Production of parathyroid hormone-related peptide in a patient with acute lymphocytic leukemia with extensive osteolytic lesions and hypercalcemia.
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Adolescent↗
Biomedical subjects
F Taguchi
Publications and source records attributed to F Taguchi.
Neurovirulence for rats of the JHMV variants escaped from neutralization with the S1-specific monoclonal antibodies.
We have studied the neurovirulence for rats of the MAb-resistant variants isolated from a highly neurovirulent JHMV, cl-2. The variants, MM6 and MM13, with point mutation located within the N terminal 100 amino acids (aa) of the S1 protein showed no alteration in neurovirulence in comparison with cl-2, showing high neurovirulence. The variants, MM65 and MM85, with a deletion composed of about 150 aa located in the middle of the S1 subunit were revealed to be non-neurovirulent. A variant MM78 with one aa deletion, asparagic acid at number 543 from the N terminus of the S1, was shown to be low-virulence. The neurovirulence of these viruses paralleled with the viral growth potential in the rat brain. However, all of these variants as well as parental cl-2 showed high neurovirulence for mice. These results suggest that the domain composed of about 150 aa in the middle of the S1 is critical for high-neurovirulence of JHMV for rats.
Animals↗
Localization of neutralizing epitopes and receptor-binding site in murine coronavirus spike protein.
To identify the localization of the epitopes recognized by monoclonal antibodies (MAbs) against the S1 subunit of the murine coronavirus JHMV spike protein, we have expressed the S1 proteins with different deletions from the C terminus of the S1. All of MAbs in groups A and B recognized the S1N(330) composed of 330 amino acids (aa) from the N terminus of the S1 and the larger S1 deletion mutants, but failed to react with the S1N(220) composed of 220 aa. MAbs in group C reacted only with the S1utt protein without any deletion. These results indicated that the S1N330 comprised the cluster of epitopes recognized by MAbs in groups A and B. These results together with the fact that all the MAbs in group B retained the high neutralizing activity suggested that the N terminus 330 aa are responsible for binding to the MHV-specific receptors. In pursuit of this possibility, we have expressed the receptor protein and examined the binding of each S1 deletion mutants to the receptor. It was demonstrated that the S1N(330) protein as well as other S1 deletion mutants larger than S1N(330) bound to the receptor. These results indicated that a domain composed of 330 aa at the N terminus of the S1 protein is responsible for binding to the MHV-specific receptor.
Animals↗
Rapid detection of Vibrio cholerae with a new selective enrichment medium and polymerase chain reaction.
The inhibitory effect of metallic EDTA compounds on growth of Vibrio cholerae and Escherichia coli was studied. Only Fe-EDTA among the compounds tested showed pH-dependent growth inhibition on E. coli at pH 9.0, but no inhibition of V. cholerae at the same pH. By addition of Fe-EDTA as a selective inhibitor, a novel enrichment broth (tentatively designated as VCF broth) for the selective isolation and cultivation of V. cholerae from other Gram-negative bacilli has been developed, and the selective enrichment capacity of VCF broth for V. cholerae and selective inhibiting activity against E. coli were significantly higher than those of alkaline peptone water. A simple procedure for rapid detection of V. cholerae by selective enrichment for 6 hr with VCF broth and then amplification of the cholera toxin target DNA fragment by the polymerase chain reaction was presented. VCF broth may be a useful tool for the selective enrichment of V. cholerae in bacterial examinations.
Bacteriological Techniques↗
Role of C5a-ase in group B streptococcal resistance to opsonophagocytic killing.
Type III group B streptococci (GBS) can be subdivided into three subtypes, RDP III-1, III-2, and III-3, on the basis of numerical analysis of HindIII restriction endonuclease digestion patterns (HindIII RDP) with their chromosomal DNAs. In the present study, the effect of C5a on opsonophagocytic killing of a representative strain from each RDP type was investigated by using a novel optical method for determining opsonophagocytic killing, and the effect of C5a-ase treatment of C5a on opsonophagocytic killing was also investigated. Pre-stimulation of polymorphonuclear leukocytes (PMNs) with C5a significantly increased opsonophagocytic killing of all three strains. The increase in killing was abolished by pretreating the C5a with GBS that express C5a-ase, a treatment that also destroyed the chemoattractant activity of the C5a. The kinetics of killing of the RDP III-2 strain differed from those of the other two strains. The survival of the RDP III-2 bacteria continued to decline over the entire 60-min incubation of the opsonophagocytic assay when PMNs were prestimulated with C5a or with C5a that had been inactivated with GBS C5a-ase (dC5a). In contrast, killing of the RDP III-1 and III-3 strains almost ceased after 20 or 60 min when PMNs were prestimulated with dC5a or C5a, respectively. A difference in bacterial killing between the III-2 strain and the III-1 and III-3 strains therefore became increasingly apparent with prolonged incubation time. The percentage of bacteria surviving in the extracellular fluid was approximately the same as the percentages of bacteria surviving in both intracellular and extracellular locations when PMNs were prestimulated with either C5a or dC5a. These data imply that the majority of bacterial killing occurred following phagocytosis and suggest that the enhanced killing of GBS following prestimulation of PMNs with C5a resulted from increased ingestion of the bacteria.
