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Biomedical subjects

H Kuroki

Publications and source records attributed to H Kuroki.

At least 37 records · Page 2Linked to original sources

[The incidences of virulence genes in Escherichia coli strains from sporadic diarrheal children].

To investigate the incidence of diarrheagenic Escherichia coli among E. coli strains screened by commercially available O-antigen antisera, we used PCR to isolate 8 virulence genes (eae, bfpA, IpaH, LT, ST, VT1, VT2, and aggR) in 184 E. coli strains sampled from sporadic diarrheal children in our district. eae and bfpA are the localized adherence factor genes of enteropathogenic E. coli (EPEC). IpaH is the invasion antigen gene of enteroinvasive E. coli (EIEC), LT and ST are the toxin genes of enterotoxigenic E. coli (ETEC), VT1 and VT2 are the toxin genes of enterohemorrhagic E. coli (EHEC), and aggR is the adherence factor gene of enteroaggregative E. coli (EAggEC). The results were as follows: eae, 7 (3.8%); bfpA, 0 (0%); IpaH, 0 (0%); LT, 0 (0%); ST, 2 (1.1%); VT, 5 (2.7%); aggR, 8 (4.3%). Seven isolates with eae did not have bfpA, and did not show a localized adherence to HeLa cells. Seven of the 8 isolates with aggR showed aggregative adherence to HeLa cells, while their O-serotypes of them were O111:H21 or O111:HUT. Because of the low incidence of the virulence gene, the commercially available O-antigen antisera was not expected to be useful for the screening of diarrheagenic E. coli, except for EHEC and EAggEC. EAggEC may be important as a pathogen of sporadic diarrhea of children as well as EHEC.

Child↗

[Clinical analysis of patients with bacterial meningitis in childhood and reevaluation of rapid antigen detection methods].

Twenty-eight cases of bacterial meningitis during the recent ten years were analyzed retrospectively, and the following results were obtained. 1. Pathogens were as follows; H. influenzae 13 (46.4%), S. pneumoniae 8 (28.6%), S. agalactiae 4 (14.3%), E. coli 2 (7.1%), and L. monocytogenes 1 case (3.6%). 2. Twelve out of the thirteen H. influenzae cases were caused by serotype b (Hib), and 2 strains were beta-lactamase producer. Fifty percent of the S. pneumoniae cases were caused by penicillin-resistant strains. And all these resistant strains belonged to serotype 19 or 23. 3. Underlying diseases related to the onset of meningitis were found in 46% of the cases, and these consisted of CNS shunt operated 5, asplenia or polysplenia 2, Mondini's anomaly 1, sacral dermal sinus 1, and neonate 4 cases. 4. Prognosis of these cases were three deaths, four with neurologic sequelae, and twenty-one complete recoveries. 5. On admission, 85% (17/20) of the cases were diagnosed correctly by the rapid antigen detection. Sensitivity and specificity of the rapid antigen detection by using latex particle agglutination is 90% and 100% in the Hib cases, and 83% and 100% in the S. pneumoniae cases respectively. Moreover, the bacteriologically unknown 2 cases caused by parenteral partial treatment were also diagnosed by the detection of antigen in concentrated urine.

Antigens, Bacterial↗

Molecular epidemiological study of Haemophilus influenzae serotype b strains obtained from children with meningitis in Japan.

We report an epidemiological study of 30 Haemophilus influenzae serotype b (Hib) strains derived from the cerebrospinal fluid of children with meningitis. The Hib strains were biotyped, tested for beta-lactamase production, and genotyped by long PCR-ribotyping, random amplified polymorphic DNA (RAPD) analysis, and genomic DNA restriction fragment length polymorphism (RFLP) analysis by pulsed-field gel electrophoresis (PFGE). The phenotypic study characterized 22 of the strains (73%) as biotype I. A genotypic study using long PCR-ribotyping with HaeIII restriction digestion showed no polymorphisms among these 30 Hib strains, but RAPD analysis with two sets of primers demonstrated two distinctive subtypes: one typical of the strains of biotype group II and the second characteristic of the strains of biotype groups I and IV. Each RAPD group was subtyped into several genotypic groups by PFGE-RFLP with SmaI digestion. The genotyping of clinically isolated Hib strains may help to elucidate transmission routes in community infections, endemicity, and the reasons for vaccine failure.

