[A case of Behçet's disease presenting with Hughes-Stovin syndrome (multiple pulmonary arterial aneurysms remitting with corticosteroid therapy)].

A 32-year-old male suffering from thrombophlebitis of the right leg was admitted to our hospital because of massive hemoptysis. Chest radiography showed multiple rounded opacities in the right lung field. Pulmonary angiography revealed multiple aneurysms of the right pulmonary arteries and thromboembolism of the left pulmonary artery. Venography demonstrated obstruction of the deep veins of the right leg and the right femoral vein, and deep vein thrombosis was diagnosed. A more precise diagnosis was Hughes-Stovin syndrome, that is, venous thrombosis especially of the vena cava, accompanied by single or multiple pulmonary arterial aneurysms in young patients. Furthermore, the patient had an aphthous ulcer in the oral cavity, an ulcer in the genital region, leading to a diagnosis of the incomplete type of Behçet's disease. Chest radiographs 3 months after the initiation of corticosteroid showed complete resolution of the aneurysms. Repeated pulmonary angiography also showed partial recanalization of the occluded arteries. This report describes this very rare case of Behçet's disease presenting with Hughes-Stovin syndrome.

Highly diverse TTV population in infants and their mothers.

Infants born to serum HCV-positive 12 mothers were enrolled in the study. Nucleotide sequences amplified by primers deduced from a noncoding region were compared between mothers and their infants. The rates for detection of serum TTV in 12 mothers and their infants were 10/12 (83%) and 9/12 (75%), respectively. Serum TTV DNA was not detected in any infant at 1 month of age, but was detected for the first time between 1.5 and 8 months after birth. Positivity persisted thereafter throughout the follow-up period. In seven randomly selected mother-infant pairs, intrahost TTV heterogeneity was lower in infants than in mothers. Furthermore, one of seven mother-infant pairs showed a high degree of similarity (98.7-100%) in all clones, while in four infants, all nucleotide sequences differed by >10% from those of their mothers. However, the degree of homology in the two mother-infant pairs was 89-98.7% in family 2 and 88.1-99.4% in family 5. In the present study, with only one exception, it was shown that TTV from infants is not identical to TTV from mothers. The mechanism is discussed briefly in this paper.

Amperometric detection of superoxide dismutase at cytochrome c-immobilized electrodes: xanthine oxidase and ascorbate oxidase incorporated biopolymer membrane for in-vivo analysis.

Amperometric measurement of superoxide dismutase (SOD) was carried out at cytochrome c-immobilized monolayers and ascorbate oxidase (AOD)/xanthine oxidase (XOD)/cytochrome c- and (AOD, XOD)/cytochrome c-multilayers. Cytochrome c was covalently immobilized on mercaptopropionic acid-containing self-assembled monolayers on gold. A biopolymer membrane of poly-L-lysine confining XOD and AOD was cast on the monolayer of cytochrome c. While both the cytochrome c-immobilized monolayer and multilayer electrodes show anodic current responses to the generation of superoxide radical, the sensitivity of the multilayer system for the detection of superoxide radical was high relative to that of the monolayer system. In the case of the cytochrome c-multilayer electrodes, the generation of superoxide radical near the sensing element, cytochrome c, resulted in high sensitivity for the detection of superoxide. The use of a XOD and AOD-incorporated poly-L-lysine membrane enabled the detection of the generation of superoxide radical in the presence of L-ascorbic acid. Though L-ascorbic acid could scavenge superoxide radical, the biopolymer membrane confined with AOD will oxidize any L-ascorbic acid that permeated into the membrane. By using the multilayer electrodes, one could measure the activity of SOD in the presence of L-ascorbic acid.

Adsorption properties and activities of lipase on a gold substrate modified by self-assembled monolayers.

The adsorption properties, amount and specific activity of lipase D from Rhizopus delemar were investigated by employing a gold substrate modified with seven kinds of thiol monolayer. Quartz crystal microbalance measurements revealed that the amount of the enzyme adsorbed to the hydrophobic monolayers (e.g. benzenethiol) was much higher than that to the hydrophilic monolayers (e.g. 3-mercaptopropanoic acid). In contrast, lipase D adsorbed to the hydrophilic, 2-amino-1-ethanethiol monolayer showed the highest specific activity, the value being 300-fold higher than for the same enzyme dissolved in an aqueous medium.