Adhesins, Bacterial↗
Parent-to-child transmission is relatively common in the spread of the human polyomavirus JC virus.
JC polyomavirus (JCV), the causative agent of progressive multifocal leukoencephalopathy, is ubiquitous in the human population, infecting children asymptomatically and then persisting in renal tissue. In most adults, renal JCV replicates and the progeny are excreted in urine. We used this urinary JCV to elucidate the routes of JCV transmission. A 610-bp JCV DNA region (IG region) encompassing the 3'-terminal sequences of both T-antigen and VP1 (major capsid protein) genes was amplified by means of PCR from urine specimens collected from all members of seven families. JCV strains were then unequivocally identified by the nucleotide sequences of the amplified IG regions. We could identify 18 distinctive JCV strains from 27 individuals. Different JCV strains were detected from all unrelated persons. However, the same viral strain was detected from one (four families), two (one family), or three offspring (one family) as well as from the fathers (three families) or from the mothers (three families). In total, the JCV strains detected in half of the JCV-positive children were identified in their parents. Since most humans are infected during childhood, these findings indicated that JCV is transmitted frequently from parents to children. We roughly estimated that 50% of JCV transmission occurs by this route and that the other 50% occurs outside the family.
Adult↗
The S2 subunit of the murine coronavirus spike protein is not involved in receptor binding.
The receptor-binding capacity of the S2 subunit of the murine coronavirus S protein was examined by testing the inhibition of virus-receptor binding. Sp-4 virus and S1N(330), which consists of the N-terminal 330 amino acids of the S1 protein, both of which exhibited receptor-binding capacity, were able to prevent the binding of cl-2 virus to the receptor, while the mutant protein S1N(330)-159, which failed to bind to the receptor protein, and S2TM-, which lacks the transmembrane and cytoplasmic domains normally existing in the S2, were unable to prevent the binding of cl-2. By using cultured DBT cells, it was revealed that the infection of cells by cl-2 virus was significantly inhibited by S1N(330) but not by S2TM-. These results indicate that the S2 protein is not involved in the receptor binding of murine coronaviruses.
Animals↗
[Analysis of duodenal bile acids in patients with colorectal tumors and control subjects].
Bile acids are believed to play a role in the etiology of colorectal cancer. To investigate a possible relationship between bile acids and colorectal tumors, duodenal bile acids were analyzed in 18 patients with colorectal adenomas, 18 patients with colorectal carcinoma and 18 control subjects. Using high performance liquid chromatography and immobilized 3 alpha-hydroxysteroid dehydrogenase in column form, significant increases in the proportion of chenodeoxycholic acid and significant decreases in the proportion of deoxycholic acid and lithocholic acid were found in the bile of patients with colorectal adenoma or carcinoma compared with the control subjects. The data support the concept that bile acids have a role to play in the development of large bowel tumors.
Adenoma↗
[Influenza-bacillus: who discovered the organism?].
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Animals↗
Alteration in the JC polyomavirus genome is enhanced in immunosuppressed renal transplant patients.
JC polyomavirus (JCV) DNAs derived from urine generally contain an archetypal regulatory sequence, whereas regulatory regions of JCVs in the brain with progressive multifocal leukoencephalopathy (PML) have specifically altered regulatory regions. However, JCV DNAs with regulatory regions that deviate from the archetype sometimes occur in the urine of non-PML patients. In this study, we compared the incidence of these rearranged regulatory regions between immunosuppressed (renal transplant recipients) and nonimmunosuppressed patients. We found that the incidence of rearranged JCV regulatory regions was significantly higher in renal transplant patients than that in nonimmunosuppressed patients. This finding suggests that immunosuppression would permit accumulation of JCV with rearranged regulatory regions. On the other hand, from the following observations, we conclude that rearranged regulatory regions unique to PML-type JCVs rarely occur even in renal transplant patients: (1) rearrangements in JCVs from these patients were almost exclusively simple, that is deletions or duplications; (2) duplication of domain A or deletion of domain B, each being a feature common to most PML-type regulatory regions, was rarely detected in JCVs from the patients. The data are consistent with the fact that PML is a rare disease in patients with decreased immune competence and support the hypothesis that changes in the JCV regulatory region may be involved with the etiology of PML.
Adult↗
A murine coronavirus MHV-S isolate from persistently infected cells has a leader and two consensus sequences between the M and N genes.
