An outbreak of psittacosis in a bird park in Japan.

An outbreak of psittacosis related to a bird park occurred in Matsue City, Shimane Prefecture, Japan, during winter 2001. Seventeen cases of psittacosis (12 visitors, three staff, and two student interns) were confirmed. A cohort study was conducted among the park staff and students to determine the risk factors for the development of acute serologically confirmed psittacosis (SCP) infection. Being 'bird staff' had an increased risk of SCP infection (RR 3.96, 95% CI 1.48-10.58). Entering the staff building, where ill birds were maintained without proper isolation, was also associated with an increased risk of SCP infection (RR 3.61, 95% CI 1.03-12.6). Isolation of ill birds and quarantine measures were found to be insufficient. Dehumidifiers and a high-pressure water spray under a closed ventilation environment may have raised the concentration of Chlamydophila psittaci in the hothouses. Bird park staff and visitors should be educated about psittacosis.

Human babesiosis in Japan: epizootiologic survey of rodent reservoir and isolation of new type of Babesia microti-like parasite.

We have carried out epizootiologic surveys at various sites in Japan to investigate wild animals that serve as reservoirs for the agents of human babesiosis in the country. Small mammals comprising six species, Apodemus speciosus, Apodemus argenteus, Clethrionomys rufocanus, Eothenomys smithii, Crocidura dsinezumi, and Sorex unguiculatus, were trapped at various places, including Hokkaido, Chiba, Shiga, Hyogo, Shimane, and Tokushima Prefectures. Animals harboring Babesia microti-like parasites were detected in all six prefectures. Inoculation of their blood samples into hamsters gave rise to a total of 20 parasite isolates; 19 were from A. speciosus, and the other 1 was from C. rufocanus. Sequencing of the parasite small-subunit rRNA gene (rDNA) sequence revealed that 2 of the 20 isolates were classified as Kobe type because their rDNAs were identical to that of the Kobe strain (the strain from the Japanese index case). The other 18 isolates were classified as a new type, designated the Hobetsu type, because they all shared an identical rDNA sequence which differed significantly from both that of Kobe-type isolates and that of northeastern United States B. microti (U.S. type). The parasites with Kobe-, Hobetsu- and U.S.-type rDNAs were phylogenetically closely related to each other but clearly different from each other antigenically. The isolates from rodents were demonstrated to be infective for human erythrocytes by inoculation into SCID mice whose erythrocytes had been replaced with human erythrocytes. The results suggest that a new type of B. microti-like parasite, namely, the Hobetsu type, is the major one which is prevalent among Japanese wild rodents, that A. speciosus serves as a major reservoir for both Kobe- and Hobetsu-type B. microti-like parasites, and that C. rufocanus may also be an additional reservoir on Hokkaido Island.

Isolation, characterization and mutation analysis of PEX13-defective Chinese hamster ovary cell mutants.

We isolated peroxisome biogenesis mutants ZP128 and ZP150 from rat PEX2 -transformed Chinese hamster ovary (CHO) cells, by the 9-(1'-pyrene)nonanol/ultraviolet method. The mutants lacked morphologically recognizable peroxisomes and showed a typical peroxisome assembly-defective phenotype such as a high sensitivity to 12-(1'-pyrene)dodecanoic acid/UV treatment. By means of PEX cDNA transfection and cell fusion, ZP128 and ZP150 were found to belong to a recently identified complementation group H. Expression of human PEX13 cDNA restored peroxisome assembly in ZP128 and ZP150. CHO cell PEX13 was isolated; its deduced sequence comprises 405 amino acids with 93% identity to human Pex13p. Mutation in PEX13 of mutant ZP150 was determined by RT-PCR: G to A transition resulted in one amino acid substitution, Ser319Asn, in one allele and truncation of a 42 amino acid sequence from Asp265 to Lys306 in another allele. Therefore, ZP128 and ZP150 are CHO cell lines with a phenotype of impaired PEX13.

