qRT-PCR检测IL-21、IL-10和IL-6 mRNA水平,ELISA检测血清抗体,以探讨程序性死亡受体-1(programmed cell death-1, PD-1)在疟原虫初次感染中对体液免疫应答的影响。结果发现,PD-1单抗阻断加速了P.b ANKA感染小鼠的死亡。与对照组相比,PD-1阻断组感染后第12天短寿浆细胞(CD138~+CD44~+)数量明显降低(P0.05),长寿浆细胞(CD138~+CD44~-、CD138~-CD44~+)和Tfh(CD4~+CXCR5~+)细胞数量无差异性改变,脾细胞IL-21的mRNA水平明显下降(P0.05),血清抗裂殖子表面蛋白(merozoite surface protein, MSP)-1特异性IgG无明显改变。P.b ANKA感染中PD-1通路可能通过影响Tfh分泌IL-21进而干扰浆细胞数量影响体液免疫应答。     

Purification,characterization and regulation of a monomeric l-phenylalanine dehydrogenase from the facultative methylotroph Nocardia sp. 239

In Nocardia sp. 239 d-phenylalanine is converted into l-phenylalanine by an inducible amino acid racemase. The further catabolism of this amino acid involves an NAD-dependent l-phenylalanine dehydrogenase. This enzyme was detected only in cells grown on l- or d-phenylalanine and in batch cultures highest activities were obtained at relatively low amino acid concentrations in the medium. The presence of additional carbon- or nitrogen sources invariably resulted in decreased enzyme levels. From experiments with phenylalanine-limited continuous cultures it appeared that the rate of synthesis of the enzyme increased with increasing growth rates. The regulation of phenylalanine dehydrogenase synthesis was studied in more detail during growth of the organism on mixtures of methanol and l-phenylalanine. Highest rates of l-phenylalanine dehydrogenase production were observed with increasing ratios of l-phenylalanine/methanol in the feed of chemostat cultures. Characteristic properties of the enzyme were investigated following its (partial) purification from l- and d-phenylalanine-grown cells. This resulted in the isolation of enzymes with identical properties. The native enzyme had a molecular weight of 42 000 and consisted of a single subunit; it showed activity with l-phenylalanine, phenylpyruvate, 4-hydroxyphenyl-pyruvate, indole-3-pyruvate and -ketoisocaproate, but not with imidazolepyruvate, d-phenylalanine and other l-amino acids tested. Maximum activities with phenylpyruvate (310 mol min^-1 mg^-1 of purified protein) were observed at pH 10 and 53°C. Sorbitol and glycerol stabilized the enzyme.Abbreviations RuMP ribulose monophosphate - HPS hexulose-6-phosphate synthase - HPT hexulose-6-phosphate isomerase - FPLC fast protein liquid chromatography     

The effect of desiccation on viability and phospholipid composition of Acer sacharinum L. seeds

Summary Phospholipid biosynthesis and composition in desiccation-intolerant silver maple seeds were determined with respect to the seed's germinability and electrolyte leakage during the desiccation process. An evident increase in the biosynthesis of phospholipids occurs during the period of high viability of the seeds. The electrolyte leakage level during this period is rather low. The more the seed's viability declines the lower is the biosynthesis of phospholipids and the higher the permeability of membranes to solutes. The total content of phospholipids as well as the content of particular phospholipids decreases with a simultaneous increase of phosphatidic acid. Presented data support the hypothesis that membrane destruction takes place during loss of water in desiccation-intolerant seeds. The nature of cell membrane damage is discussed.     

Genetic differentiation in the viability of sibling species of Rhagoletis fruit flies on host plants, and the influence of reduced hybrid viability on reproductive isolation

Prior theoretical studies have analyzed host-associated factors restricting gene flow between phytophagous insect species, and simulated evolutionary changes in components of fitness contributing to the adaptation of species to distinct host plants. However, there is relatively little empirical information on these topics. In this paper we report data on two host-associated sibling species of tephritid fruit flies, Rhagoletis mendax (Curran) and R. pomonella (Walsh) which infest ericaceous and rosaceous host plants, respectively. To test the hypothesis that these species have evolved viability differences on alternate hosts which can contribute to the restriction of gene flow between them, we measured the larval-to-adult viability of R. mendax, R. pomonella and F₁ interspecific hybrid progeny reared on naturally-growing, highbush blueberry (Vaccinium corymbosum L.) and apple (Malus pumila Miller = Pyrus malus L.) plants in the field. Our results indicate that genetic changes associated with the adaptation of these species to distinct host plants could also cause reduced fitness of interspecific hybrids, and thereby restrict interspecific gene flow. Fewer interspecific hybrids survived to adulthood than either R. pomonella progeny reared in apples or R. mendax progeny reared in blueberries. These differences in the viability of progeny from hybrid versus conspecific crosses can substantially restrict gene flow between R. mendax and R. pomonella flies, and may be an important factor influencing their reproductive isolation. Genetic differences among hybrid and conspecific crosses were also detected for the lengths, widths and weights and weights of pupae reared from blueberries and apples. In a second experiment, the ovipositional preferences of R. mendax and R. pomonella flies were recorded on host plants inside a field cage. R. mendax flies had a strong ovipositional preference for blueberries over apples, whereas R. pomonella flies readily oviposited in both host fruits. Other studies have determined that R. pomonella flies also oviposit in the fruits of several other plants which are not hosts under field conditions; this behavior can promote host shifts to new plants by flies with suitable plant-finding and viability traits. Our results support the theoretical assumption that divergence in host-acceptance behaviors and viabilities on host plants are key aspects of evolutionary differentiation among closely-related taxa of phytophagous parasitic insects. We discuss our results in the context of other traits that can restrict gene flow between R. mendax and R. pomonella, and in relation to the findings of other studies on the evolution of host plant use.