Child, Preschool↗

Segmental square spinal instrumentation for posterior lumbar spinal fixation.

The purpose of this study is to evaluate the clinical results of operations using a new spinal instrumentation for posterior fixation, called segmental square spinal (3-S) instrumentation. The 3-S instrument consists of two pairs of hooks to clasp the interarticular portion of the lamina bilaterally, two horizontal bars to connect the hooks on the right and left, and rods to connect the hooks side by side on the top and bottom. Twenty-seven patients who had degenerative lumbar disorders underwent operations with the 3-S instrument. Rigid fixation was obtained in all cases immediately after the operation. Bone union rates were 91% (21/23) with posterolateral fusion. The 3-S instrument seems to be suitable for spinal disorders in which anterior spinal fusion is not necessary, especially for degenerative spinal disorders.

Adult↗

[Comparative study on metabolism of three tetrachlorobiphenyls with animal liver microsomes].

In vitro metabolism of 3,5,3',4'-, 3,5,3',5'- and 2,4,3',4'-tetrachlorobiphenyls (TCBs) was studied using liver microsomes from rats, guinea pigs and hamsters. 3,5,3',4'-TCB was metabolized to 4-hydroxy-3,5,3',4'-TCB with liver microsomes of 3-methyl-cholanthrene (MC)- and 3,4,5,3',4'-pentachlorobiphenyl (PenCB)-treated rats but not of phenobarbital (PB)-treated ones. This result suggests that a MC-inducible cytochrome P450 isoform, probably CYP1A1, is more important in the in vitro metabolism of 3,5,3',4'-TCB in rat liver and that the isoform attacks the 3,5-dichloro-substituted phenyl ring more predominantly than 3,4-dichloro-substituted one. In 3,5,3',5'-TCB metabolism, liver microsomes from MC- and 3,4,5,3',4'-PenCB-treated hamsters formed 4-hydroxy-3,5,3',5'-TCB to a similar extent to rats reported previously. Guinea pig liver microsomes formed no metabolite. In 2,4,3',4'-TCB metabolism, PB accelerated 3-, 5- and 4-hydroxylations in guinea pigs and also 3- and 5-hydroxylations in hamsters, suggesting the involvement of a PB-inducible P450 isoform, presumably P450GP-1 and P450HPB-1, respectively. On the other hands, MC- and 3,4,5,3',4'-PenCB-treatment resulted in the marked increase of 4-hydroxylation in hamsters, but in the suppression of 4-hydroxylation in guinea pigs. From these results, it is suggested that the hydroxylation of coplanar TCBs such as 3,5,3',4'- and 3,5,3',5'-TCB is catalyzed by a MC-inducible P450 in rats and hamsters, whereas non-coplanar TCBs such as 2,4,3',4'-TCB which possesses both PB- and MC-like inducing ability of liver enzymes are metabolized by one or more kinds of P450 isoform induced by PB and MC.

Animals↗

Induction of sister chromatid exchanges in cultured human lymphocytes with methylsulphonyl PCB congeners.