Amperometric determination of acetic acid with a trienzyme/poly(dimethylsiloxane)-bilayer-based sensor.

A trienzyme sensor for the amperometric determination of acetic acid was prepared by immobilizing acetate kinase (AK), pyruvate kinase (PK), and pyruvate oxidase (PyOx) on a poly(dimethylsiloxane) (PDMS)-coated electrode. AK catalyzes the phospho-transferring reaction between acetic acid and ATP to form ADP; PK, the phospho-transferring reaction between ADP and phosphoenolpyruvate to form pyruvic acid; and PyOx, the oxidation of pyruvic acid with oxygen. The oxygen consumption could be monitored by using the PDMS-coated electrode without interference from the PyOx reaction product, hydrogen peroxide. Thus, the concentration of acetic acid (5 microM-0.5 mM) could be determined from the decrease in the cathodic current at -0.4 V vs Ag/AgCl. This is the first example of a biosensor that can be used for the determination of acetic acid in ethanol-containing food samples. The acetate-sensing electrode could be used for more than one month.

Detection rates of TT virus DNA in serum of umbilical cord blood, breast milk and saliva.

To date, the routes of mother-to-infant transmission of TT virus (TTV) have not been fully elucidated. The present study examines the detection rates of TTV DNA in the serum of pregnant Japanese women and in cord blood at the time of delivery, as well as in the saliva and breast milk of mothers one-month postpartum. Primers derived from the well-known translated region N22 (N22 system), as well as the untranslated region (UTR system) were used. The prevalence of TTV DNA in the serum of pregnant women was found to be 11.9% (19/160) using the N22 system and 72.4% (55/76) using the UTR system. No TTV DNA was detected in the cord blood samples (0/160) when the N22 system was used for detection but TTV DNA was detected in 11.8% (7/76) of samples studied with the UTR system. Using the N22 system, TTV DNA was not detected in breast milk, but was detected in saliva. However using the UTR system, TTV DNA was detected in both specimens. These results imply that some babies are vertically infected with TTV via cord blood at the time of delivery or via breast milk or saliva. However, further research is necessary to confirm this hypothesis. polymerase chain reaction; pregnant women; horizontal route of transmission

TT virus infection in Japanese children: isolates from genotype 1 are overrepresented in patients with hepatic dysfunction of unknown etiology.

The pathogenecity of the TT virus (TTV) especially during childhood remains obscure. We investigated the prevalence of TTV in 40 patients with non-A to C hepatic dysfunction (non-A to C hepatic dysfunction group). Five patients with fulminant hepatitis of unknown etiology were enrolled in this group. We also examined 380 children without a history of transfusion or liver disease (control group). Subsequently, the genotypes of TTV strains isolated were analyzed in terms of their nucleotide sequences including 222 bp in the open reading frame 1 region. The prevalence of serum TTV DNA was 10/40 (25%) in the non-A to C hepatic dysfunction group and 25/380 (7%) in the control group. Sixty-six percent (23/35) of all examined cases exhibited either genotype 1 or 2. However, assessment of genotype in the non-A to C hepatic dysfunction group (10 cases) revealed a higher prevalence of genotype 1 than of all other genotypes (80% vs. 20%). This result differed significantly from that of the control group (25 cases; 32% vs. 68%). Such overrepresentation of genotype 1 suggests that this type of TTV strain is associated with the development of hepatic dysfunction of unknown etiology in Japanese children.

The prevalence of serum GB virus C/hepatitis G virus RNA and anti-E2 in Japanese children without a history of blood transfusion.

The prevalence of serum GB virus C (GBV-C)/Hepatitis G virus (HGV) RNA and anti-E2 was investigated in Japanese children younger than 16 years of age without a history of blood transfusion and the family members of serum GBV-C/HGV RNA-positive children. The prevalences of serum GBV-C/HGV RNA and anti-E2 were 0.5% (5/1000) and 0% (0/330), respectively. Viral RNA was also detected in the mothers of all five GBV-C/HGV RNA-positive children and in two of their siblings. Sequence determinations indicated the likelihood of mother-to-infant transmission in all cases. The presence of the virus persisted for at least 10-18 months in all 5 children, without any appearance of anti-E2.

Prevalence of TTV DNA among children with a history of transfusion or liver disease.

