Numerical classification of 280 strains of slowly growing mycobacteria. Proposal of Mycobacterium tuberculosis series, Mycobacterium avium series, and Mycobacterium nonchromogenicum series

Numerical classification of 280 strains of slowly growing mycobacteria was carried out by testing each strain for 76 characters. The following fourteen clusters were observed: 1. M. tuberculosis, M. bovis, M. africanum, and M. microti; 2. M. haemophilum; 3. M. ulcerans; 4. M. xenopi; 5. M. kansasii; 6. M. szulgai; 7. M. gordonae; 8. M gastri; 9. M. avium, M. intracellulare, M. scrofulaceum, and M. asiaticum; 10. M. marinum; 11. M. simiae; 12. M. nonchromogenicum, "M. novum," M. terrae, and M. triviale; 13 M. malmoense; 14. M. shimoidei. The clusters composed of M. tuberculosis, M. bovis, M. africanum, and M. microti, of M. avium, M. intracellulare, M. scrofulaceum, and M. asiaticum, and of M. nonchromogenicum, "M. novum," M. terrae, and M. triviale appeared to be reduced to a single species each. The names having priority for each species should be M. tuberculosis, M. avium, and M nonchromogenicum, respectively. However, the clusters may, in practice, be called the M. tuberculosis series (complex), the M. avium series (complex), and the M. nonchromogenicum series (complex). The type species of these series are M. tuberculosis, M. avium, and M. nonchromogenicum, respectively. These series were characterized in this study.     

Modulation of macrophage subtypes by IRF5 determines osteoclastogenic potential

Bone-resorbing osteoclasts are differentiated from macrophages (MΦ) by M-CSF and RANKL. MΦ can be mainly classified into M1 and M2 MΦ, which are proinflammatory and anti-inflammatory, respectively, but little is known about their osteoclastogenic potential. Here, we investigated the osteoclastogenic potential of MΦ subtypes. When the two MΦ subtypes were differentiated into osteoclasts using M-CSF and RANKL, M2 MΦ more potently differentiated into osteoclasts than M1 MΦ. M2 MΦ generated with IL-4 or IL-10 also showed enhanced osteoclast differentiation compared with M1 MΦ induced by IFN-γ and lipopolysaccharide. In addition, robust bone-resorptive capacity and giant actin rings, which are features of mature osteoclasts, were observed in M2, but not M1 MΦ, under the osteoclast differentiation condition. Osteoclast differentiation was significantly increased in CD206⁺ M2 MΦ but not in CD86⁺ M1 MΦ. Compared with M1 MΦ, c-Fms and RANK were highly expressed in M2 MΦ. Enhanced osteoclastogenesis of M2 MΦ was mediated through sustained ERK activation, followed by efficient c-Fos and NFATc1 induction. Notably, the osteoclastogenic potential of M1 MΦ converted into M2 MΦ by exposure to M-CSF was higher than that of M2 MΦ converted into M1 MΦ by exposure to GM-CSF. Silencing IRF5, which is responsible for M1 MΦ polarization, increased osteoclast differentiation by enhancing c-Fms expression and activation of ERK, c-Fos, CREB, and NFATc1, which was inhibited by overexpression of IRF5. Collectively, M2 MΦ are suggested to be more efficient osteoclast precursors than M1 MΦ because of the attenuated expression of IRF5.     

A new deltocephaline leafhopper genus,Melanetettix, from Melanesia (Hemiptera: Cicadellidae: Deltocephalinae)

Melanetettix gen. nov. is described from Melanesia for 22 new species of leafhoppers. The new species fall into four groups. New species described are M. bicolor, M. bifidus, M. capitatus, M. curvatus, M. delmoi, M. elongatus, M. maai, M. marginatus, M. ocellatus, M. pallidus, M. rhamphodes, M. roseus, M. semeraroae, M. sinuatus, M. aculeus, M. bifurcatus, M. truncatus, M. clavatus, M. ensiferus, M. nielsoni, M. rotundatus and M. alatus. An identification key to the new species is provided along with discussion of the affinities of Melanetettix, which is close to Scaphoideus Uhler.     