Methylsulphonyl polychlorinated biphenyls (MSF-PCBs) have already contaminated at relatively high concentration in the lungs and blood of Yusho patients and healthy Japanese people. Therefore, we should give due attention to their biological and toxicological effects to man. In this study, in order to evaluate S-dependent genotoxicity of five MSF-PCB congeners, namely, 3-MSF-4,5,3',4'-tetrachlorobiphenyl (TCB), 3-MSF-4,5,2',3'-TCB, 3-MSF-2,5,2',4',5'-pentachlorobiphenyl (PenCB), 4-MSF-2,5,2',3',4'-PenCB and 4-MSF-2,5,2',3',5',6'-hexachlorobiphenyl (HCB), we have examined their effects on the induction of sister chromatid exchanges (SCEs), which has been frequently used to estimate the dose of S-dependent clastogens, in cultured human lymphocytes in the absence or presence of 2,3,4,7,8-pentachlorodibenzofuran (PenCDF), 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) or 3,4,5,3',4'-pentachlorobiphenyl (Co-PenCB) and the following results were obtained. 1) 4 x 10(-5)M, 7,8-benzoflavone (ANF) and two of the five MSF-PCB congeners, namely, 3-MSF-2,5,2',4',5'- and 4-MSF-2,5,2',3',4'-PenCB at respective doses of 5.2 and 5.8 ppm, which were about 35,000 times higher than the concentrations in the lungs and adipose tissue of healthy Japanese people, significantly enhanced the frequency of SCEs. 2) In the simultaneous treatment of one of the five MSF-PCB congeners and PenCDF (3.9 ppb), TCDD (1.5 ppb) or Co-PenCB (8.8 ppb), the combination of 3-MSF-4,5,3',4'-TCB (6.8 ppb) or 4-MSF-2,5,2',3',5',6'-HCB and one of the three highly toxic chemicals significantly promoted the formation of SCEs. We have already studied whether these MSF-PCBs are non-S-dependent genotoxic compounds or not and have obtained the results that they seemed not to be or very weak ones. Therefore, based on the results of this and our former studies, the five MSF-PCB congeners examined are considered rather potent S-dependent genotoxic chemicals than non-S-dependent ones.

Cells, Cultured↗

Simultaneous detection of multiple STR loci on sex chromosomes for forensic testing of sex and identity.

The forensic usefulness of X and Y chromosomal STR loci has recently been demonstrated. One quadruplex-PCR, using 2 X- and 2 Y-STRs (STRX1/HPRTB and DYS390/ DYS393), and 2 duplex-PCRs, each using an X- and a Y-STR (ARA/DYS390 and ARA/DYS393), and detection of PCR products by using an automated DNA sequencer are reported herein. This approach allows us to determine not only the sex of the donor of a sample, but also the X- and/or Y-STR genotypes of the sample. A male biological specimen yields 4 amplified products in quadruplex-PCR and 2 amplified fragments in duplex-PCRs, whereas a female biological specimen yields only 2 amplified fragments of X-STR in quadruplex-PCR and one fragment, also of X-STR, in duplex-PCRs. Our study thus provides useful information for many activities in forensic practice, such as identity testing, paternity testing, especially of deficiency cases, compilation of population data, and sex determination of a biological sample from a single PCR.

Alleles↗

Liver and kidney function in Japanese patients with maturity-onset diabetes of the young.