The prevalence rates of serum TT virus (TTV) DNA among children with or without a history of transfusion or liver disease were studied by polymerase chain reaction (PCR) using either the Okamoto primer set or the Takahashi primer set developed more recently. Using Okamoto and Takahashi primer sets, the prevalence rates were 31.6% (12/38) and 78.9% (30/38), respectively, for children with a history of blood transfusion (including malignant and non-malignant groups) and 6.7% (2/30) and 60% (18/30), respectively, for children without a history of blood transfusion. Among pregnant women, these rates were 12.9% (4/31) and 61.3% (19/31), respectively. On the other hand, the prevalence rates were 0% (0/16) and 50% (8/16), respectively, in hepatitis B patients, 21.4% (3/14) and 71.4% (10/14), respectively, for hepatitis C patients, and 20.0% (9/45) and 57.8% (26/45), respectively, for non-A to C hepatitis patients (including 27 acute hepatitis patients, 5 fulminant patients and 13 chronic hepatitis patients). In this study, the prevalence rates determined by the Takahashi primer set tended to be 2-9 times higher than those determined using the Okamoto primer set. These results suggest that TTV infection is widespread among Japanese children. Furthermore, blood transfusion does not appear to be the major route of infection. The similar prevalence rates between control children and children with various types of hepatitis using the Takahashi primer system suggest that TTV infection does not play a direct causative role in the development of liver disease in children.

[A case of pulmonary tuberculosis with acute renal failure caused by readministration of rifampicin].

We report a case of pulmonary tuberculosis with acute renal failure caused by readministration of Rifampicin (RFP). A 73 year-old man was admitted to a certain hospital complaining with dyspnea on exertion. As his sputum smear was positive for acid-fast bacilli, he was transferred to our hospital for the isolation and treatment. He was diagnosed as lung tuberculosis and was administrated RFP, Isoniazid (INH) and Ethambutol (EB). On the 20th day after the initiation of treatment, the administration of drugs were suspended, because of liver dysfunction. After recovery of liver dysfunction, we have readministered antituberculous drugs, starting with EB, then INH, and finally RFP. On the 22nd day after the readministration of RFP, acute renal failure was observed. All medications were suspended and we started treatment with hydration and furosemide. His renal function recovered after 7 weeks. Histopathological examination of the kidney revealed interstitial infiltration and tubular nephritis. According to the histopathological examination and the clinical course, we concluded acute renal failure was induced by the readministration of RFP. This case suggests that we have to pay attention to renal side effect of RFP in the course of readministration.

[Outbreak of multidrug-resistant Mycobacterium tuberculosis infection in the middle and advanced age].

We experienced a small outbreak of multidrug-resistant Mycobacterium tuberculosis infection (MDR-TB) among persons of in the middle and advanced age. The index case was 48-year-old man, and had complained productive cough since January 1996. He visited a doctor due to his symptom, and chest X-ray revealed cavitary lesion and sputum smear was positive for tubercle bacilli. He could not continue his admission because of his absence without leave and drinking, he was discharged on Day 54. The drug resistance was observed for INH (0.1 microgram/ml), RFP, and SM. Later, case 2, 52-year-old male, and case 3, 43-year-old-male, who were companions in mah-jongg with the index case, were diagnosed as pulmonary tuberculosis. The analysis of restriction fragment length polymorphism (RFLP) was done on 3 strains, and all showed the same pattern. Among other companions in mah-jongg with the index case, case 4, 28-year-old male, was treated as MDR-TB, and the drug resistance pattern was the same to that of the index case, but the details were unknown. Case 5, 65-year-old male, was diagnosed as drug sensitive pulmonary tuberculosis, thus he might incidentally suffer from pulmonary tuberculosis at the same time. Case 6, 46-year-old male, who had been treated for alcoholic liver cirrhosis, was introduced to our hospital as his sputum smear was positive, and the drug resistance pattern was observed similar to that of the index case. All the companions in mah-jongg suffered from MDR-TB except case 5. The RFLP analysis showed that the index case, case 2, and case 3 were caused by the same strain of M. tuberculosis. The drug resistance pattern of, case 4 and case 6 was the same to that of the index case. Based on these findings, it is highly suspected that this small outbreak was originated from the index case.

Route of TT virus infection in children.