Newly identified Chinese hamster ovary cell mutants defective in peroxisome assembly represent complementation group A of human peroxisome biogenesis disorders and one novel group in mammals.

We isolated peroxisome biogenesis-defective mutants from rat PEX2-transformed Chinese hamster ovary (CHO) cells, using the 9-(1'-pyrene)nonanol/ultraviolet method. A total of 18 mutant cell clones showing cytosolic localization of catalase were isolated. By complementation group (CG) analysis by means of PEX cDNA transfection and cell fusion, cell mutants, ZP124 and ZP126, were found to belong to two novel CGs of CHO mutants. Mutants, ZP135 and ZP167, were also classified to the same CG as ZP124. Further cell fusion analysis using 12 CGs fibroblasts from patients with peroxisome deficiency disorders such as Zellweger syndrome revealed that ZP124 belonged to human CG-A, the same group as CG-VIII in the United States. ZP126 could not be classified to any of human and CHO CGs. These mutants also showed typical peroxisome assembly-defective phenotypes such as severe loss of catalase latency and impaired biogenesis of peroxisomal enzymes. Collectively, ZP124 represents CG-A, and ZP126 is in a newly identified CG distinct from the 14 mammalian CGs previously characterized.

Survey of human group C rotaviruses in Japan during the winter of 1992 to 1993.

Fecal specimens from patients with acute diarrhea were collected from 10 prefectures in Japan over a 6-month period (November 1992 to April 1993), and the specimens that were negative for human group A rotaviruses were screened for the presence of human group C rotaviruses (CHRVs) by the reverse passive hemagglutination test. Of 784 specimens examined, 53 samples (6.8%) that were collected in 7 of 10 prefectures were positive for CHRV, indicating that CHRVs are widely distributed across Japan. Most of the CHRV isolates were detected in March and April, and CHRVs mainly prevailed in children ages 3 to 8 years. The genome electropherotypes of eight strains isolated in five individual prefectures were surprisingly similar to each other and were different from those of CHRV strains isolated to date. The outer capsid glycoprotein (VP7) gene homologies of the isolates retrieved in 1993 were subsequently analyzed by the dot blot hybridization method. As a result, the VP7 genes of the isolates revealed very high levels of homology not only with each other but also with the VP7 gene of the OK118 strain isolated in 1988. These results suggest that a large-scale outbreak of CHRV occurred during the winter of 1992 and 1993 in Japan.

Comparison of gelatin particle agglutination and hemagglutination inhibition tests for measles seroepidemiology studies.

The prevalence of measles antibody in Japan was surveyed with a newly developed gelatin particle agglutination (PA) test, and the results compared with those of the hemagglutination inhibition (HI) test. The two age-distribution curves of the PA antibody-positive rates at > or = 1:8 and > or = 1:32 were almost the same in all the age groups, except the less-than-1-year-old group for which the rate at > or = 1:8 was higher than that at > or = 1:32 (p < 0.05, chi 2 test). In the vaccinated children, all groups older-than-1-year of age had antibody-positive levels of 96% or more. In contrast, in the unvaccinated children, there was a sharp increase in antibody-positive rates between the 1- and 4-year-old groups, indicative that about 80% of the children were infected by wild measles virus at these ages. A significant number of PA antibody-positive specimens were antibody-negative (< 1:8) by HI. The percentage of specimens in this category, PA (+) but HI (-), was greatest in infants less than one year old, and least in young children, but it increased with age to 97% of the HI (-) specimens from adults of more than 20 years of age. The PA test therefore detected some measles antibodies that HI could not. This test is simple and useful for making serosurveys in both developed and developing countries.

Determination of l-phenylalanine in serum by flow-injection analysis using immobilized phenylalanine dehydrogenase and fluorimetric detection.

