Cloning and Sequencing of an MPB70 Homologue Corresponding to MPB83 from Mycobacterium bovis BCG

MPB70 is secreted in high concentrations by Mycobacterium bovis BCG substrain Tokyo (BCG Tokyo), but little by substrains Pasteur (BCG Pasteur) and M. tuberculosis . The gene encoding a MPB70 homologue secreted by BCG Tokyo was found at the upstream region of the gene encoding MPB70, with approximately 2.3 kilobase pairs (kbp) spacing: the same gene was also found in BCG Pasteur. This gene was cloned and sequenced from BCG Tokyo. The DNA sequence which contained a 663 base pair (bp) open reading frame beginning at position 1 and ending with a TAA codon at position 661 was found. Its theoretical molecular mass was calculated to be 22.068 kDa. This gene was highly homologous to the coding region of mpb70 and the deduced amino acid sequence was very similar to MPB83 reported by Harboe et al . It was speculated that the gene the authors characterized probably corresponded to the mpb 83 gene.     

Factors useful in predicting the response to interferon therapy in chronic hepatitis C

To determine how various factors influence the response to interferon (IFN) therapy, we retrospectively studied 157 consecutive Japanese patients with chronic hepatitis C who received various treatment schedules of IFN. They were divided into two groups on the bases of outcome. One group was comprised of 65 patients who achieved a sustained normalization of serum alanine aminotransferase (ALT) levels for at least 6 months after treatment, while the other group was comprised of 84 patients with persistent elevation of serum ALT levels, despite treatment. Genotyping of hepatitis C virus (HCV) was done by polymerase chain reaction (PCR) with genotype specific primers, analysing the variations in nucleotide sequence within the NS 5 region of the HCV genome, namely genotypes PT, K1, K2a and K2b. We then used a multivariate analysis to determine the factors related to mode of treatment, patient characteristics and HCV genotype in relation to the response to IFN therapy. Of the 16 factors analysed, the HCV genotype (genotype K2a or K2b, P < 0.0008), treatment schedule (intermittent administration following a daily schedule, designated as combined schedule, P > 0.0014) and liver histology just before treatment (chronic persistent hepatitis or mild chronic aggressive hepatitis, P < 0.0324) were the most strongly correlated with a normalizing response to IFN therapy. These results suggest that not only are the IFN treatment schedule and patient profile significant, but the properties of the virus also influences the response. However, as the IFN treatment schedule is the only changeable factor, it should be designed to maximize the benefit of IFN therapy.     

Genotypic subtyping of hepatitis C virus

Four subtypes of hepatitis C virus (HCV), Pt(I), K1(II), K2a(III) and K2b(IV), have been suggested based on the nucleotide sequences of the non-structural (NS) 5 region. A fifth subtype from Japanese patients, Tr(V), which shows a less than 68% homology in nucleotide sequence when compared with other subtypes has been identified. A one-step method which enables a quick determination of subtype using polymerase chain reaction with a mixed primer set deduced from the sequence of each subtype has been developed. Using this technique, the subtypes of 418 out of 478 Japanese patients (87.4%) were determined. The incidence of each subtype in Japan was as follows: K1(II), 307 (73.4%); K2a(III), 74 (17.7%); K2b(IV), 28 (6.7%); and Tr(V), 3 (0.7%). This one-step subtyping technique should be useful for studying the epidemiology or biology of the HCV.     

Prevalence of gastroesophageal reflux disease and gastroesophageal reflux disease symptoms in Japan

Background and Aim:  Epidemiological studies have shown that 10–48% of people in developed countries have gastroesophageal reflux disease (GERD) symptoms such as heartburn and acid regurgitation. The present study aimed to examine the prevalence of GERD symptoms and GERD in Japanese subjects.
Methods:  A cross-sectional study of Japanese subjects who visited a clinic for a routine health check up was carried out. Subjects were asked to fill out a self-report questionnaire. GERD was defined as the presence of heartburn and/or acid regurgitation at least twice per week.
Results:  Of the 6035 eligible subjects, 2662 (44.1%) reported having had heartburn and/or acid regurgitation during the past year: 124 (2.1%) daily, 275 (4.6%) twice per week, 773 (12.8%) twice per month and 1490 (24.7%) less than twice per month. Three hundred and ninety-nine (6.6%) subjects were diagnosed as having GERD and there was no relationship between the prevalence of GERD and either sex or age. The prevalence of bothersome GERD symptoms was significantly higher in subjects with GERD than in those without GERD.
Conclusion:  Approximately 6.6% of Japanese have GERD and most persons with GERD described heartburn or acid regurgitation as bothersome.     

