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Tenacibaculum sp., the causative bacterium of spotting disease of sea urchin Strongylocentroutus intermedius, has been known to enter into the viable but non-culturable (VBNC) state in 75% artificial seawater (ASW) at 5°C. From this fact it is suspected that this bacterium probably resuscitates from the VBNC state in summer. To examine whether non-culturable cells of Tenacibaculum sp. are able to regain the culturable state, VBNC cells in 75% ASW and 75% ASW containing homogenated sea urchin (HSU) were treated with temperature upshift (5°C to 25°C) and addition of iron chloride. Resuscitation from VBNC cells with temperature upshift was observed only in 75% ASW containing HSU for 3 days incubation after entry into the VBNC state. Resuscitation of VBNC cells treated with iron chloride was observed in each microcosm. Resuscitation from the VBNC state was confirmed by dilution experiments and measurements of generation time. Resuscitated cells regained original characteristics, agglutination against anti- Tenacibaculum sp. strain F-2, pathogenicity to sea urchin and morphology. [ABSTRACT FROM AUTHOR]

Background: Tumour necrosis factor-alpha (TNFα) is an essential regulator of immune responses and is implicated to relate to several types of disease susceptibilities. Population information on polymorphisms is essential for the study of genetic diseases. Aim: To obtain accurate information about single nucleotide polymorphisms (SNPs) in the TNFα gene in the Japanese population. Subjects and Methods: The entire TNFα gene was screened for SNPs by directly sequencing 48 chromosomes derived from 24 unrelated Japanese individuals. Allele frequencies of each polymorphism were determined and compared with those previously reported in other populations. Results: Three SNPs, -308G/A at nt -308, IVS1 + 125G/A at nt 492 and IVS3 + 104G/A at nt 1359 were observed, of which one (IVS3 + 104G/A at nt 1359) was novel. In addition, allele frequencies of -308G/A were remarkably different from those presented in the NCBI dbSNP, indicating a significant ethnic difference. Conclusions: The polymorphisms and allele frequencies obtained in this study will be useful for genetic studies of common diseases such as osteoporosis and rheumatoid arthritis in the Japanese population. [ABSTRACT FROM AUTHOR]

Background: Tumour necrosis factor alpha (TNFα) has come to be regarded as a potential osteoporotic factor, because it has stimulatory effects on cells of the osteoclast lineage and has been implicated in the pathogenesis of bone loss associated with oestrogen deficiency. We recently described genetic linkage between the TNFα locus and human osteoporosis by sib-pair analysis. However, the molecular mechanism by which this locus regulates bone mineral density (BMD) remains unknown. Aim: We investigated whether the observed linkage reflects a sequence variation which might affect expression of the TNFα gene or alter the function of TNFα protein. Subjects and methods: We examined three single-nucleotide polymorphisms (SNPs) of the TNFα gene in a group of 390 postmenopausal Japanese women living in northern Japan. Minor-allele frequencies for the three SNPs (-1031C, -863A and -857T) in this population were 0.16, 0.13 and 0.20, respectively. Results: Among the three SNPs examined, we observed a significant correlation only between the presence of a T allele at nt -1031 and decreased BMD, by analysis of variance. Among the three genotypic groups at nt -1031, mean BMD values were significantly higher in the T-negative genotype (C/C homozygotes; mean SD = 0.342 ± 0.052 g cm[sup -2]), compared with T-positive genotypes (T/T homozygotes, 0.309 ± 0.062 g cm[sup -2]; p = 0.0253 and T/C heterozygotes, 0.305 ± 0.062 g cm[sup -2]; p = 0.0164). Conclusions: Given the lines of evidence from different genetic studies, we suggest that TNFα may play a role in pathogenesis of osteoporosis. [ABSTRACT FROM AUTHOR]

