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Biomedical subjects

A Kuroda

Publications and source records attributed to A Kuroda.

At least 109 records · Page 6Linked to original sources

Molecular cloning and sequencing of a major Bacillus subtilis autolysin gene.

A major Bacillus subtilis 168S autolysin (N-acetylmuramoyl-L-alanine amidase [EC 3.5.1.28]) was purified and then cleaved with cyanogen bromide. The N-terminal amino acid sequence of one of the resultant peptides was determined in order to make synthetic oligonucleotides. A 2.5-kb EcoRI fragment was cloned into Escherichia coli JM109 and detected by colony hybridization by using the oligonucleotides as probes. Sequencing of the insert showed the presence of an open reading frame (designated cwlB), starting at a UUG codon, which encodes a polypeptide of 496 amino acids with a molecular mass of 52,623 Da. CWLB had a presumed signal peptide which is processed after Ala at position 24. Insertional inactivation of the cwlB gene of the B. subtilis chromosome led to an approximately 90% decrease in the total cell wall hydrolytic activity of stationary-phase cells and extraordinary resistance to cell lysis, even after 6 days of incubation at 37 degrees C. No apparent changes in cell morphology, motility, competence, sporulation, or germination were observed.

Amino Acid Sequence↗

[A case of malignant pheochromocytoma of the urinary bladder].

A 31-year-old man was admitted to our hospital because of miction pain and gross hematuria in August, 1988. Cystoscopic examination revealed a solid mass in the bladder. The histological diagnosis was pheochromocytoma of urinary bladder by biopsy. He had a history of syncope on urination several times. Abnormal elevation of serum noradrenaline was observed in hormonal studies and abnormal accumulation was seen in the bladder by 131I-MIBG scintigraphy. Total cystectomy, lymphadenectomy and urinary diversion were performed in November, 1988. Pathological diagnosis was pheochromocytoma of the urinary bladder with metastasis to both iliac lymph nodes.

3-Iodobenzylguanidine↗

Massive pancreatic pleural effusion: pathogenesis of pancreatic duct disruption.

Massive pancreatic pleural effusion is believed to be caused by pancreatic duct disruption. To elucidate the cause of the duct disruption, pancreas specimens resected from three cases of massive pancreatic pleural effusion were investigated histopathologically. There was no evidence of either chronic or acute pancreatitis except focal fibrosis along a single dilated duct in the resected pancreas. From our study, the pathogenesis of pancreatic duct disruption is suggested as follows: (a) alcohol ingestion can induce focal acute inflammation on a single branch of the duct system and elicit protein plug formation; and (b) focal stenotic change occurs on this branch at a point near the main pancreatic duct, and transient obstruction can occur by means of the protein plugs. The "upstream" extent of this branch will then dilate until it ruptures. However, on the frequency of this atypical pancreatitis, we have no clues to mention.

Acute Disease↗

Cloning, sequencing and genetic mapping of a Bacillus subtilis cell wall hydrolase gene.

We have cloned DNA fragments from Bacillus subtilis 168S into Escherichia coli, which produced a lytic zone on an agar medium containing B. subtilis cell wall. Sequencing of the fragments showed the presence of an open reading frame (ORF) which encodes a polypeptide of 272 amino acids with a molecular mass of 29919 Da. The deduced amino acid sequence showed considerable homology with that of the cell wall hydrolase gene of Bacillus sp. (Potvin, C., Leclerc, D., Tremblay, G., Asselin, A. & Bellemare, G. (1988). Molecular and General Genetics 214, 241-248). Accordingly, the gene was designated cwlA, for cell wall lysis. The N-terminal amino acid sequence of cwlA gene product prepared from a E. coli clone was AIKVVKNLVSKSKYGLKCPN, which is consistent with that of the deduced sequence starting from Ala at second position from the initiation codon of the cwlA gene. A presumed sigma A promoter and a rho-independent terminator were found upstream and downstream of the ORF, respectively. A chloramphenicol-resistance determinant integrated into the ORF was mapped by PBS1 transduction, which indicated the gene sequence dnaE-aroD-cwlA.

Amino Acid Sequence↗

Nucleotide sequences of the Bacillus subtilis flaD locus and a B. licheniformis homologue affecting the autolysin level and flagellation.