TT virus (TTV) is a novel viral agent, detected recently in non-A to E hepatitis cases. Little is known about its natural history or routes of transmission in childhood. For the detection of serum TTV DNA, semi-nested polymerase chain reaction (PCR) was carried out using TTV-specific primers and TTV nucleotide sequences were determined by the dideoxy chain-mediated termination method. Five of the 70 children studied (including 20 hepatitis B virus [HBV] carriers, 40 children born to HBV carrier mothers and 10 children born to hepatitis C virus [HCV] carrier mothers) had serum TTV DNA. Three of the 5 children had siblings (4 in total), so that a total of 9 children were studied to determine the time of initial serum TTV DNA detection. In the 8 seropositive children, the time of serum TTV DNA detection ranged from 6 to 14 months after birth, and TTV DNA persisted thereafter throughout the follow-up period. The TTV DNA-negative child was assessed most recently at 6 months of age. TTV DNA was detected in only 2 of the 4 mothers tested (families 2 and 3). When 271-bp TTV DNA fragments from each of the 8 children were sequenced, the degree of homology between siblings in families 1-3 was 100%, 99.5%, and 92.3%, respectively. The degree of homology between child-mother pairs of families 2 and 3 was 99.5-100% and 62. 6-63.9%, respectively. The distribution of different TTV strains was consistent within families, except for family 3. None of the TTV-infected children had elevated levels of alanine aminotransferase or clinical signs of liver disease.

Evaluation of a new anti-HCV-assay kit for anti-HCV screening in early childhood.

Sera of 20 children falsely identified as positive for hepatitis C virus antibody (Anti-HCV) by a second generation anti-HCV-assay kit (Imucheck-HCV Ab "Kokusai") were re-tested using a new third generation anti-HCV-assay kit (Imucheck x F-HCV C50 Ab "Kokusai"). Seventeen of the samples were reclassified as negative and only three remained positive. Changing well solids in the anti-HCV-assay kit from casein to bovine serum albumin appears to have improved the false-positive rate, most likely as a result of decreased non-specific adsorption of casein antibodies.

[Tuberculosis microepidemic in a commuter bus].

A tuberculosis microepidemic in a commuter bus was reported. Index patient was a 22-year-old woman who was an employee of an electronic company. An abnormal shadow was found on her chest roentgenogram during an annual medical check-up in June, 1996. As her sputum smear was Gaffky 6, she was admitted to our hospital for medication. Extraordinary examinations including PPD skin test and chest X-ray were carried out on 49 employees of the company in October, 1996. As the result of these examinations, the distribution of maximum diameters of erythema in PPD skin test showed bimodal distribution, and tuberculosis was discovered in two patients by Chest X-ray examination. Moreover, preventive administration of Isonicotinic acid hydrazide (INH) was indicated for 3 employees based on very strong skin reaction to PPD. These five employees were working separately from the index patient and had little contact with the patient in the work places, but using a same commuter bus. Therefore, we strongly suspect that they were infected from the index patient not in the work place but in the commuter bus. The air-conditioning of the bus used a closed recirculation system, hence insufficient ventilation in the bus contributed to the spread of tuberculosis infection.

[A case of pulmonary tuberculosis case with pancytopenia accompanied to bone marrow gelatinous transformation].

A 45-year-old man did not visit a doctor in spite of his complains, cough and sputum lasting, for six months, and he finally could not eat without beer, and as a result, he lost his body weight and currently 52 kg. He became unconsciousness, was carried to a hospital, and was referred to our hospital. His sputum examination for acid fast bacilli was smear positive, Gaffky 6, for M. tuberculosis. His chest roentogenogram revealed large cavitary lesions in bilateral lung fields. On blood examination, WBC was 1100/microL, RBC was 256 x 10(4)/microL, and PLT was 13.4 x 10(4)/microL. Total protein was 4.7 g/dl, albumin was 1.9 mg/dl, and total cholesterol was 65 mg/dl. We tried to aspirate bone marrow from his sternum, but it was impossible. Hence we did biopsy of his ilium. The pathology of his bone marrow revealed gelatinous transformation. It was thought that the marked delay in visiting a doctor caused general consumption and loss of apetite, thus led to gelatinous transformation and finally pancytopenia.

Detection rates of TT virus among children who visited a general hospital in Japan.

