To grow Mycobacterium leprae in cultured mouse peritoneal macrophages, studies were made on 1) the purification of M. leprae from lepromatous nodules by trypsinization, 2) growth experiment of purified M. leprae in cu1tured macrophages by in vivo infection-in vitro cultivation technique and 3) the observation of pathological changes in sp1eens of mice induced by intraperitoneal inoculation of purified M. leprae. Results are summarized as follows. 1. A simple and effective procedure is described for purification of M. leprae from biopsied nodules of lepromatous leprosy patients by trypsinization and high speed centrifugation. The procedure resulted in a good yie1d of homogeneous preparation of M. leprae with a negligible contamination of tissue debris. 2. Significant decreases were observed in the numbers of acid-fast bacilli in cultured macrophages and of macrophages harboring acid-fast bacilli by the length of intervals between the time of intraperitoneal inoculation of purified M. leprae and the time of initiation of macrophage cultures. 3. Microscopic examination of stained preparations of macrophages cultured by in vivo infection-in vitro cultivation technique indicated that an apparent increase in the number of acid-fast bacilli in the macrophages occurred when the cultures made at 24 hours and 1 week after inoculation were maintained in vitro up to 2 months or more. 4. Pathological changes in the spleens of mice inoculated with purified M. leprae were of mainly degenerative nature in the red pulp. No multiplication of M. leprae was observed in the spleens of mice up to 5 months after inoculation.
Adolescent ; Adult ; Animal ; Cells, Cultured ; Female ; Human ; Leprosy/microbiology* ; Macrophages/microbiology* ; Male ; Mice ; Mycobacterium leprae/growth & development* ; Mycobacterium leprae/isolation & purification ; Peritoneum/microbiology*

The oral carriage rate of Candida in healthy humans ranges from 40% to 60%. However for a prolonged period, the oral candidal prevalence in humans was documented essentially using data from studies in the West as their prevalence in inhabitants in different regions of the world, including Asia was not known. Yet, recent reports from a number of studies indicate the quality, quantity and prevalence of oral yeasts differ between Asia and other regions for reason that are still unclear. This mini review on such data from Asian studies on oral carriage of Candida provides another intriguing facet of the behavior of this ubiquitous yeast.
Areca ; Asian Continental Ancestry Group ; Candida ; classification ; isolation & purification ; Cohort Studies ; Colony Count, Microbial ; HIV Infections ; microbiology ; Humans ; Leprosy ; microbiology ; Mouth ; microbiology

No abstract available.
Animal ; Child ; Clinical Trials ; Dapsone/therapeutic use* ; Haplorhini ; Human ; Leprosy/drug therapy* ; Mycobacterium leprae/growth & development ; Sciuridae/microbiology ; Time Factors

OBJECTIVEMultiple locus variable number-tandem repeat (VNTR) analysis (MLVA) had been proposed as a means of strain typing for tracking of source and studying the transmission chain of pathogens. However, empirical data for a defined population from scale and duration were lacking for studying the transmission chain of leprosy.

METHODSMLVA on 7 VNTR loci was applied to the strain typing on prevalent Mycobacterium leprae isolates collected from Qiubei county, Yunnan province during 2002-2006 in the study on the relationship between geographic distribution and genotypes of M. leprae. The strain typing, combined with conventional epidemiological investigation was performed to trace the transmission chain.

RESULTS(1) Phylogenetic analyses through application of PAUP 4.0, The M. leprae were grouped into A, B, C, D and E strains according to the allelic range 9, 11-13, 15-26 and > 26 on the GTA9 locus. The strains with 9 copies on GTA9 locus, was named A. (2) Genotypes of strains from the five multi-case families located at North and North-West parts were similar and belonged to A strains. VNTR patterns of intra-family were identical or similar but not identical inter-family. (3) Not only A cluster appeared higher proportion in total isolates but also distributes cluster, indicating ongoing transmission from recent findings.

CONCLUSIONVNTR strain typing was suitable to trace the short chain of transmission in both small area and intra-families. Multi-case families might constitute epidemic foci and source of M. leprae in villages, causing the predominant strain or cluster which tends to be those identified in multi-case families and resulted in the spreading of leprosy. A long-term study was underway to reveal whether A strain was predominant strain and to observe the evolution of M. leprae in this spatially and temporally defined endemic population.

Female ; Genotype ; Humans ; Leprosy ; microbiology ; Male ; Minisatellite Repeats ; genetics ; Molecular Epidemiology ; Mycobacterium leprae ; classification ; genetics ; Phylogeny ; Polymerase Chain Reaction

Acid-fast microorganisms were isolated from 240 soil samples collected at two areas, Hiroshima, Japan and Seoul, Korea. The biological and biochemical characteristics of the isolated mycobacteria were tested and compared with those of 36 reference mycobacteria Strains. The isolation rate and distribution of these mycobacterial species from soil were compared using three kinds of media with emphasis on the two methods of isolation between the different geographical areas. One Strain from each of the 10 species among atypical mycobacteria isolated from soil in both areas was inoculated into ddY mice and the pathogenicity compared with that of Mycobacterium tuberculosis H37Rv up to 6 weeks. Susceptibility of the reisolated acid-fast bacilli to antimycobacterial agents was tested in vitro. Antibody responses against various mycobacterial antigens were tested using lepromatous type and tuberculoid type patient sera by the agar gel immunodiffusion. 1) No significant differences in the distribution of acid-fast bacilli were observed between soil samples from the two regions. 2) Rapid growers were by far the most frequent acid-fast bacilli isolated while no photochromogens were isolated from these soil samples. In addition, a minimal number of fastidious mycobacteria were isolated but not cultivable in subcultures. 3) Some of these soil acid-fast bacilli were capable of inducing only transient bacteriological and pathologic changes in mouse organs. 4) Acid-fast bacilli reisolated from organs of these infected mice were, in general, found to be resistant to antimycobacterial agents. 5) M. scrofulaceum antigen showed a precipitation reaction in agar gel immunodiffusion with the highest number of sera from leprosy patients.
Animal ; Leprosy/immunology ; Mice ; Mycobacteria, Atypical/drug effects ; Mycobacteria, Atypical/isolation & purification* ; Mycobacteria, Atypical/pathogenicity ; Mycobacterium/isolation & purification* ; Mycobacterium Infections/pathology ; Soil Microbiology*

