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Biomedical subjects

N Harada

Publications and source records attributed to N Harada.

At least 595 records · Page 33Linked to original sources
PubMed1985

Surgical treatment for carcinoma of the extrahepatic bile duct.

Eighty-six patients with primary extrahepatic bile duct carcinoma operated on in the Second Department of Surgery at Nagasaki University Hospital during a recent 13.5-year period were reviewed. The patients were divided into five groups depending upon the site of the tumor. The operative mortality, resectability, postoperative survival period, and five-year survival rate in each group were studied. The lower third group had the highest resectability, lowest operative mortality and longest post operative survival period. The hepatic duct, the upper third and the extended groups, however, showed extremely poor results. The pathological features of these three groups are discussed here and an operative procedure for resection of the tumor is proposed. We emphasize that the development of methods of early diagnosis is necessary, and recommend aggressive surgical treatment for tumors of the hepatic duct, upper third, and extended groups.

Adolescent↗
PubMed1985

Long-term results of surgical treatment for intrahepatic stones.

One hundred and nineteen patients with intrahepatic stones treated surgically in Nagasaki University Hospital from 1969 to 1984 were reviewed. The patients were divided into four types according to location of the stones and the presence or absence of stenotic lesions and/or localized dilatation of the intrahepatic bile ducts. Types I and II patients were treated with choledocholithotomy or choledochojejunostomy, while type III patients underwent hepatic resection and type IV patients were treated by partial hepatic resection with bilioenteric anastomosis, including extended hepatico-choledochojejunostomy. The majority of operative or early deaths belonged to type IV and residual stones were present in almost all patients. The long-term results for the 88 patients revealed that the rate of improvement was 100 per cent for type I, 87 per cent for type II, 83 per cent for type III and 84 per cent for type IV. In type IV, the most excellent results (92 per cent) were obtained by extended hepaticocholedochojejunostomy, especially with hepatectomy. It is suggested that extended hepaticocholedochojejunostomy with partial hepatic resection is a reasonable procedure for treating patients with type IV intrahepatic stones.

Adolescent↗
PubMed1985

Sex-dependent expression of mouse testosterone 16 alpha-hydroxylase (cytochrome P-450(16) alpha): cDNA cloning and pretranslational regulation.

By using both double-colony hybridization and an in situ immunostaining assay for transformants, 39 cDNA clones (clone p-16 alpha) encoding mouse liver microsomal testosterone 16 alpha-hydroxylase (cytochrome P-450(16) alpha) were isolated from a cDNA library constructed in the cloning vector pUC-9 with poly(A)+ RNA immunoenriched from total liver polysomes of male 129/J mice. mRNA selected by hybridization with clone p-16 alpha translated the P-450(16) alpha apoprotein in vitro. Total cellular proteins, which were prepared from immunopositive transformant Escherichia coli cells, were conjugated with Sepharose 4B. Antibody purified with the Sepharose 4B conjugate from mixed antiserum to P-450(16) alpha and P-450(15) alpha specifically inhibited testosterone 16 alpha-hydroxylase activity in microsomes. The cDNA insert of one recombinant plasmid (clone P-16 alpha-1) was 1.75 kilobases in size and contained one or more internal restriction sites for HindIII, BamHI, Bgl I, Pst I, Alu I, HinpI, and Rsa I. 32P-labeled clone p-16 alpha-1 hybridized with a single mRNA (2000 bases) that was 10 times more concentrated in liver cells from male 129/J mice than in female mice. This result was consistent with the finding that poly(A)+ RNA from male mice translated 10 times as much P-450(16) alpha in vitro as did the poly(A)+ RNA from females. Thus, the predominant expression of testosterone 16 alpha-hydroxylase in male 129/J mice is regulated pretranslationally, presumably at the transcriptional level of the P-450(16) alpha gene.

Animals↗
PubMed1985

Choice of surgical treatment for multisegmental cervical spondylotic myelopathy.