Interleukin 11 (IL11) is a member of the interleukin 6 (IL6)-related cytokine subfamily, which stimulates T cell-dependent development of immunoglobulin-producing B cells. IL11 is also an important paracrine regulator of bone metabolism that induces formation of osteoclasts. In the work reported here, we sequenced the entire IL11 structural gene of 48 alleles in a Japanese test population. These experiments identified ten single-nucleotide polymorphisms (SNPs) and determined their allelic frequencies. One polymorphism was identified upstream of exon 1, one in exon 3, four in intron 4 and four in the 3′ untranslated region (3′UTR) of exon 5. Based on the genotype data, we constructed six haplotypes in the tested population. Two-way comparisons of SNPs revealed two combinations in complete linkage disequilibrium, one with SNPs at nucleotide positions 2753, 3644, 5154, and 5568, and another with SNPs at positions 3686, 5141, and 5734. These results will be useful in disease-association studies where a contribution of the human IL11 gene has been suspected, especially in disorders affecting immune response and bone metabolism. [ABSTRACT FROM AUTHOR]

This study was performed to examine the relationship between knee extension loss and the length of time after injury. We also wanted to compare anterior laxity in anterior cruciate ligament-injured knees in the early and late stages of injury. Loss of knee extension was measured in 81 patients with anterior cruciate ligament injury using lateral radiography with the injured leg relaxed and elevated. Extension loss was defined as the difference in maximal extension angle between the injured and uninjured knees. Average loss of extension was 3.6 degrees in the 17 knees in which the anterior cruciate ligament had been torn 3 weeks or less before examination (the acute injury group) and 0.6 degrees in the 64 knees in which the anterior cruciate ligament had been torn more than 3 weeks before examination (the chronic injury group). The extension loss in the acutely injured knees was significantly greater compared with that in the uninjured knees and in the chronically injured knees. Arthrometric measurements using the KT-1000 arthrometer were reliable to diagnose an acute tear. There was no correlation between the degree of extension loss and arthrometric anterior knee laxity measurements. In 12 patients, the initial extension deficit in the early stage of injury significantly resolved with time, and manual maximum arthrometric measurements of anterior knee laxity improved spontaneously with time. [ABSTRACT FROM AUTHOR]

Leukemia inhibitory factor (LIF) is a pleiotropic cytokine implicated in various pathological conditions, such as rheumatoid arthritis and osteoporosis. Despite the possible importance of LIF as a therapeutic target, little is known about the bioregulation of the human LIF gene. We here sequenced the entire structure of the LIF gene of 48 alleles in the Japanese population. These experiments identified four single-nucleotide polymorphisms (SNPs) and determined their allelic frequencies from a 48-allele sequence in the Japanese population. All four SNPs found in the LIF gene were located within exon 3, that is, a C/T at nucleotide (nt) position 3951, a C/G at nt position 4376, an A/C at nt position 4442, and a G/A at nt position 5961 (nucleotide numbering starts from the ATG start codon). Based on the genotypic data, we constructed four major haplotypes in the tested population. Two-way comparisons of SNPs revealed complete linkage disequilibrium between SNPs at positions 3951, 4376, and 4442. These results may prove to be useful as genetic markers for population-based disease-association studies in osteoporosis. [ABSTRACT FROM AUTHOR]

Osteopontin (OPN) is one of the major noncollagenous bone matrix proteins produced by osteoblasts and osteoclasts. We systematically surveyed the entire structure of the OPN gene for single-nucleotide polymorphisms (SNPs) by directly sequencing 48 alleles derived from 24 unrelated Japanese individuals. We identified 13 SNPs in the OPN gene. Ten polymorphisms were identified in introns 1, 3, and 5; 2 in the coding region of exons 6 and 7; and 1 in the 3′ untranslated region of exon 7. Allele frequencies for some of the polymorphisms were significantly different from those reported in the United States National Center for Biotechnology Information (NCBI) dbSNP database. These polymorphisms will be useful in genetic studies to evaluate the role of OPN proteins in bone metabolism. [ABSTRACT FROM AUTHOR]