A DNA fragment containing the flaD locus of Bacillus subtilis, which had been cloned into plasmid pAC3, was subcloned into an M13 phage and sequenced. The sequence contained five open reading frames (ORFs), of which ORF2 was the flaD gene. Unexpectedly, the sequence of the flaD locus was identical to that of sin [sporulation inhibition gene; Gaur, N. K., Dubnau, E. & Smith, I. (1986). Journal of Bacteriology 168, 860-869]. A B. licheniformis homologue (flaL) of the B. subtilis flaD locus was cloned into pUC19 and identified by colony hybridization. The B. licheniformis DNA was subcloned and sequenced. Two ORFs (ORF1, or L-ORF1; and ORF2, or flaL) were detected, encoding 58 and 111 amino acid residues, respectively. These are almost identical in length to ORF1 (D-ORF1; 57 amino acids) and flaD (111 amino acids) on the fragment of B. subtilis DNA. The overall interspecies differences between the nucleotide sequences of D-ORF1 and L-ORF1, and those of flaD and flaL, were 42% and 11%, respectively, and the differences in the predicted amino acid sequences were 50% and 7%, respectively. The regions 3' of the ORFs (flaL and flaD) in both species resemble rho-independent terminators of transcription. The characteristics of the amino acid sequences are also discussed.

Amino Acid Sequence↗

Histochemical studies on the conduction system of diabetic rat hearts.

We studied the metabolism of the conduction system and the working myocardium in diabetic rat hearts by enzyme histochemistry. The experiment was performed three weeks following the administration of streptozotocin (65 mg/kg) to male Wistar rats. The hearts were quickly excised and tissue was frozen immediately by immersion in isopentane at -30 degrees C and cut into 16 microns thick sections in a cryostat. The PAS positive reaction was increased in the conduction system compared to the working myocardium in control rats. In diabetic rat hearts, these reactions in the working myocardium and the conduction system were strongly increased compared to control hearts. Several enzyme activities, such as phosphofructokinase, pyruvate dehydrogenase, glucose-6-phosphate dehydrogenase, Na-K ATPase, were reduced in both the working myocardium and conduction system of diabetic rat hearts. These results suggest that the changes in metabolic condition also exist in the conduction system of the diabetic rat hearts as well as the working myocardium.

Animals↗

[Early changes in serum alpha-fetoprotein after chemotherapy in advanced testicular tumors].

The serum levels of alpha-fetoprotein (AFP) are known to undergo acute changes (paradoxical elevation) after effective chemotherapies for testicular germ cell tumors. In order to understand the clinical significance, the changes of the serum AFP levels after PVB therapy for testicular germ cell tumors were analyzed, and their relationship to the treatment effects was investigated in this retrospective study. Thirty-one patients with testicular germ cell tumors (11 with stage I, 7 with stage II, and 13 with stage III) which had been treated by PVB therapy were evaluated. The measurement of the serum AFP levels was performed by radioimmunoassay on Day 1 (the initiation day of the PVB therapy), thereafter on Day 8 and Day 22. The half-life (t1/2) or doubling time (t2) of the serum AFP were calculated. In patients with stage I disease, the serum AFP levels declined nearly according to its biological half-life between Day 1 and Day 22 after termination of chemotherapy (t1/2 = 6.2 days). In patients with stage II or III disease, the changes of the serum AFP levels were not consistent with the treatment results between Day 1 and Day 8. Namely, complete responders revealed a prolonged decline of the serum AFP levels (t1/2 = 63.0 days), partial responders a transient rise of the serum AFP levels (t2 = 49.5 days) and non-responders a sudden rise of the serum AFP levels (t2 = 9.9 days). On the contrary, the serum AFP levels changed in accordance with the treatment effects between Day 8 and Day 22. Early changes of the serum AFP levels were seen in patients with stage II or III disease between Day 1 and Day 8 after chemotherapy. It was indicated that the measurements of the serum AFP levels during this period were not useful for the evaluation of the treatment effects in these patients. Furthermore, it was suggested that the early changes of the serum AFP levels possibly correlate with the poor response. The mechanism of the early changes of the serum AFP was also discussed.

Adolescent↗

[Clinicopathologic study on minute endocrine tumors of the pancreas--analysis of autopsy cases].

Using 800 autopsy cases (422 men, 378 women, mean age 78.7) mainly consisting of the aged, we made a clinicopathological analysis of endocrine tumors of the pancreas. The results were: 1) Endocrine tumors and similar lesions were found in 24 cases (25 lesions). 20 lesions (20 cases) were determined as tumors and five lesions (five cases) as hyperplasia of Langerhans islet. 2) Prevalence of tumor was 10% (6/60) in the all-section group and 1.6% (12/738) in the three-section group. Grossly observed tumor was in a single case. 3) As for the size of tumors, 35% of them were 1,000 microns or smaller. All of the five cases of hyperplastic lesions were 1,000 microns, or smaller in size. 4) None of the cases with tumors and hyperplastic lesions showed symptoms of excessive hormone. 5) In immunohistochemical analysis, hormone production was observed in all of the 20 cases of tumor and multiple hormone production was found in 14 of them (70%). 5) Ductular or tubular structures were found in or adjacent to tumor in 12 cases (60%) and hyperplasia in one case (20%). 6) One lesion each of tumor and hyperplasia were found in a single case. Langerhans islets, 500 microns or larger in size, were found in three lesions of tumor (15%). Furthermore, langerhans islets with the diameter of mean + 2 SD or larger were found around five lesions of tumor (25%) and three lesions of hyperplasia (60%). Above findings suggest that endocrine tumors of the pancreas is highly prevalent and most of them do not go beyond asymptomatic tumors (non-functional in the broad sense of meaning) with no symptoms of excessive hormone though they produce some hormones. And it is indicated that some of the endocrine tumors or hyperplasias develop from totipotent stem cells of duct epithelia, and factors that promote the growth of Langerhans islets might exist.