Three surgical procedures for multisegmental cervical spondylotic myelopathy were evaluated on the basis of a follow-up study (12-157 months) of 95 patients. Twenty-four patients were treated by extensive laminectomy, 50 by anterior interbody fusion by the Cloward and/or Smith-Robinson techniques, and 21 by subtotal spondylectomy and fusion. Results of subtotal spondylectomy were significantly (P less than 0.01) better when compared with those of the other two procedures. It was concluded that spondylosis up to three disc levels should be treated by subtotal spondylectomy and fusion regardless of the canal diameter. When involvement extended four or more levels, extensive laminectomy was recommended.

Cervical Vertebrae↗
PubMed1985

Three functionally distinct helper T-cell clones: the roles for antigen non-specific helper factors in B-cell activation through two different pathways.

We established three functionally distinct purified protein derivative (PPD)-reactive T-cell clones (B11.15, B12.F and D-2). Clone B11.15 could co-operate with DNP-primed B cells to induce anti-DNP IgG plaque-forming cell (PFC) responses only when high amounts of PPD were added to the culture, whereas stimulation of a low amount of DNP-PPD was ineffective (factor-mediated interaction). On the other hand, clone D-2 activated those B cells in a MHC-restricted manner only when DNP-PPD was added to the culture (cognate interaction). B12.F could stimulate B cells with either PPD or DNP-PPD. Antigen non-specific helper factors (lymphokines) responsible for B-cell activation produced by cloned T cells upon stimulation with PPD and antigen-presenting cells were then investigated. Lymphokine activities determined in the present study were IL-2, BCGF I, BCGF II and TRF. BCGF I activity was determined by proliferation-inducing activity on purified B cells in the presence of anti-IgM antibody. BCGF II activity was measured by proliferation-inducing activity on purified B cells in the presence of dextran sulphate. TRF activity was determined on DNP-primed B cells for inducing further differentiation into anti-DNP IgG PFC. BCGF I active molecules were eluted in the fraction at apparent MW of 50,000-70,000 and 8,000-10,000 in gel-permeation column chromatography.

Animals↗
PubMed1985

Purification and physicochemical characterization of murine T cell replacing factor (TRF).

Murine T cell replacing factor (TRF) was purified from a cellfree supernatant of a T cell hybridoma (B151K12) that constitutively produces TRF. Two assay systems for TRF activity were employed: 1) induction of anti-DNP IgG PFC responses in cultures of splenic B cells from DNP-KLH-primed BALB/c mice, and 2) induction of IgM PFC in chronic B cell leukemic cells (BCL1). The purification scheme consisted of ammonium sulfate precipitation, DEAE-cellulose chromatography, Blue-Sepharose chromatography, hydroxylapatite chromatography, gel permeation with fast protein liquid chromatography (FPLC), and disc polyacrylamide gel electrophoresis. Overall, TRF was purified approximately 34,000-fold with a maximum 3.8% recovery of activity, and the specific activity of the purified TRF was approximately 9.6 X 10(4) U/mg. The TRF that is active in these systems is distinct from the other lymphokines such as IL 1, IL 2, BCGFI (now known as BSFp1), and gamma-interferon. The TRF is extremely hydrophobic, with an apparent m.w. of 50,000 to 60,000 on gel permeation chromatography and 18,000 on SDS-PAGE under reducing conditions. Highly purified B151-TRF abrogated the activity by treatment with trypsin but not with RNase. Moreover, it bound to lima bean agglutinin-Sepharose specific for N-acetylgalactosamine residues, indicating that B151-TRF is a glycosylated glycoprotein containing N-acetylgalactosamine residues. The role of N-acetylgalactosamine residues on TRF activity was additionally substantiated by the fact that the addition of appropriate amounts of N-acetylgalactosamine in the assay systems for TRF preferentially induced a profound suppression for TRF-mediated PFC responses.

Adjuvants, Immunologic↗
PubMed1985

BCGFII activity on activated B cells of a purified murine T cell-replacing factor (TRF) from a T cell hybridoma (B151K12).

