Questionnaire of cryopreserved allograft valves in Kyushu and Yamaguchi area

Japanese Society of Cryopreserved Thoracic Tissue Implantation was recently set up. Cryopreserved allograft valves are about to pervade in Japan. To clarify the interest, demands and supply of cryopreserved allograft valve in the area of Kyushu and Yamaguchi, we performed questionnaire investigation regarding this issue. Collection rate of this questionnaire was 87.5% (35/40 hospitals). Ninety-seven percent of the hospitals answered that they were interested in cryopreserved allograft. Ninety-four percent of the hospitals answered that they did not ethically hesitate to use cryopreserved allograft. Ninety-one percent of the hospitals hoped to perform cardiac surgery using allograft aortic valve if allograft is available. With respect to securing donors of allograft, two-third of the hospitals did not decide whether they could be involved in obtaining donors of allograft. As the number of the cadaver kidney donors is about 20 per year in the area of Kyushu and Yamaguchi, shortage of donors of allograft valve is anticipated.     

Lysozyme requires fluctuation of the active site for the manifestation of activity

Mutations around His15 which lie far away from the active site,stimulated glycol chitin activity of lysozyme at physiologicaltemperature. Del-Argl4Hisl5 lysozyme, a mutant lysozyme whoseArgl4 and Hisl5 were deleted together, and has the highest activityamong these mutant lysozymes, had a similar binding abilityto a trimer of N-acetyl-glucosamine, a substrate analogue, relativeto native lysozyme. This suggests that the increased activitywas due to an increased k_cat in the catalysis reaction. TheH-D exchange rate of the N-1 proton in the Trp63 which is locatedin the active site cleft, was enhanced in the Del-Argl4Hisl5lysozyme, while 2-D proton NMR analysis revealed no conformationalchange around Trp63. We conclude that some sort of fluctuationat the active site might be required for the manifestation ofactivity. This theory is supported by the finding that the Del-Argl4Hisl5lysozyme showed a shift in temperature dependency of activityto lower temperatures compared with that of native lysozyme.     

An improved method for preparing lysozyme with chemically 13C-enriched methionine residues using 2-aminothiophenol as a reagent of thiolysis

Jones et al. have reported that the epsilon-carbons of methionine residues in myoglobin can be enriched with stable isotope (13C) in two steps, i.e., methylation of methionine residues with 13CH3I in the protein and thiolysis using dithiothreitol [Jones, W.C., Rothgeb, T.M., and Gurd, F.R.N. (1976) J. Biol. Chem. 251,7452-7460]. Using their method, we failed to prepare active lysozyme in which the epsilon-carbons of methionine residues are enriched with 13C, because many side reactions took place under the thiolysis condition (pH 10.5, 37 degrees C). When we employed 2-aminothiophenol as a reagent for thiolysis, the reduction proceeded under a weakly acidic condition to afford fully active lysozyme, in which the epsilon-carbons of two methionine residues were enriched with 13C, in a 30% yield. Analysis of the 13C-edited NOESY spectra of 13C-enriched methionine lysozyme in the absence and presence of a substrate analogue indicated the occurrence of conformational change around Met 105 in lysozyme.     

Feasibility of adjuvant chemotherapy for breast cancer patients

The prevalence and natural history of severe proteinuria in mild to moderate hypertension are not completely defined. We screened 1635 men with a history of hypertension and randomized 1292 with untreated diastolic blood pressure (DBP) 95-109 mmHg to single-drug treatment with either hydrochlorothiazide, atenolol, captopril, clonidine, diltiazem-SR, prazosin, or placebo in a double-blind prospective trial. Twenty-seven of 1635 patients (1.7%) satisfying clinical criteria for primary hypertension were found to have developed proteinuria > 1000 mg/24 hours and were removed from the study. Follow-up data were obtained on 19 of these 27 patients. One patient was found to have focal segmental sclerosis and progressed to end-stage renal disease. Three other patients developed severe (serum creatinine > 3.5 mg/dl) chronic renal failure (one with diabetic nephropathy), one progressed from serum creatinine 1.4 to 2.2 mg/dl, but 14 of the 19 remained with stable serum creatinine < 2.0 mg/dl on follow-up for 6-9 years. Data were available for 1076 of 1155 (93%) treated study patients at end titration, 522/600 (87%) at one year and 322/444 (73%) at two years. There were significant associations for proteinuria with obesity and higher systolic blood pressure. There was a trend toward significant difference in mean 24-hour protein excretion rates at baseline between black (127 mg) and white (139 mg) patients (p = 0.07). There were no statistically significant changes in urinary protein excretion/24 hours between or within the different treatment groups (including placebo). Eighteen patients were removed from the study during the active treatment phase for proteinuria > 1000 mg/24 hours: hydrochlorothiazide 4, placebo 3, diltiazem 3, prazosin 3, atenolol 2, clonidine 2, and captopril 1. We conclude: (1) the prevalence of severe (> 1 g/24 hours) proteinuria in the hypertensive population is significant but does not necessarily imply a poor prognosis; (2) mean 24-hour urinary protein excretion rates did not vary in response to the different classes of antihypertensive drugs; and (3) there was no drug-specific increase in proteinuria detected in this study.     