OBJECTIVE: Heterozygous mutations in the transcription factors hepatocyte nuclear factor (HNF)-1 alpha, HNF-1 beta, and HNF-4 alpha are associated with maturity-onset diabetes of the young (MODY) and are believed to cause this form of diabetes by impairing pancreatic beta-cell function. The HNFs also play a central role in the tissue-specific regulation of gene expression in liver and kidney, suggesting that patients with MODY due to a mutation in HNF-1 alpha, HNF-1 beta, or HNF-4 alpha may exhibit abnormal liver or kidney function. Here, we have examined liver and kidney function in a series of Japanese patients with HNF-4 alpha/MODY1, HNF-1 alpha/MODY3, and HNF-1 beta/MODY5 diabetes. RESEARCH DESIGN AND METHODS: Clinical and biochemical data were obtained from Japanese subjects with HNF-1 alpha, HNF-1 beta, and HNF-4 alpha diabetes. The clinical data included information on BMI, age at diagnosis, current treatment, and the presence and nature of any complications. The biochemical studies examined liver and kidney function and included measures of alanine and aspartate aminotransferase, gamma-glutamyl transpeptidase, blood urea nitrogen, creatinine, uric acid, total and HDL cholesterol, triglycerides, and 17 serum proteins. RESULTS: The present age and duration of diabetes were similar in patients with HNF-1 alpha, HNF-1 beta, or HNF-4 alpha diabetes, as was the age at diagnosis of diabetes in the youngest generation. All subjects were lean. Of the subjects with HNF-1 alpha and HNF-4 alpha diabetes, 50% were treated with insulin, as were all three subjects with HNF-1 beta diabetes. Retinopathy was present in patients with each form of diabetes. None of the subjects with HNF-4 alpha diabetes had evidence of nephropathy, whereas 36% of the patients with HNF-1 alpha diabetes and 100% of those with HNF-1 beta diabetes showed diminished kidney function. The three subjects with HNF-1 beta diabetes also had abnormally high serum creatinine, uric acid, and blood urea nitrogen levels, which are consistent with impaired kidney function, and one of seven subjects with HNF-1 alpha diabetes had a mild elevation in creatinine and blood urea nitrogen levels. These values were within the normal range in the three patients with HNF-4 alpha diabetes. Although the HNFs play a role in regulating the expression of the genes for most, if not all, serum proteins, there was no decrease in the levels of any of the 17 serum proteins examined, and most were within or slightly above the normal range. Lipoprotein(a) [Lp(a)] levels were elevated in the three patients with HNF-4 alpha diabetes and in one patient with HNF-1 beta diabetes, and in a second patient with HNF-1 beta diabetes, Lp(a) was at the upper limit of normal. CONCLUSIONS: The results indicate that as in white patients, MODY resulting from mutations in the HNF-1 alpha, HNF-1 beta, and HNF-4 alpha genes in Japanese patients may be a severe disease similar to classic type 2 diabetes. In addition, they suggest that patients with HNF-1 beta diabetes may be characterized by diminished kidney function and perhaps abnormal liver function. Further studies are needed to determine whether tests of liver and kidney function will be useful in the diagnosis and subclassification of MODY.

Adolescent↗

Comparative study of MR myelography and conventional myelography in the diagnosis of lumbar spinal diseases.

The purpose of this study was to investigate the usefulness of magnetic resonance (MR) myelography by comparing it with conventional myelography. MR myelography was performed on 40 patients (24 men and 16 women; mean age, 44 years) with lumbar spinal diseases. MR myelography was imaged three-dimensionally by fast spin-echo technique with fat suppression and maximum intensity projection. We assessed both the ability to provide the images of nerve root sheaths and the similarity of the findings to conventional myelography. Satisfactory images of nerve root sheaths in the lumbar region were provided almost equally by MR and conventional myelography. The images obtained by MR myelography were similar in quality to those obtained by conventional myelography. When combined with some other conventional MR imaging techniques, this diagnostic technique of imaging lumbar spinal diseases is further enhanced.

Adolescent↗

Selectivity of electron-donor- and electron-acceptor-bonded silica packing materials for hydrophobic environmental contaminants in polar and non-polar eluents.

Electron-acceptor-bonded stationary phases, 2-(nitrophenyl)ethylsilyl (NPE) and 3-(p-nitrophenoxy)propylsilyl (NPO), and electron-donor-bonded phases, 3-(N-carbazolyl)propylsilyl (CZP), 2-(1-pyrenyl)ethylsilyl (PYE), and 5-coronenylpentylsilyl (COP), were prepared from silica particles and their selectivities were examined in both polar and non-polar solvents for specific isomers of polychlorodibenzo-p-dioxins (PCDDs), hexachloronaphthalenes (HxCNs) and planar and non-planar polychlorobiphenyl (PCB) congeners. Although no single stationary phase was able to separate all the isomer pairs that are coproduced during the synthesis of the PCDDs and HxCNs, pairs can be separated by selecting a suitable stationary phase and solvent. The separation of mixtures of PCDD isomers were found to be most successful with PYE and NPO phases, which yielded the opposite elution orders for each isomer pair that is produced as a mixture. Similar results were obtained for the HxCN isomers that were separated on PYE and CZP phases. The COP phase provided easier separation of non-ortho-substituted and mono-ortho-substituted PCBs from the other PCBs based on the planarity than PYE phase.