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Résumé Bien que des études théoriques aient analysé tant l'influence des éléments de l'adaptabilité sur l'évolution de l'utilisation d'une plante par un insecte phytophage que les caractéristiques de l'hôte limitant le flux génique entre espèces voisines, peu de données concrètes concernent ces aspects.Nos résultats portent sur deux espèces jumelles de téphritides, R. mendax Curran et R. pomonella Walsh qui vivent respectivement sur éricacées et rosacées. Pour vérifier si des différences dans le comportement d'acceptation des hôtes et dans leur survie sur des hôtes alternatifs sont apparues chez ces espèces jumelles, nous avons observé dans la nature le comportement de ponte et la survie des 2 espèces et des F₁ hybrides interspécifiques sur Vaccinium corymbosum L. et Malus pumila Miller = Pyrus mali L. Le pourcentage de pondeuses, le nombre d'ufs pondus par femelle et la survie des larves ont été plus élevés pour R. mendax que pour R. pomonella sur V. corymbosum, tandis que le contraire était observé sur M. pumila. Des différences génétiques ont été observées pour la mortalité avant la nymphose et avant la mue imaginale lors des croisements hybrides et interspécifiques. Les descendants mâles et femelles de R. mendax ont mieux survécu sur V. corymbosum, tandis que la survie de la descendance des croisements entre R. pomonella était la même sur les deux plantes. Un peu plus d'hybrides ont atteint le stade adulte sur pomme que de descendants de croisements intraspécifiques, et la mortalité des descendants de femelles de R. pomonella sur V. corymbosum était plus élevée que celle des femelles de R. mendax. Les différences de survie et de comportement de ponte de R. mendax et de R. pomonella peuvent limiter le flux génique entre les 2 espèces. Des différences génétiques peuvent être observées quant à la taille et au poids des pupes de R. mendax et R. pomonella. Ces résultats confortent l'hypothèse que des divergences évolutives du comportement d'acceptation de l'hôte et de mortalité sur hôtes respectifs sont des facteurs clés séparant des taxa très voisins d'insectes parasites phytophages. La discussion intègre les autres aspects pouvant limiter le flux génique entre R. mendax et R. pomonella et les autres hypothèses sur l'évolution de l'exploitation des plantes hôtes.

     

Orchestration of the protective immune response to intracellular bacteria: Francisella tularensis as a model organism

Abstract Francisella tularensis is used as a model organism in studies of mechanisms behind the induction of a protective T-cell response in the mammalian host. Protective immunity is associated with a CD4 and CD8 T-cell response towards a mosaic of proteins of F. tularensis and due to HLA restriction, each individual selects her own mosaic. No single protein has so far been shown to be immunodominant. Only live F. tularensis affords effective host protection. Subcellular antigen preparations induce only a marginal protective response even when combined with potent adjuvants such as immunostimulating complexes (ISCOMs). In mice, intradermal injection of live F. tularensis but not of killed bacteria results in an early cytokine expression in the infected liver, including interleukin-12, tumor necrosis factor-α, and interferon-γ. This cytokine response seems to be a prerequisite for effective priming of T cells to an array of proteins of F. tularensis to occur.     

食物限制对黑线仓鼠免疫功能的影响

温带地区小型哺乳动物经常面临着食物资源的波动。食物对动物的免疫功能具有重要影响。将19只成年雄性黑线仓鼠(Cricetulus barabensis)随机分为自由取食组(n=9)和限食组(n=10)。注射植物血球凝集素(PHA)来测定细胞介导的免疫反应,用匙孔血蓝蛋白(KLH)免疫动物,然后测定抗KLH抗体的浓度以反映其体液免疫功能。旨在检验食物限制是否会抑制黑线仓鼠的细胞免疫和体液免疫功能。结果发现,与对照组相比,限食组黑线仓鼠具有较低的体重、体脂、脾脏鲜重、血清瘦素水平、免疫球蛋白Ig G和Ig M浓度。而限食对胸腺鲜重、白细胞数、皮质酮水平以及PHA反应没有显著影响。结果表明黑线仓鼠免疫系统的不同成分对限食反应存在差异,在食物资源短缺时,黑线仓鼠防御细胞外病原体的能力降低,从而导致生存能力的下降。