Assessment and treatment of liver disease in Japanese haemophilia patients

OBJECTIVES: To examine the efficacy of various parameters obtained by transrectal ultrasonography (TRUS) as predictors of the onset of acute urinary retention in patients with benign prostatic hyperplasia (BPH). METHODS: From April 1993 to March 1997, 331 men aged 51 to 84 years with symptoms of BPH were enrolled in this study. Among them, 64 patients presented to our clinic because of acute urinary retention. TRUS was used to calculate the transition zone (TZ) volume, the transition zone index (TZ index = TZ volume/total prostate volume), the total prostate volume, and presumed circle area ratio (PCAR). To compare the usefulness of the various indices, the area under the receiver-operator characteristic (ROC) curve was calculated for each index. RESULTS: There were significant differences in the American Urological Association (AUA) symptom score, total prostate volume, TZ volume, TZ index, and PCAR between patients with and without acute urinary retention, but no significant differences in age and quality of life score. In patients with acute urinary retention, the area under the ROC curve was 0.924 for the TZ index, 0.834 for the TZ volume, 0.753 for the PCAR, 0.684 for the total prostate volume, and 0.628 for the AUA symptom score. CONCLUSIONS: The TZ index is an accurate predictor of acute urinary retention in patients with BPH and may be a useful method for deciding between surgical intervention and antiandrogen treatment.     

Analysis of the stabilization of hen lysozyme by helix macrodipole and charged side chain interaction

In the N-terminal region of the alpha-helix of the c-type lysozymes, two Asx residues exist at the 18th and 27th positions. Hen lysozyme has Asp18/Asn27 (18D/27N), and we prepared three mutant lysozymes, Asn18/Asn27 (18N/27N), Asn18/Asp27 (18N/27D), and Asp18/Asp27 (18D/27D). The stability of the wild-type (18D/27N) lysozyme supported the existence of a hydrogen bond between the side chain of Asp18 and the amide group at the N1 position in the alpha-helix, while the stability of the 18N/27D lysozyme supported the presence of the capping box between the Ser24 (N-cap) and Asp27 residues. Although electrostatic repulsion was observed between Asp18 and Asp27 residues in 18D/27D lysozyme, the dissociation of each residue contributed to stabilizing the B-helix in 18D/27D lysozyme through hydrogen bonding and charge-helix macrodipole interaction. This is the first evidence that two neighboring negative charges at the N-terminus of the helix both increased the stability of the protein.     

High pressure NMR study of a small protein, gurmarin

The incremental elastic modulus of the common carotid and radial arteries is increased in patients with end-stage renal disease (ESRD), independently of blood pressure, wall stress, and the presence of atherosclerotic alterations. Whether biochemical factors may be involved in the arterial changes and related to renal dysfunction remain largely ignored. To assess this question, we measured aortic (carotid-femoral), upper-limb (carotid-radial), and lower-limb (femoral-tibial) pulse wave velocity (PWV) in 74 ESRD patients undergoing hemodialysis in comparison with 57 control subjects similar in age, sex ratio, and mean blood pressure. We evaluated arterial blood pressure by sphygmomanometry, aortic calcifications and cardiac mass by echography, and routine biochemical parameters, total plasma homocysteine, and plasma endothelin levels by standard techniques. In the population of patients with ESRD, on the basis of multiple stepwise regression analysis, aortic PWV was positively and independently correlated with systolic blood pressure (P<.0001), age (P<.0001), prevalence of aortic calcification (P=.0004), and the prevalence of diabetes mellitus (P=.0043). Upper-limb PWV was influenced exclusively by mean blood pressure (P<.0001). Lower-limb PWV was positively and independently correlated with plasma total homocysteine (P=.0004) and plasma endothelin (P=.0187) only. At any vascular site, PWV was not independently correlated with tobacco consumption; plasma levels of cholesterol, triglyceride, fibrinogen, or hemoglobin; body mass index; or the presence of bilateral nephrectomy. Finally, plasma homocysteine was independently correlated with cardiac mass (P=.0022). This study provides evidence that in ESRD patients, the stiffness of the arterial wall and cardiac mass are strongly influenced by biochemical factors related to the kidney alterations and are independent of age and blood pressure level. Increased plasma endothelin and homocysteine may be specifically involved in the vascular damage of lower limbs.     

Properties of polarization noise caused by external disturbances insingle-mode fiber transmission systems

The properties of polarization noise caused by external disturbances in intensity modulation/direct detection transmission systems that have polarization-dependent optical devices are analyzed. It is experimentally shown that the noise consists of additive noise and multiplicative noise. The multiplicative noise directly disturbs signals, even if the external disturbance is of a low frequency. The signal to polarization noise ratio easily reaches the ratio at which flickers obstruct the picture in an analog TV transmission system. The polarization noise caused by external disturbances is also discussed. It is shown that disturbances to the fiber can make the maximum noise n ² times greater than the noise caused by a single disturbance, where n is the number of disturbances