薄荷地上部分的非挥发性化学成分研究

薄荷属( Mentha Linn.)植物薄荷( Mentha haplocalyx Briq.)的干燥地上部分为常用中药材[1],其主要药用成分为挥发油[2-3],其非挥发性成分也具有重要的药理作用。为详细了解薄荷的非挥发性成分,作者对薄荷地上部分乙醇提取物的乙酸乙酯和正丁醇萃取物的组成成分进行了分析。     

基于毛髓质指数探讨甘肃鼢鼠、高原鼢鼠、秦岭鼢鼠的分类地位

目前甘肃鼢鼠Myospalax cansus、高原鼢鼠 M.baileyi、秦岭鼢鼠M.rufescens的分类地位一直存有争议,多涉及到与中华鼢鼠M.fontanierii的分类关系.本文分别从成体甘肃鼢鼠、高原鼢鼠、秦岭鼢鼠、中华鼢鼠标本的胡须、头部、背部、腹部、前肢取毛样,经清洗和处理后,在倒置显微镜下观察,用目镜测微尺分别测量和计算其5个部位毛发的毛髓质指数.结果表明:甘肃鼢鼠与中华鼢鼠除胡须髓质指数无显著性差异外,其它部位及各部位综合均有显著差异.高原鼢鼠与秦岭鼢鼠除前肢毛髓质指数无显著性差异外,其它部位及各部位综合均有显著差异;与中华鼢鼠除前肢毛髓质指数无显著差异,其它部位及各部位综合均有显著差异.秦岭鼢鼠除与甘肃鼢鼠的腹部及与中华鼢鼠、高原鼢鼠的前肢毛髓质指数无显著差异外,与其它鼢鼠其余部位及各部位综合均有显著性差异.综合考虑,本研究结果支持甘肃鼢鼠、高原鼢鼠、秦岭鼢鼠各自独立为种的观点.     

PCR with subsequent sequencing of the 16S gene rRNA in species identification of mycobacteria of the non-tuberculosis complex

One hundred of mycobacterium cultures were assayed by the method of PCR with subsequent sequencing of the 16S rRNA region. The below mycobacterium species were identified: M. tuberculosis complex (n = 55), M. avium (n = 17), M. intracellulare (n = 4), M. scrofaleceum (n = 2), M. kansasii - M. gastri (n = 3), M. gordonae (n = 3), M. ulcerans - M. marinum (n = 1), M. smegmatis (m = 2), M. fortuitum (n = 11), M. peregrinum (n = 1) and M. chelonae - M. abscessus (n = 1). The method enabled the differentiation of species M. avium from M. intracellulare and M. peregrinum from M. fortuitum, which could not be differentiated by using the classic biochemical and bacteriological methods. Genetic heterogeneity of the mycobacterium strains of M. avium, M. fortuitum and M. gordonae was also established by PCR plus sequencing of the 16S rRNA region.     

Revision of the subgenus Udamochiras of Melaloncha bee-killing flies (Diptera: Phoridae: Metopininae)

The genus Melaloncha Brues is defined and groundplan character states established based on outgroup comparison with Phalacrotophora Enderlein and Melittophora Brues. Major groupings within Melaloncha are recognized, and two subgenera are established: Udamochiras Enderlein (type species M. colossia (Enderlein)) and Melaloncha s.s. (type species M. pulchella Brues). Subgenus Udamochiras is revised and 42 species are recognized, including the following 33 new to science: M. anaticula , M. angustifrons , M. apicula , M. aprica , M. basella , M. biseta , M. brevicarina , M. carinata , M. compressicauda , M. exigua , M. falcata , M. flavilata , M. hamata , M. hansoni , M. horologia , M. individa , M. lobata , M. maculifrons , M. parkeri , M. paxilla , M. premordica , M. rhampha , M. rhypopoda , M. rostrata , M. sinuosa , M. spatula , M. spicula , M. triangularis , M. trua , M. valeria , M. vargasi , M. villosa , M. woodi . Melaloncha simillima Borgmeier is removed from synonymy with M. piliapex Borgmeier and reinstated as a separate species; lectotypes are designated for both.  © 2004 The Linnean Society of London, Zoological Journal of the Linnean Society , 2004, 140 , 1−42     

Occurrence of RD9 region and 500 bp fragment among clinical isolates of Mycobacterium tuberculosis and Mycobacterium bovis