A flow-injection system with an immobilized enzyme reactor is proposed for the determination of l-phenylalanine. Phenylalanine dehydrogenase from Rhodoccus sp. M4 was immobilized on tresylated poly (vinyl alcohol) beads (13 mum) and packed into a stainless-steel column (5 cm x 4 mm i.d.). Serum sample was deproteinized with tungstic acid and filtered through an ultrafiltration membrane. The sample solution (30 mul) was injected into the carrier stream (water). The NADH formed was detected at 465 nm (excitation at 340 nm). The calibration graph was linear for 0.9-600 mum l-phenylalanine; the detection limit was 0.3 mum. The sample throughout was 25 h(-1) without carryover. The half-life period of the immobilized enzyme was 23 days.

[Serosurvey for spotted fever group rickettsial infection in vertebrates in Shimane Prefecture].

We carried out a survey for the prevalence of antibodies to the spotted fever group (SFG) rickettsia in vertebrates such as dogs, cattle, deer, and mice in Shimane Prefecture. Rickettsia japonica was employed as antigen in indirect immunofluorescence (IF) tests. The experiment for natural infections using decoy animals was performed in the field of the endemic area. 1. Among 115 street dogs, 18.3% possessed the antibodies against SFG rickettsia, while all of the 8 hunting dogs had the antibodies. 2. Among 234 cattle tested, 17.9% possessed the antibodies. IF titers were 1:40 to 1:160 (mean 1:68). 3. Among 69 wild deer, 92.7% possessed the antibodies ranging between 1:40 and 1:640, which showed the highest IF titers (mean 1:89) among those of the examined vertebrates. 4. The incidence of the antibodies in Apodemus speciosus, Apodemus argenteus and Eothenomys smithi smithi mice were 16.5, 4.3 and 0%, respectively. The incidence of the antibodies against SFG rickettsia in mice captured in the endemic area was significantly higher (22.8%) than that in non-endemicarea (10.4%). Difference in the incidence of antibody-positive mice was also observed within the endemic area. Therefore, we concluded that the infection of mice was restricted to a limited area. 5. No significant rise in IF titers was observed in decoy animals that had been infested with ticks in the endemic area.

[Do blood glucose levels in the elderly have different metabolic significance than in non-elderly subjects?].

Diagnostic criteria of diabetes mellitus in elderly people are still controversial. The major questions are whether (1) effects of hyperglycemia in generating diabetic complications are the same as in younger diabetics, (2) too many diabetics might be diagnosed in the elderly by the criteria proposed by WHO and Japan Diabetes Society. To answer these questions, we carried out two studies. Blood glucose data were collected from population-based studies and from voluntary health check-up studies. Fasting blood glucose values were classified by sex and the age (-49, 50-64, 65-years old). Population-based studies showed no age-dependent change in blood glucose. However, data from health check-up studies showed an increase in median values from the group aged 50-64 and the 65 and older group. Two-hour values after glucose tolerance tests were classified by age and the fasting blood glucose (-99, 100-119, 120-139, and 140- mg/dl). The eldest and 140- mg/dl group showed the highest 2-hour values. Effects of hyperglycemia on the appearance of diabetic complications were studied in patients being follow-up in three diabetes clinics. Patients who has a history of one year or less and without any retinopathy on the first visit to the clinic were registered. Data were analyzed by the Kaplan-Meier method and the appearance of retinopathy was used as the end point. There were no difference in the rate of appearance of retinopathy between the age groups (-49, 50-64 and 65-years old). No age effect on the hyperglycemia-dependent appearance of retinopathy could be observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Rapid DNA diagnosis of herpes simplex virus serotypes.

The presence of nucleotide sequences specific for each of herpes simplex virus (HSV) serotypes was demonstrated. These sequences were applied for dot DNA-DNA hybridization and for PCR for rapid DNA diagnosis of HSV infections. These sequences were found by molecular cloning of HSV-DNA fragments after digestion of DNA by KpnI enzyme. The type 1-specific sequence was found around the 5' end of BamHI B-fragment in the L region of type 1 DNA (corresponds to alpha gene 27, promoter-regulatory region) and the type 2-specific sequence was around the junction region of the L and S of type 2 DNA (corresponds to a' sequence). Both simple dot blot hybridization and PCR of HSV DNA's, employing these type-specific nucleotide sequences, were proven to be much more useful than immunofluorescence in terms of type-specific diagnosis of HSV infections.