Chromatography, High Pressure Liquid↗

Nasopharyngeal colonization with Haemophilus influenzae type b among infants and children in Japan.

Healthy carriers of Haemophilus influenzae type b (Hib) play an important role in the spread of invasive Hib disease. The aim of the present study was to estimate Hib colonization among infants and children in Japan. Specimens from throat and nasopharyngeal cultures were obtained by thorough swabbing of both tonsils and the posterior pharynx. Specimens were inoculated on Hib antiserum agar. This was prepared with Levinthal base and Hib antiserum. Conventional methods were used concomitantly. Four of 474 infants from 1-48 months of age (0.84%) had Hib cultured from their nasopharynx. The carriage rate in 1-12 months old infants was 0.62% (2/322 cases), and that in 13-48 month old children was 1.32% (2/152 cases). Five of 167 (3.0%) 13-year-old children, and five of 154 (3.2%) 9-year-old children were asymptomatic carriers. Thirty-five of 104 household contacts of a patient with invasive Hib disease (33.6%) had Hib colonization. The carriage rate in healthy Japanese children may not be different from that in the USA prior to the availability of the conjugate Hib vaccine. The Hib carriage rate in household contacts of patients with invasive Hib disease was higher than in healthy children (P < 0.005). Our results suggest the possibility of an outbreak of invasive Hib disease in Japan.

Adolescent↗

New successful treatment with disinfectant for atopic dermatitis.

For the treatment of atopic dermatitis, a variety of therapies are used including folk medicine. At present, there is no single treatment which is effective to cure the symptoms of atopic dermatitis completely in all patients. We are drawing attention to the high isolation rate of Staphylococcus aureus when starting disinfectant treatment combined with topical steroid therapies for the purpose of killing S. aureus. As a result, we examined many patients in whom almost a complete remission was obtained even after short periods of therapy, though it had been difficult to obtain improvement by conventional treatments. In many patients, IgE values and reagin antibody titer decrease dramatically soon after starting treatment. As a disinfectant, 10% povidone-iodine solution was used. We investigated also the effect of iodine contained in the povidone-iodine solution on the thyroid gland.

Administration, Topical↗

[An autopsy case of pulmonary thromboembolism: a delayed complication of a victim by traffic accident].

A 35-year-old male had fallen down from his motor bike caused a fracture of the medial condyle of the right tibia and the acromion of the left scapula. He had received screw fixed operation for his right tibia and had been in the hospital for one month. He died suddenly in the hospital and was autopsied to find out the cause of death. On external examination, it was found that affected leg was bruised and swollened. Partially organized thrombosis in the posterior tibial vein was revealed and bilateral pulmonary arteries were packed with reddish massive thromboemboli. The causes of the posterior tibial vein thrombosis were intimal damage and decreased venous flow, attributable to bed rest for long days and external fixation of the lower extremity. We presumed that direct cause of death was a large thromboemboi which were migrated from posterior tibial vein and lodged at the bifurcation of the pulmonary artery causing obstruction of pulmonary blood flow. Finally we opened the direct cause of death was pulmonary thromboembolism due to leg injury. Clinical instability may hinder the diagnostic elevation and delay therapeutic intervention, it is the most often happened in sudden death cases within the hospital. The importance of early diagnosis and early prevention is underscoped, because the symptoms of pulmonary thromboembolism are not always produced. To prevent fatal pulmonary thromboembolism in trauma, prophylactic use of low-dose heparin therapy and/or the use of oral anticoaglant (warfarin) have been effective and recommended in USA and Western Europe. The routine use of these therapy has not yet been spread in Japan, because the incidence is still apparently lower than in USA and Western Europe as well as the risk of hemorrhage. If the victims of road traffic accident receive the prophylactic use of low-dose heparin and/or oral anticoaglant (warfarin), it might be an effective way to reduce mortality.