OBJECTIVETo understand the genotypic mapping of Mycobacterium leprae identified in China and to compare with those from other countries to select suitable alleles for epidemiological investigation in the transmission chain of leprosy.

METHODSVarious number of tandem repeat(VNTR) in genomic DNA of Mycobacterium leprae was used in the present genotyping study. 33 skin biopsies from Wenshan prefecture,Yunnan province and 17 from other parts of China were studied. DNA extracted from skin biopsies of leprosy patients was subjected to PCR followed by agarose gel analysis and DNA sequencing to determine the number of repeats.

RESULTSLoci GGT-5,12-5,21-3 and 23-3 were as highly homogenous as 100%; The homogeneity of loci AC-8, 18-8, 27-5 and rpoT were 97%, 94%, 97% and 85% respectively. Loci GTA-9, AC-9 and 6-7 showed significant allelic diversity in isolates and the diversity of GTA-9 in Mycobacterium leprae isolated from China was also different from those identified other countries. We had subjected loci GTA-9 and the ten loci to phylogenetic tree analysis respectively.

CONCLUSIONThe present study revealed that the genotype of Mycobacterium leprae identified from China was close to the strains from the Philippines and India although a few loci were somehow differentiate. Locus 12-5 manifested as only 3 copies in China whereas 4-5 copies predominating in other countries. 12-5 locus might serve as a useful marker to diffrentiate Chinese strains from those in other countries. However, further study on the diversity of GTA-9 was needed in China. The molecular typing of Mycobacterium leprae from different geographic areas might be useful in studying the transmission of leprosy.

Alleles ; China ; epidemiology ; DNA, Bacterial ; Genotype ; Humans ; Leprosy ; epidemiology ; transmission ; Molecular Epidemiology ; Mycobacterium leprae ; genetics ; isolation & purification ; Polymerase Chain Reaction ; Skin ; microbiology

Adult ; Female ; Heat-Shock Proteins ; genetics ; Humans ; Leprosy ; diagnosis ; genetics ; microbiology ; Male ; Middle Aged ; Mycobacterium leprae ; pathogenicity ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length ; Skin ; metabolism

Adult ; Aged ; Aged, 80 and over ; China ; epidemiology ; Female ; Helicobacter Infections ; epidemiology ; Helicobacter pylori ; Hospitalization ; Humans ; Inpatients ; statistics & numerical data ; Leprosy ; epidemiology ; microbiology ; Male ; Middle Aged

We reviewed dinical, histological and ultrastructural findings of 124 cases of sural nerve biopsy specimens to delineate the trends of peripheral nerve diseases in our institute. Eighty-one were men and 43 were women. We categorized them into five groups: specific diagnosis (66 cases, 53.2%), axonal degeneration type (47 cases, 37.9%), demyelinating type (4 cases, 3.2%), mixed axonal degeneration-demyelinating type (6 cases, 4.8%) and normal (1 case, 0.9%). Cases with specific diagnosis included 21 inflammatory demyelinating polyneuropathy (15 chronic inflammatory demyelinating polyradiculoneuropathy, 6 Guillain-Barre disease), 13 hereditary motor and sensory neuropathy (7 Charcot-Marie-Tooth type I, 6 Charcot-Marie-Tooth type II), 10 vasculitis, 6 toxic neuropathy, 4 leprosy, 3 diabetic neuropathy, 2 alcoholic neuropathy, 1 Fabry's disease and other specific diseases (5 cases). In our cases, the proportion of specific diagnoses was higher, while the proportion of demyelinating peripheral neuropathies and normal were lower than those of Western series. The results of this study indicate that 1) a dose clinicopathologic correlation is important to make a precise diagnosis of peripheral nerve biopsy, 2) Biopsy under strict indication may reduce unnecessary histologic examination, 3) There is no difference in disease pattern of peripheral neuropathy between Western people and Koreans.
Adult ; Biopsy ; Charcot-Marie-Tooth Disease/pathology ; Demyelinating Diseases/pathology ; Fabry Disease/pathology ; Female ; Hereditary Motor and Sensory Neuropathies/pathology ; Human ; Korea ; Leprosy/pathology ; Male ; Microscopy, Electron ; Nerve Fibers, Myelinated/pathology ; Peripheral Nerves/ultrastructure ; Peripheral Nerves/pathology ; Peripheral Nervous System Diseases/pathology* ; Peripheral Nervous System Diseases/microbiology ; Polyradiculoneuropathy, Chronic Inflammatory Demyelinating/pathology ; Sural Nerve/ultrastructure ; Sural Nerve/pathology*