[A case report of spotted fever group rickettsiosis first encountered in Shimane Prefecture, Japan].

Rickettsial infections of a spotted fever group have recently been experience in Japan. Although there have been 50 cases reported by this time, they were exclusively distributed in the areas along the Pacific coast. We report in this paper the first case of spotted fever group rickettsiosis in Shimane prefecture, an area faced to the Japan Sea. A 57 year-old man with high fever, general fatigue and rash was admitted to the hospital of Shimane Medical University at the end of September 1987, who had been treated with cefaclor for a few days without effect before his visit to the University Hospital. Physical examination revealed erythematous eruption, eschar on the left side of the abdomen, and generalized lymphadenopathy. Hepatosplenomegaly was not detected. Laboratory studies showed normal leukocyte counts with relative lymphopenia, high ESR and increased CRP. Transaminase levels were slightly elevated. Paul-Bunnel and Weil-Felix tests resulted in negative. Specific immunofluorescence tests demonstrated that IgG and IgM antibodies of acute-phase serum to Rickettsia japonica, a spotted fever group rickettsia isolated from patients in Japan, were not detected, while both IgG and IgM antibody titers of convalescent-phase serum increased to 1:320. Convalescent-phase serum reacted at significantly lower titers with R.typhi as well as other pathogenic spotted fever group rickettsiae. The patient was treated with ofloxacin for six days and then with combination of ofloxacin and minocycline. It should be noted that clinical symptoms such as high fever and general fatigue disappeared during a single therapy with ofloxacin.(ABSTRACT TRUNCATED AT 250 WORDS)

A change of urinary proteins from a glomerular pattern to a tubular pattern during a late diuretic phase of acute renal failure.

The proteinuria on admission, on the 3rd day of the disease, of a 15-year-old male patient with acute renal failure (vasomotor nephropathy) showed a glomerular proteinuria-predominant pattern, and had a normal activity of N-acetyl-beta-D-glucosaminidase, indicating little proximal tubular damage. During a late diuretic phase, the excretion of N-acetyl-beta-D-glucosaminidase in the urine increased in accordance with increases of sodium excretion and urine flow, and then decreased to the normal range on the 20th day of the disease. Urine during the late diuretic phase showed a tubular proteinuria pattern and finally returned to a normal pattern on the 47th day of the disease. Thus, transient proximal tubular damage was evidenced in the late diuretic phase of the patient.

Isolation and characterization of a cold-sensitive strain of coxsackievirus A10.

A coxsackievirus A10 strain, isolated from a clinical specimen from a patient with pharyngitis, was characterized with respect to its growth properties in different cultured cells and at different incubation temperatures. This virus multiplied within cultured cells and produced cytopathogenic effects, whereas a prototype strain of coxsackievirus A10 did not. The isolate multiplied efficiently in cultured cells at 37 degrees C but its replication was markedly restricted at 32 degrees C. Temperature shift experiments indicated that the cold-sensitive event affected the late function(s) of the virus.

Characteristic regional prevalence of HTLV-I in Shimane Prefecture.

Serological survey for human T-lymphotropic virus type I (HTLV-I) was performed in Shimane Prefecture, Japan, located on the coast of Japan sea. The rates of anti-adult T-cell leukemia-associated antigen-positive sera were 4.2% and 6.9% respectively for males and females. The rate increased with age, particularly in females. From the geographical point of view, the Oki Islands and a very small focus in Shimane-cho had high rates of anti-adult T-cell leukemia-associated antigen positives. Even within the Oki Islands, there was a big difference in the rate of seropositives between Dozen and Dogo which are only 10 km apart. The focus, Shimane-cho, has a population of 299. Even within a small area, there is a tendency of HTLV-I carriers to form a very narrow focus.