The present investigation dealt with the identification of Mycobacterium tuberculosis and M. bovis by RD9 region and 500 bp fragment PCR assays. Eight M. tuberculosis and 5 M. bovis characterized and identified from 40 human sputum and 41 bovine lung specimens and 20 M. tuberculosis and 9 M. bovis strains maintained at Mycobacteria Laboratory, Indian Veterinary Research Institute were included in this study. In this way, 28 M. tuberculosis and 14 M. bovis strains and, for comparison and control purpose, M. tuberculosis H37Rv, M. bovis BCG, M. canetti, M. smegmatis, M. phlei, M. chelonae, M. kansasii, M. xenopi and M. avium were subjected to RD9 and 500 bp amplification by PCR. All M. tuberculosis strains, M. tuberculosis H37 Rv and M. canetti yielded a product of 333 bp which showed presence of RD9 region in these strains, whereas all M. bovis yielded a product of 206 bp with RD9 PCR assay. There was no ampli-fication product found in M. bovis BCG, M. xenopi, M. smegmatis, M. phlei, M. chelonae, M. kansasii, and M. avium. PCR based on 500 bp fragment showed a product of 500 bp in all M. bovis strains and M. bovis BCG. There was no amplification product of 500 bp found in M. canetti, M. smegmatis, M. phlei, M. chelonae, M. avium, M. kansasii, M. xenopi and was absent in all M. tuberculosis strains. The PCR assay results correlated 100% with the culture and biochemical results of the isolates. Our study suggested that PCR based on RD9 and 500 bp may effectively identify two closely related species of M. tuberculosis and M. bovis.     

Mechanisms of immune activation of human immunodeficiency virus in monocytes/macrophages.

Monocytes/macrophages (M/M) are the major host of human immunodeficiency virus (HIV) in solid tissues. However, blood monocytes are nonpermissive for HIV infection, indicating that M/M activation or differentiation is necessary for HIV replication. Since M/M are activated during immune responses, we investigated the effect of T-cell activation on HIV expression in M/M derived from peripheral blood of HIV-infected individuals. Previously, we reported that coculture of monocytes from HIV-infected donors with T cells and mitogens resulted in M/M differentiation and HIV expression. Production of HIV by M/M from infected donors required direct contact between monocytes and T cells (for the first 24 h), and the response to alloantigens, but not mitogens, was restricted to HLA-DR. In this study, we found that HIV was more readily recovered from M/M of asymptomatic HIV seropositive donors (69%) than from M/M of symptomatic donors (57%). Viral antigens (e.g., inactivated herpes simplex virus) could initiate the immune response and HIV expression. The ability of noninfected T cells to activate HIV expression in M/M and observations that treatments of M/M with antibodies to deplete T cells did not reduce HIV expression suggested that the monocytes were endogenously infected. To define the aspects of immune activation specifically involved in initiating HIV expression in M/M, interactions of M/M and T cells and participation of cytokines were investigated. The T cell which activated M/M was CD4+ CD8-. Fixed allogeneic cells are known to induce T-cell activation but were not able to serve as antigen for M/M differentiation, suggesting that M/M may need to function as antigen-presenting cells to receive the signal to differentiate and express HIV. Blocking of M/M-T-cell interaction with antibodies directed against LFA-1 or interleukin-1 prevented HIV expression. However, inhibition of later stages of T-cell activation, such as blocking of interleukin-2 receptors, did not diminish HIV expression in M/M. Consistent with the requirement for cell-cell contact between M/M and T cells, a variety of cytokines were unable to initiate HIV replication in M/M. The ability of T cells to induce cellular differentiation and HIV replication in M/M in vitro suggests that initiation of an immune response to an antigen, such as an opportunistic pathogen, could be a mechanism by which HIV disseminates to tissues in vivo.     

Towards a phylogeny and definition of species at the molecular level within the genus Mycobacterium

16S rRNA sequences from Mycobacterium tuberculosis, M. avium, M. gastri, M. kansasii, M. marinum, M. chelonae, M. smegmatis, M. terrae, M. gordonae, M. scrofulaceum, M. szulgai, M. intracellulare, M. nonchromogenicum, M. xenopi, M. malmoense, M. simiae, M. flavescens, M. fortuitum, and M. paratuberculosis were determined and compared. The sequence data were used to infer a phylogenetic tree, which provided the basis for a systematic phylogenetic analysis of the genus Mycobacterium. The groups of slow- and fast-growing mycobacteria could be differentiated as distinct entities. We found that M. simiae occupies phylogenetically an intermediate position between these two groups. The phylogenetic relatedness within the slow-growing species did not reflect the Runyon classification of photochromogenic, scotchromogenic, and nonchromogenic mycobacteria. In general, the phylogenetic units identified by using rRNA sequences confirmed the validity of phenotypically defined species; an exception was M. gastri, which was indistinguishable from M. kansasii when this kind of analysis was used.     

AFLP analysis of the genetic diversity of Meloidogyne chitwoodi and M. fallax, major agricultural pests

M. chitwoodi and M. fallax populations are clustered and separated from the other species studied. The genetic diversity observed for M. incognita, M. arenaria, M. javanica, M. hapla, and M. mayaguensis correlates well with the previously validated species. Two main groups can be identified within the M. chitwoodi/M. fallax cluster, the first group comprises only M. chitwoodi populations whereas the second group is made of M. chitwoodi and M. fallax populations. Moreover, M. chitwoodi displays a higher genetic diversity than M. fallax and is characterised by the presence of several clusters.     