Adenoma, Islet Cell↗

Arrhythmogenic right ventricular dysplasia with left ventricular involvement: report of a case.

A case of arrhythmogenic right ventricular dysplasia with histological abnormality of the left ventricle is reported. Right ventriculography showed severe dilatation and hypokinesis, and left ventriculography also showed slightly diffuse hypokinesis. Histological findings of both ventricles revealed myocytolysis, severe fibrosis, and marked fatty infiltrations. It is therefore important to examine histologically both the left and right ventricles for such lesions.

Adult↗

Clinicopathologic study of asymptomatic gallbladder carcinoma found at autopsy.

Of 3000 autopsy cases, consisting primarily of older patients, asymptomatic gallbladder carcinomas were found in 15 (0.5%); the incidence among all gallbladder carcinomas (80 cases) was 18.8%. In contrast to symptomatic gallbladder carcinomas, all of which were classified as Stage III (9.3%) or Stage IV (90.7%) disease, 80.0% of asymptomatic cases were classified as Stage 0 (13.3%), I (20.0%), or II (46.7%). Histologically, all the symptomatic cases of gallbladder carcinoma had invasion to the serosal or subserosal layer of the gallbladder as well as to the hepatoduodenal ligament. Lymphatic and hematogenous metastases of the symptomatic carcinomas were found in 94.4% and 64.8%, respectively. On the other hand, the carcinoma in the asymptomatic group was retained in the mucosal layer (M) in four cases, in the muscular layer (MUS) in one case, in the subserosal layer (SS) in nine cases (INF beta in seven cases and INF r in two in the mode of infiltration), and in the serosa (SE) in one case. However, cancerous invasion to the hepatoduodenal ligament or lymphatic metastasis was found in only two cases, whereas hematogenous metastasis was not found in any of them. Cancerous extension beyond the gallbladder wall was apparently rare in the cases of SS invasion, as well as in those of M or MUS invasion. It was suggested that, for radical resection of gallbladder carcinoma, it is very important to find it in an asymptomatic stage.

Adenocarcinoma↗

Characterization of human pancreatic kallikrein.

Human pancreatic kallikrein (H.Panc.K.) was purified from human pancreas by ion exchange chromatography on DEAE-cellulose, affinity chromatographies on p-aminobenzamidine Sepharose 6B and aprotinin aminocellulofine, followed by gel filtration on Sephacryl S-200. The final preparation had a specific activity of 9.2 AU/A280 (AU; amidase unit for H-Pro-Phe-Arg-MCA) and its N-terminal sequence coincided with the reported sequence for H.Panc.K.. In HPLC (gel filtration), one symmetrical peak corresponding to a molecular weight of 48,000 was obtained. In SDS-PAGE without 2-mercaptoethanol (2-ME), one band corresponding to a molecular weight of 52,000 was obtained, but with 2-ME, 2 bands, 52,000 and 30,000, were obtained. Km value for MCA was 4.9 x 10(-2) mM. Proteinase inhibitor specificities of H.Panc.K. were the same as those of human urinary kallikrein (HUK) and hog pancreatic kallikrein (hog Panc.K.), while anti-HUK antibody inhibited the activities of H.Panc.K. and HUK, but not that of hog Panc.K.. From the analysis of affinity for concanavalin A (Con A) and erythroagglutinating phytohemagglutinin (E-PHA), the carbohydrate parts of H.Panc.K. are relatively rich in biantennary complex type sugar chains with bisecting GlcNAc compared with those of human salivary kallikrein (H.Saliv.K.) and HUK.

Amino Acid Sequence↗

Biliary malignancies occurring in choledochal cysts.

Eight cases of choledochal cyst associated with biliary malignancy (gallbladder carcinoma in three and bile duct carcinoma in five) were reviewed to evaluate the roles and limitations of computed tomography (CT) (n = 8), ultrasound (US) (n = 6), cholangiography (n = 8), and angiography (n = 6). In cases of gallbladder carcinoma, both CT and US revealed mass lesions in the gallbladder consistent with cancer. CT also depicted either a mass lesion or an irregular thickened wall in all cases of bile duct carcinoma; however, US failed to demonstrate a thickened wall in one of three cases in which CT findings were positive. Cholangiography universally revealed malunion of the pancreatico-biliary duct, and the findings were suggestive of malignancy in seven cases in which CT depicted abnormalities. Angiography showed tumor stain in three of six cases. Lymph node metastases were present in four cases but were detected preoperatively in only one. One case showing a thickened bile duct wall was erroneously diagnosed as malignancy among 27 cases of choledochal cyst unassociated with biliary malignancy.