Alpha-blocker test: Alternative to pressure–flow study of bladder outlet obstruction and detrusor contractility in patients without an enlarged prostate

OBJECTIVES: We investigated whether the cause of urinary disturbance in men with a prostate volume < or =20 mL can be determined by analyzing the efficacy of alpha1-adrenoceptor antagonist (alpha-blocker) treatment. METHODS: Thirty-five men who were >50 years of age, with an International Prostate Symptom Score (IPSS) > or =8 points, a quality of life (QOL) index > or =2 points and a prostate volume 20 mL served as controls. The alpha1-adrenoceptor antagonist tamsulosin was administered at a dose of 0.2 mg/day for 4 weeks. Results for the IPSS, QOL index, free flowmetry and pressure-flow studies were obtained before and after tamsulosin administration. RESULTS: In both groups, tamsulosin improved the IPSS and QOL index and the bladder outlet obstruction index (BOOI) was lowered without reducing the bladder contractility index (BCI). No parameter showed a significant difference in treatment efficacy between the two groups. In the non-enlarged prostate group, both the pretreatment BOOI and BCI correlated with the efficacy of treatment in improving maximum flow rate (Qmax). In the enlarged prostate group, BOOI and BCI did not correlate with Qmax. When Qmax was improved by > or =3.5 mL/s, the positive predictive value for both pretreatment BOOI >40 and BCI >100 was 100% in the non-enlarged prostate group. CONCLUSIONS: The alpha-blocker test is one method to assess the presence of bladder outlet obstruction and the state of detrusor contractility in men without an enlarged prostate.     

Gas chromatographic-mass spectrometric metabolic profiling of patients with fatal infantile mitochondrial myopathy with de Toni-Fanconi-Debré syndrome

The metabolic profiles of three patients with fatal infantile mitochondrial myopathy with de Toni-Fanconi-Debré syndrome were studied by simultaneous analysis, after urease treatment of urinary organic acids, carbohydrates, polyols and amino acids using gas chromatography/mass spectrometry (GC/MS). All three patients persistently showed lactic aciduria, phosphaturia, glucosuria and generalized amino aciduria. This abnormal urinary metabolic profile was observed before the onset of any clinical symptoms, indicating that chemical diagnosis may be done presymptomatically. In one patient, the concentration of lactate increased in parallel with the severity of the clinical condition, whereas the urinary levels of 3-hydroxybutyrate, amino acids and glucose fluctuated and showed only a general tendency to increase with the clinical course. The above results suggest that simultaneous GC/MS analyses, without fractionation, of urinary metabolites facilitate not only the early chemical diagnosis either before or after the first onset, but also follow-up studies, providing an important index for the evaluation of the severity and clinical course in patients with this disorder.     

C5b-8 Step Lysis of Swine Endothelial Cells by Human Complement and Functional Feature of Transfected CD59

The authors established several swine endothelial cell (SEC) lines expressing human CD59 by transfection of cDNA, and assessed the function of the transfectant molecules in comparison with those of membrane cofactor protein (MCP) and decay-accelerating factor (DAF) in an in vitro hyperacute rejection model of swine to human discordant xenograft. At the usual expression rate, DAF and MCP protected SEC from human complement mediated cell lysis, but CD59 did not block human complement attack on SEC. However, CD59 protects SEC from cell lysis when sufficiently expressed as in human umbilical vein (HUVEC). The authors examined why CD59 needed so many molecules to protect human complement-mediated SEC lysis and found that SEC underwent lysis by human C5b-8. The degree of C5b-8 step lysis of SEC was approximately 70% of the total activity (C5b-9). Additionally, CD59 protected human complement activation less efficiently at the C5b-8 step than at the C9-step. Therefore, to overcome human complement mediated SEC lysis, C8 activity must be inhibited by dense expression of CD59.     

Newly Developed Miniature Ultrasonic Probe,Basic and Clinical Study

The authors evaluated a new device-the miniature ultrasonic probe (3.4mm in the outer diameter, radial scan type, 7.5MHz, Olympus XMP-02) from both its basic and clinical aspects using fresh resected gastric specimens with water immersion The ultrasonic resolution of this new probe was demonstrated to be best when the probe-object distance was 1.0–1.5cm and the angle of inclination was0–20*. Practical use of the probe with a smaller diameter is limited in the imaging range of horizontal directions; only 0.7cm right and left from the center receiving ultrasonic beams at a right angle. Although the ultrasonic resolution of this new device is inferior to that of conventional ultrasonography instruments, it can be applied to stenosed lesions and possibly to the bile duct or pancreatic duct regions in the future. This device, with further improvement, is expected to become much more useful for clinical application.     

Topical Application of an Anti-tumor Agent Through a PTBD Catheter for Management of an Incurable Malignant Biliary Obstruction —a PTBD Option—

Abstract: The treatment of a 69-year-old woman with obstructive jaundice due to the direct invasion of metustutic lymph nodes by gastric cancer at the porta hepatis is reported. In this case topical infusion chemotherapy via a percutaneous biliary drainage (PTBD) catheter with Cis-Diammine Dichloro-Platinum (CDDP) plus Adriamycin (ADM) was found to be an effective treatment to achieve internal biliary drainage with the aid of a percutuneous transhepatic cholangioscopy. This result, in our opinion, suggests that the topical use of anti-tumor agents via a PTBD catheter might be considered an option for the treatment of patients with incurable malignant biliary obstruction.