Host-parasite associations of Eimeria spp. (Apicomplexa:Eimeriidae) in kangaroos and wallabies of the genus Macropus (Marsupialia:Macropodidae)

Faecal samples from 514 kangaroos and wallabies representing 12 species of the genus Macropus were examined for oocysts of Eimeria spp. Six species of Eimeria were redescribed from their type hosts, and on the basis of finding homologous oocysts in the faeces of other Macropus spp., host ranges for these coccidia were extended. Eimeria hestermani Mykytowycz, 1964 is redescribed from M. giganteus (eastern grey kangaroo) and is described from M. fuliginosus (western grey kangaroo), M. rufogriseus (red-necked wallaby), M. dorsalis (black-striped wallaby), and M. eugenii (tammar wallaby). E. toganmainensis Mykytowycz, 1964 is redescribed from M. rufus (red kangaroo) and the host range is extended to M. giganteus, M. fuliginosus, M. rufogriseus and M. eugenii. E. wilcanniensis Mykytowycz, 1964 is redescribed from M. rufus, and the host range is extended to M. giganteus, M. fuliginosus and M. robustus (euro or wallaroo). E. macropodis Wenyon & Scott, 1925 is redescribed from M. rufogriseus, and is described from M. giganteus, M. fuliginosus, M. rufus, M. irma (western brush wallaby), M. parryi (whip-tailed wallaby), M. dorsalis, M. eugenii, and M. parma (parma wallaby). E. fausti Yakimoff & Matschoulsky, 1936, E. cunnamullensis Mykytowycz, 1964 and E. purchasei Mykytowycz, 1964 are synonymized with E. macropodis. E. marsupialium Yakimoff & Matschoulsky, 1936 is redescribed from M. giganteus, and from M. fuliginosus. E. gungahlinensis Mykytowycz, 1964 is redescribed from M. fuliginosus, and from M. giganteus. Seven new species of Eimeria are described. E. flindersi, new species, is described from M. eugenii, M. rufogriseus, and M. antilopinus (antilopine wallaroo). E. prionotemni, new species, is described from M. eugenii, M. parryi, M. rufogriseus, M. agilis (agile wallaby) and M. dorsalis. E. mykytowyczi, new species, is described from M. agilis, M. antilopinus, and M. parryi. E. parryi, new species, is described from M. parryi. E. yathongensis, new species, is described from M. fuliginosus and M. giganteus. E. parma, new species, is described from M. parma, and E. desmaresti, new species, is described from M. rufogriseus. E. kogoni Mykytowycz, 1964, and E. rufusi Prasad, 1960 are considered species inquirendae. The host-parasite associations of these coccidia, and of similar species of Eimeria in other genera of Macropodoid marsupials, are discussed in relation to the postulated phylogeny of the hosts.     

DNA hybridization analysis of mycobacterial DNA using the 18-kDa protein gene of Mycobacterium leprae

Abstract DNA hybridization studies using a 611-base pair (bp) probe, encoding the entire 18-kDa protein of Mycobacterium leprae , demonstrated that M. simiae, M. intracellulare, M. kansasii, M. terrae , ADM-2, M. avium, M. scrofulaceum, M. gordonae and M. chelonei appear to posses DNA sequences homologous to the 18-kDa protein gene of M. leprae . RFLP analysis revealed that the restriction sites in the M. leprae 18-kDa gene were not conserved in the putative gene homologs of M. simiae and M. intracellulare . The restriction patterns observed with the 611-bp probe were useful in differentiating M. intracellulare, M. simiae , and M. leprae from each other, as well as in distinguishing strains of M. simiae serovar 1. Finally, the presence of homologous sequences in various mycobacteria did not affect the specificity of a previously described PCR test for detection of M. leprae , based on the M. leprae 18-kDa protein gene.     