Adult↗

[Purification of human pancreatic kallikrein and organ-specificities of human glandular kallikreins].

Human pancreatic kallikrein (H. Panc. K.) was purified from human pancreas by serial liquid chromatographies. The final preparation had a specific activity of 9.2 AU/A280 (AU: amidase unit for H-Pro-Phe-Arg-MCA) and its N-terminal sequence coincided with the reported sequence determined from cloned cDNA analysis. In HPLC (gel filtration), one symmetrical peak corresponding to a molecular weight of 48,000 was obtained. In SDS-PAGE without 2-mercaptoethanol, one band corresponding to a molecular weight of 52,000 was obtained. Protease inhibitor specificities of H. Panc. K. were the same as those of human urinary kallikrein (HUK) and hog pancreatic kallikrein (hog Panc. K.), while anti-HUK rabbit antibody inhibited the activities of H. Panc. K. and HUK, but not that of hog Panc. K. From the analysis of affinity for concanavalin A and erythroagglutinating phytohemagglutinin, the carbohydrate parts of H. Panc. K. are relatively rich in bi-(or multi-) antennary complex type sugar chains with bisecting GlcNAc compared with those of human salivary kallikrein and HUK. These findings will be a help to clarify the physiological and pathophysiological roles of H. Panc. K. in the pancreas and pancreatic diseases, especially in acute pancreatitis.

Carbohydrate Conformation↗

Carcinoma of the gallbladder and extrahepatic bile duct in autopsy cases of the aged, with special reference to its relationship to gallstones.

To demonstrate correlation between occurrence of carcinoma and that of gallstone of the gallbladder and biliary tract, we reviewed the protocols of gallbladder and extrahepatic biliary duct carcinoma and cholelithiasis of 4,482 cases (male 2,237, female 2,245, mean age 77.7 yr) autopsied at the Department of Pathology, Tokyo Metropolitan Geriatric Hospital, during the 27 yr from 1960 to 1986. Gallbladder carcinoma was found in 94 cases, or 2.1%. The incidence was higher in the female than in the male (male 24, female 70, p less than 0.01). Gallstone of the gallbladder was found in 957 cases or 21.4%. In the male, incidences of gallbladder carcinoma and stone increased with age until the lower half of the ninety, whereas in the female, no such tendency was found after the sixties. Incidence of gallbladder carcinoma was significantly higher in the cases with cholecystolithiasis than in those without stone (p less than 0.01). Furthermore, the incidence of gallbladder stones in the cases with relatively early carcinoma was significantly higher than that of those without carcinoma (p less than 0.01). Cholesterol stones were more common than bilirubinate in the carcinoma patients. These results suggest the importance of cholecystolithiasis, especially that of the cholesterol stones, as a background factor of gallbladder carcinoma. Extrahepatic bile duct carcinomas were present in 33 cases or 0.7% (male 19, female 14, no sex preference). The incidence was significantly higher in the cases with stones than in those without stones of the extrahepatic bile ducts (p less than 0.01). However, the fact that small stones were found in the upstream portions of obstruction of biliary tracts and no stone was found at operations in the 11 operated cases suggested that stones may be secondary to the cancerous growth.

Adolescent↗

FR-900520 and FR-900523, novel immunosuppressants isolated from a Streptomyces. II. Fermentation, isolation and physico-chemical and biological characteristics.

FR-900520 and FR-900523, novel neutral macrolide immunosuppressants, were isolated from the cultured broth of Streptomyces hygroscopicus subsp. yakushimaensis No. 7238. Their molecular formulae were determined as C43H69NO12 and C42H67NO12, respectively. The compounds suppressed immune response in vitro. IC50 values of FR-900520 and FR-900523 for mouse mixed lymphocyte reaction were 0.55 nM and 1.6 nM, respectively. FR-900520, the major component, clearly prolonged skin allograft survival in rats.

Animals↗

FR65814, a novel immunosuppressant isolated from a Penicillium strain. Taxonomy, fermentation, isolation, physico-chemical and biological characteristics and structure assignment.

FR65814, a novel immunosuppressant, was isolated from the cultured broth of Penicillium jensenii F-2883. The structure was assigned to be 5,6-dihydroxy-4-(1,2-epoxy-1,5-dimethyl-4-hexenyl)-1-oxaspiro++ +[2,5]octane by spectroscopic analyses. The compound suppressed the immune response at low concentration. In addition, a structually related component fumagillol, a known carcinolytic agent, was also isolated and found to show immunosuppressive activity.

Animals↗