Accidents, Traffic↗

[The condition of PCBs and PCDFs in the blood of Yusho patients 20 years after the onset].

Blood samples of Yusho and control persons were analyzed for individual congeners of PCDDs, PCDFs, and PCBs by high resolution GC/MS. Concentrations of 2,3,4,7,8-penta-CDF, 1,2,3,4,7,8-hexa-CDF and 2,3,3',4,4',5-hexa-CB in Yusho blood were up to 56 times higher than the corresponding concentrations in the control blood. These high concentrations have persisted for 23 years after the incident. Concentrations of 3,3',4,4',5-penta-CB and 2,3',4,4',5-penta-CB in some Yusho blood were lower than the control blood. In Yusho blood, 2,3,4,7,8-penta-CDF contributed the highest toxicity (TEQ 77-248 ppt in lipid) among the congeners determined and toxic contribution of PCDFs was very large (41-77%) in the chlorinated pollutants. Thirty PCB congeners were identified in the blood of Yusho patients in 1996 by GC/MS. The average total PCB concentration in Yusho blood were 4.9 times higher than that of the controls. Characteristic PCB congeners in Yusho patients were 2,2',3,4,4',5-hexa-CB, 2,3,3',4,4',5-hexa-CB and 2,3,3',4,4',5'-hexa-CB and their concentration ratios to the controls were 8-19.

Adult↗

Direct cardiotoxic effects of cocaine and cocaethylene on isolated cardiomyocytes.

We investigated the cardiotoxic effects of cocaine and cocaethylene on the Ca2+ flux responsible for excitation-contraction coupling in isolated ventricular rat myocytes. We simultaneously measured intracellular Ca2+ transients and cell length in isolated cardiac myocytes loaded with a fluorescent Ca2+ indicator, indo-1, during electrical field stimulation at 1 Hz. The cell length was estimated by video dimension analysis. We also measured the activities of Ca2+ ATPase and Ca2+ release channels of cardiac sarcoplasmic reticulum membrane vesicles. Both cocaine and cocaethylene produced significant decreases in both peak intracellular Ca2+ and the cell-contraction rate in a dose-dependent manner. The K0.5 for the reduction of peak intracellular Ca2+ was 157.5 microM for cocaine, but 90.0 microM for cocaethylene. Both cocaethylene and cocaine inhibited neither Ca2+ ATPase nor Ca2+ release channel activity. These results demonstrate that cocaethylene has a more potent direct negative inotropic action on cardiomyocytes, without preventing Ca2+ flux through the cardiac sarcoplasmic reticulum membrane.

Animals↗

Intractable wheezing in infants with nasopharyngeal reflux.

Two infants with intractable wheezing and moist cough were referred to Chiba Municipal Kaihin Hospital. Their symptoms were persistent even after the usual treatment for respiratory disease. No definite etiological agents were detected. They usually gagged while feeding and barium swallow tests revealed nasopharyngeal reflux and cricopharyngeal incoordination. One of the patients had remarkably high titers of IgE and IgE RAST of cow's milk before she received treatment with thickened formula. She also had peripheral eosinophilia and nasal eosinophilia. These findings were thought to be caused by nasopharyngeal reflux. Four months after therapy commenced, those titers and symptoms were greatly reduced. The clinical and roentgenographic findings in these infants, and their response to therapy, strongly support a causal relationship between nasopharyngeal reflux and wheezing. Therefore, nasopharyngeal reflux should be considered when a baby has intractable wheezing, even when there is no developmental problem.

Deglutition↗