麂属(Muntiacus)动物线粒体12S rRNA、细胞色素b基因和多药耐药基因序列分析及其分子进化关系

对麂属（Muntiacus)中的3种动物：赤麂（M.muntjak)(2n=6♀,7♂）,小麂（M.reevesi)(2n=46),黑麂（M.cri.nifrons)(2n=8♀,9♂）线粒体DNA 12S rRNA和细胞色素b基因784bp左右的片段和核基因－多药耐药基因（multidrug resistance,MDR1)的728bp左右的片段进行了序列分析,并根据序列信息建立了分子系统树,同时探讨了这3种动物的起源,分类地位及进化关系。     

Relationships between rust resistance genes at the M locus in flax

Genes at the M locus in flax ( Linum usitatissimum ) that confer resistance to flax rust ( Melampsora lini ) occur in complex haplotypes containing up to 15 related genes or gene fragments. We have cloned two additional functional resistance genes at this locus, M1 and M3 , by transposon tagging and candidate gene approaches, and investigated the genetic relationships between four genes ( M , M1 , M3 and M4 ) by recombination analysis. M1 and M3 , like M , are members of the nucleotide binding site, leucine-rich repeat (NBS-LRR) family. Comparisons of the predicted M1 and M3 amino acid sequences with M and L6 reveal that: (i) M1 contains four additional LRRs, probably as a result of an unequal crossover event between duplicated regions; (ii) M1 shares large segments of exact identity with M and M3, indicative of intragenic recombination events; and (iii) a large number of amino acid differences are scattered throughout the M, M1 and M3 proteins. Recombination analysis (here and in previous studies) has revealed that M readily recombines with M1 , M3 and M4 , whereas these three genes fail to recombine despite large family sizes (>5800) in two test-cross families, suggesting that they may occupy allelic positions in the gene cluster. Several restriction fragment length polymorphism markers within or near the M locus were mapped with respect to seven crossover events between M and M1 . The results of this and previous studies provide evidence of structural differences between: (i) homoeologous loci in the different genomes of flax; (ii) different haplotypes at the M locus; (iii) different resistance genes in the M group; and (iv) the flanking regions downstream of M locus resistance genes.     

Biotrophic fungi from Kenya: Ten new species and some new records of Meliolaceae

As part of a research programme studying the diversity of fungi associated with rare plants in Kenya, nine new species of Meliola and one of Cryptomeliola associated with seven plant families are described and illustrated. They include Meliola craterispermi, M. dendroseta, M. fragrans, M. kakamegensis, M. longiappressoriata, M. muhakae, M. phaeomaculans, M. taitensis, M. trilepisii and C. Natans. Host families include Araliaceae, Ebenaceae, Euphorbiaceae, Loganiaceae, Melastomataceae, Menispermaceae, Moraceae and Rubiaceae. New records for Kenya include Meliola ambigua, M. artabotrydis, M. carissae, M. clerodendri, M. impatientis, M. pycnostachydis, M. phytolaccae and M. polytricha.     

Pyruvate kinase isozymes in man

Anti human M2 type and anti human L type pyruvate kinase sera allowed us to distinguish two groups of pyruvate kinase in man. Erythrocyte and liver (L type) enzymes on the one hand were inhibited by anti L and not all by anti M2 serum; pyruvate kinase from all the other tissues on the other hand were inhibited by anti M2 and not at all by anti L serum. This latter group represent the M type pyruvate kinase isozymes. The M type isozymes have been studied by electrofocusing in thin layer acrylamide-ampholine gel. In adult tissues 4 types of isozymes were found, designated, from acid to alkaline pH, as M2 (predominant form in spleen, leukocytes, lung...), M3, M4 and M1 (predominant form in muscle and brain). In foetal tissues an extra band M2, called M2f, more anodic than M2, was added to the previously described isozymes. Except in brain (in which the isozymes M2, M3, M4 and M1 were found), the most anodic bands (M2f, M2 and M3) were predominant in all the foetal tissues. The isozymes M2f and M2 seem therefore to be the original M type pyruvate kinase forms from which the other isozymes issue. The rate of each isozyme seems to depend on tissue factors characterizing the state of differentiation of some tissues, as indicated by the ability of adult muscle extracts to change the isozymes M2 and M3 into more cathodic forms.     

Distribution of phthiocerol diester, phenolic mycosides and related compounds in mycobacteria

Among 28 mycobacterial species studied, only Mycobacterium tuberculosis, M. bovis, M. africanum, M. marinum, M. kansasii, M. gastri and M. ulcerans produced waxes yielding long-chain beta-diol components (called phthiocerol and companions) and polymethyl-branched fatty acids on saponification. The same mycobacterial species also produced diesters of phenol phthiocerol and companions. Fatty acids esterifying these fatty alcohols in M. marinum and M. ulcerans were found to belong to the phthioceranic series (dextrorotatory fatty acids), in contrast to those of the other species (laevorotatory fatty acids called mycocerosic acids), both groups having the same chain length and methyl-branched positions. M. kansasii and M. gastri contained the same waxes with identical structures, as did M. tuberculosis, M. bovis and M. africanum. Neither the type strain of M. tuberculosis, nor that of M. bovis or M. marinum accumulated the strain-specific phenolic glycolipids.