Recently, genomic DNA of the novel TT virus (TTV) was isolated from patients suffering from posttransfusion hepatitis of unknown etiology. We examined sera from 197 children who visited the Department of Pediatrics at Toyohashi National Hospital. Sera were tested for TTV DNA by seminested polymerase chain reaction (PCR) using a set of primers synthesized according to the published TTV sequence. Ten children were found to be positive for TTV (5.1%). All positive PCR products were directly sequenced in both directions using a fluorescent dye terminator cycle sequencing system. The sequences were compared by a multiple sequence alignment and a phylogenetic tree was constructed. The phylogenetic tree showed that two of the TTV isolates found in the present experiment did not belong to any of the phylogenetic groups previously reported.

Prevalence of GB virus C/hepatitis G virus ribonucleic acid and anti-hepatitis G virus-E2 antibodies among Japanese children with histories of transfusions or with liver diseases.

To clarify the prevalence of Japanese children thought to be at a risk for infection with GB virus-C (GBV-C)/hepatitis G virus (HGV), we investigated the detection rates of serum GBV-C/ HGV ribonucleic acid (RNA) by reverse transcription-seminested PCR and serum anti-HGV-E2 antibody by ELISA in 162 children with histories of blood or plasma product transfusions or with liver diseases and performed phylogenetic analysis of the 5' noncoding region sequences of GBV-C/HGV genomes. Children with histories of transfusions were divided into those who had been treated with antineoplastic agents for malignant diseases (malignant group) and those who had received transfusions for nonmalignant diseases (nonmalignant group). Children with liver diseases were divided into hepatitis B (HBV), hepatitis C (HCV), and non-A-C hepatitis groups. We detected GBV-C/ HGV RNA in 11 of 33 (33.3%) and anti-HGV-E2 in 1 of 27 (3.7%) children in the malignant group and in 3 of 56 (5.4%) and 1 of 53 (1.9%) children, respectively, in the nonmalignant group. Neither GBV-C/HGV RNA nor anti-HGV-E2 was detected in the HBV and non-A-C hepatitis groups. GBV-C/HGV RNA and anti-HGV-E2 were detected in 7 of 23 (30.4%) and in 1 of 18 (5.6%) children, respectively, in the HCV group. All children positive for either GBV-C/HGV RNA or anti-HGV-E2, except one whose route of GBV-C/HGV infection suggested mother-to-infant transmission, had histories of transfusions. The phylogenetic analysis showed that all isolates in this study were divisible into three groups and that most of them were clustered into group 3 (Asian group).

High-throughput flow-injection analysis of glucose and glutamate in food and biological samples by using enzyme/polyion complex-bilayer membrane-based electrodes as the detectors.

The concentration of glucose was determined by a combination of flow injection analysis (FIA) with amperometric enzyme sensor detection. The enzyme sensor was prepared by immobilizing glucose oxidase on an electrode coated with a polyion complex layer consisting of poly-L-lysine and poly(4-styrenesulfonate). The inner, polyion complex layer was useful for preventing electrochemical interferents (e.g., L-ascorbic acid, uric acid and acetaminophen) from reaching the electrode surface, which was effective for reducing the interferential responses upon the injections of biological and food samples. The sensor-based system could be used for the determination of glucose from 10 microM to 3 mM with the sampling rate of 180 h-1, and was stable for more than 2 months. An FIA system for determining L-glutamic acid (3 microM-0.5 mM) was also prepared by using an enzyme electrode based on a glutamate oxidase/polyion complex-bilayer as the detector.

[A case of cutaneous tuberculosis associated with steroid therapy for mixed connective tissue disease].

Patients receiving immunosuppressive therapy, such as adrenocorticosteroids, are high risk groups of tuberculosis. We report a case of cutaneous tuberculosis associated with steroid therapy for mixed connective tissue disease. A 63-year-old female was hospitalized after 6 months' treatment with prednisolone for connective tissue disease and bilateral abnormal shadows were revealed on her chest X-ray films. As her sputum smear was positive for acid-fast bacilli, the patient was transferred to our hospital for isolation and treatment. After three months' treatment with INH, RFP and EB, she complained the swelling of her left palm, left arm, and right leg, and skin puncture was performed. As smears of fluid aspirated from the swelling showed acid-fast bacilli, and fluid PCR tests showed positive for M. tuberculosis, she was diagnosed as cutaneous tuberculosis (scrofuloderma). In spite of administration of antituberculous agents, the swelling showed little improvement. Therefore, the dose of prednisolone was reduced and cutaneous lesions were resected by surgery. High risk of tuberculosis should be considered when a patient administered immunosuppressive drugs, such as adrenocorticosteroids.