A plaque-cloned mouse hepatitis virus mutant, MHV-S No. 8, was isolated from Ki-BALB cells persistently infected with MHV-S. The mRNAs 1 to 6 were larger in the mutant, whereas there was no difference between the two viruses in the size of the smallest mRNA, mRNA 7. Sequence analyses of the genomic RNA, mRNA 6, and mRNA 7 of the two viruses revealed that an additional 111 nt were inserted just upstream of the intergenic consensus sequence preceding the N gene in MHV-S No. 8. The inserted region consisted of two different parts; the 3'-most 30 nt corresponded to nucleotides 28 to 57 of the leader sequence and the 5'-most 81 nt corresponded to nucleotides 58 to 138 of mRNA 7. This structure of No. 8 was most likely generated by RNA-RNA recombination between genomic RNA and subgenomic RNA species. The nucleotide insertion in the intergenic sequence between genes M and N resulted in two consensus sequences separated by 111 nt. Primer extension analysis revealed that the amount of a slightly larger, subgenomic mRNA resulting from initiation of synthesis at the upstream consensus sequence was only 5% of the usual sized mRNA 7 initiated from the downstream consensus sequence.
Animals↗
Clinical significance of serum iron and ferritin in patients with colorectal cancer.
To clarify the significance of serum iron and ferritin as indicators of iron loss caused by continuous bleeding, and, thus, to determine their value as markers of colorectal cancer, values for the two were compared in male patients with early and advanced colorectal cancer and age-matched male controls. The mean value of serum iron levels in patients with advanced colorectal cancer was significantly decreased compared with values in patients with early colorectal cancer and controls, 50.5 +/- 38.6 micrograms/dl vs 93.0 +/- 32.1 micrograms/dl and 107.1 +/- 32.9 micrograms/dl, respectively (p < 0.001). The mean value of serum ferritin levels in patients with early and advanced colorectal cancer was also significantly decreased compared with controls, 80.5 +/- 35.0 ng/ml (p < 0.01) and 48.8 +/- 72.8 ng/ml (p < 0.001), respectively, vs 117.1 +/- 46.8 ng/ml. However, there was no significant difference between mean serum iron levels in patients with early colorectal cancer and controls. Eighteen (78.3%) of the 23 patients with advanced colorectal cancer and 3 (16.7%) of the 18 patients with early colorectal cancer had serum iron levels below 85 micrograms/dl and serum ferritin levels below 60 ng/ml. Levels of both serum iron and ferritin, without clinically evident anemia, are useful indicators of advanced colorectal cancer.
Aged↗
Occurrence of multiple JC virus variants with distinctive regulatory sequences in the brain of a single patient with progressive multifocal leukoencephalopathy.
We established 99 JC virus (JCV) DNA clones directly from the brain of a single patient with progressive multifocal leukoencephalopathy (PML). Based upon restriction patterns, the cloned viral DNAs were classified into two major groups (NY-1A and -1B) containing 53 and 35 clones, respectively, and several minor groups containing one or a few clones. The regulatory sequences of representative clones were compared with the archetypal regulatory sequence, which has been detected in JCV DNAs cloned from the urine of healthy and nonimmunosuppressed individuals. The regulatory sequence of NY-1B had the two structural features common to most PML-type regulatory regions, duplication and deletion of specific segments in the archetypal sequence, while that of NY-1A contained a small deletion and an insertion of a 29-bp sequence originating from the early region of the JCV genome. A regulatory region similar to that of NY-1A has never been detected in JCV isolates obtained thus far.
AIDS-Related Opportunistic Infections↗
[Bacterial contamination of hair washing liquids].
To determine the extent of contamination by bacteria of hair-washing shampoo and rinse used professionally at barber shops and hair-dressing saloons, quantitative isolation of bacteria were performed by using a total of 39 samples of shampoo and rinse fluid obtained from 17 facilities. It was found that a maximal number of 1 x 10(7)/ml colony forming units/ml of bacteria were isolated from 60.7% (17 out of 28 samples) of the shampoo and 45.5% (5 out of 11) of the rinse. Gram-negative bacilli were the predominant strains (87.9%) involved in bacterial contamination and the major isolates were Serratia marcescens (43.3%, most frequently isolated), Pseudomonas cepacia, P. fluorescens, P. aeruginosa and Klebsiella pneumoniae, all which are widely recognized as nosocomial-infection causing pathogens. These results indicate hair-washing liquids for professional use are contaminated with a great number of gram-negative bacteria, being possible causes of nosocomial infections, and much attention should be paid to the sanitation and cleanliness of the shampoo and rinse for hair-washing.
Bacteria↗
Localization of neutralizing epitopes and the receptor-binding site within the amino-terminal 330 amino acids of the murine coronavirus spike protein.
To localize the epitopes recognized by monoclonal antibodies (MAbs) specific for the S1 subunit of the murine coronavirus JHMV spike protein, we have expressed S1 proteins with different deletions from the C terminus of S1. S1utt is composed of the entire 769-amino-acid (aa) S1 protein; S1NM, S1N, S1n(330), and S1n(220) are deletion mutants with 594, 453, 330, and 220 aa from the N terminus of the S1 protein. The expressed S1 deletion mutant proteins were examined for reactivities to a panel of MAbs. All MAbs classified in groups A and B, those reactive to most mouse hepatitis virus (MHV) strains and those specific for isolate JHMV, respectively, recognized S1N(330) and the larger S1 deletion mutants but failed to react with S1N(220). MAbs in group C, specific for the larger S protein of JHMV, reacted only with the S1utt protein without any deletion. These results indicated that the domain composed of the N-terminal 330 aa comprised the cluster of conformational epitopes recognized by MAbs in groups A and B. It was also shown that the epitopes of MAbs in group C were not restricted to the region missing in the smaller S protein. These results together with the fact that all MAbs in group B retained high neutralizing activity suggested the possibility that the N-terminal 330 aa are responsible for binding to the MHV-specific receptors. In investigate this possibility, we expressed the receptor protein and examined the binding of each S1 deletion mutant to the receptor. It was demonstrated that the S1N(330) protein as well as other S1 deletion mutants larger than S1N(330) bound to the receptor. These results indicated that a domain composed of 330 aa at the N terminus of the S1 protein is responsible for binding to the MHV-specific receptor.
Antibodies, Monoclonal↗
[A simple method for differentiation of hydrogen sulfide-producing bacteria by the pH-dependent EDTA-sensitivity test].
The pH-dependent EDTA-sensitivity test was performed to differentiate several strains of bacteria forming black colonies by the production of hydrogen sulfide on TCBS (thiosulfate-citrate-bile salt-sucrose agar) medium (tentatively designated as hydrogen-sulfide production bacteria). Two halotorelant strains of 16 hydrogen sulfide-producing strains showed the same bacteriological properties and isoprenoid quinone type as did a reference strain of Proteus mirabilis and were classified into the EDTA-insensitive group as were P. mirabilis and P. vulgaris. On the other hand, the other 14 halophilic strains, showing similar but not identical bacteriological properties or the isoprenoid quinone type to those of Shewanella putrefaciens IFO 3908, were classified into the "EDTA-sensitive (at pH 5)" group as were some species of the genus Vibrio. By the same sensitivity test, S. putrefaciens IFO 3908 was classified into the "EDTA-sensitive (at any pH)" group. These results indicate that the pH-dependent EDTA-sensitivity test is useful for differentiation of bacterial isolates producing hydrogen sulfide and having similar bacteriological properties.
Bacteria↗
Phylogenetic comparison between archetypal and disease-associated JC virus isolates in Japan.
We examined the phylogenetical correlation between two types of JC virus (JCV) isolates, archetypes derived from the urine of nonimmunocompromised individuals and PML-types derived from the brain of patients with progressive multifocal leukoencephalopathy (PML) in Japan. A phylogenetic tree was constructed for eight JCV isolates, five PML-types obtained in this and previous studies and three representative archetypes, from DNA sequence data on the VP1 (major capsid protein) gene. The eight isolates were divided into two major groups, named subtypes MY and CY after the representative archetypal isolates. Four of five PML-type isolates belonged to subtype MY, and the other one to subtype CY. Isolates belonging to subtype MY were further divided into two groups; one group containing archetype MY and three PML-types and the other one containing archetype YI and a PML-type. These findings, together with those in our previous study that correlated various JCV isolates in the world provide evidence for the hypothesis that JCVs associated with PML may have been generated from archetypal JCVs persisting in the patients.
Base Sequence↗
[Association among genotypes of aldehyde dehydrogenase 2 and alcohol consumption].
A large population survey was made for the determination of aldehyde dehydrogenase 2 (ALDH2) genotypes among 630 unrelated Japanese adults by PCR using allele specific primers. The frequencies of three genotypes were 60% ALDH2*1/ALDH2*1 (NN) type, 35% ALDH2*1/ALDH2*2 (ND) type and 5% ALDH2*2/ALDH2*2 (dd) type. Average amounts of alcohol consumption per month for three genotypes were 965.8 g/month for NN type, 629.5 g/month for ND type and 179 g/month for DD type. The differences in average alcohol consumption were significant (p < 0.01) among the three genetic different groups. When male and female date were analyzed separately for these three groups, significant differences among three groups were found in both genders. These results indicated that ALDH2 mutant is an important genetic factor for limiting the alcohol consumption. The frequencies of ALDH2 genotypes were not significantly different between male and female population groups, however, their average alcohol consumption were significantly different between the two groups. Therefore, various social factors and life styles, in addition to genetic factors, might concern in the difference of alcohol consumption.
Adult↗