Experiments were performed to examine a growth-promoting activity on B cells or B leukemic cells of T cell-replacing factor (TRF) produced by a murine T cell hybridoma (B151K12) which constitutively produces TRF. The cellfree supernatant (CFS) from B151K12 cells (B151-CFS) could induce terminal differentiation of pre-activated B cells or in vivo passaged chronic B leukemia cells, BCL1, into immunoglobulin-secreting cells, while it did not exert a nominal lymphokine activity such as BCGFI (now known as BSFpl), IL 2, or gamma-interferon. However, it promoted [3H]thymidine uptake of dextran sulfate (DXS)-stimulated normal B cells and in vivo passaged BCL1 cells, suggesting that it also has BCGFII activity. We tried extensively to purify and to separate the TRF active molecule from the BCGFII active molecule by using many types of purification procedures. The purification scheme consisted of ammonium sulfate precipitation, DEAE-cellulose chromatography, Blue-Sepharose chromatography, hydroxylapatite chromatography, and gel permeation with fast protein liquid chromatography (FPLC). It was revealed that the BCGFII active molecule was hardly separable from the TRF during the entire purification procedure. The TRF as well as BCGFII active materials were glycoprotein with an apparent m.w. of 50 to 60 Kd on gel permeation chromatography and 18 Kd on SDS-PAGE under reducing conditions. The BCGFII active materials were hardly separable from the TRF active one, even after a reverse-phase FPLC, in which both BCGFII and TRF activities were recovered in the fractions eluted at 44 to 48% acetonitrile in 0.1% trifluoroacetic acid (TFA). Furthermore, the absorption of TRF and BCGFII active materials by using BCL1 cells removed not only TRF but also BCGFII activity. Moreover, B cell-specific monoclonal antibody (9T1), which can preferentially block TRF-dependent plaque-forming cell responses, also inhibited the expression of BCGFII activity to BCL1 cells. Taking all of the results together, we conclude that the TRF from B151K12 cells promotes growth of appropriately activated, such as DXS-stimulated normal cells and BCL1 tumor cells. These results suggest that B151-TRF may act on B cells as B cell growth and differentiation factors.

Absorption↗
PubMed1985

[Surgical results of severe acute pancreatitis with special consideration of the severity of pancreatitis].

The surgical treatment of acute pancreatitis remains controversial. Since 1975, we have treated 70 patients with acute pancreatitis. In 36 with severe acute pancreatitis who were not responding adequately to medical intensive treatments, mobilization and drainage of pancreatic exudate from pancreatic bed and retroperitoneal cavity (M-D procedure) were carried out. Thirty-six operative cases were classified into 8 edematous, 7 hemorrhagic, and 21 necrotizing. Edematous pancreatitis showed a tendency of early development of severe symptoms such as enzymatic encephalopathy. Eight of 36 patients who underwent M-D procedure died (22.2 percent), but the rate became 13.8 percent after excluding 3 who died of unrelated causes. Postoperative complications were most frequently encountered in the patients with gallstone pancreatitis. Obstruction of the alimentary tract due to pancreatic or peripancreatic abscess was most serious late complication. Six of the eight were required reoperations and three of them died of further complications which were developed after reoperations. The rate of curability of M-D procedure was 84.8 percent. These data suggest that M-D procedure is highly effective in the treatment of early case of severe acute pancreatitis.

Acute Disease↗
PubMed1984

Mouse liver testosterone 16 alpha-hydroxylase (cytochrome P-450(16) alpha). Purification, regioselectivity, stereospecificity, and immunochemical characterization.

Microsomal testosterone 16 alpha-hydroxylase (cytochrome P-450(16) alpha) was purified from the livers of male 129/J mice based on enzyme activity in the eluates from columns of DEAE Bio-Gel A, hydroxylapatite, and isobutyl-Sepharose 4B. The specific cytochrome P-450 content of the purified P-450(16) alpha fraction was 9.5 nmol/mg of protein. The specific testosterone 16 alpha-hydroxylation activity of the purified P-450(16) alpha fraction was 80 nmol/min/nmol of cytochrome P-450 or 764 nmol/min/mg of protein, and these values were about 40- and 400-fold higher, respectively, than the activity of solubilized microsomes. The purified P-450(16) alpha showed extremely high regioselectivity and stereospecificity for testosterone hydroxylation; more than 90% of the testosterone metabolites formed by the purified P-450(16) alpha fraction was 16 alpha-hydroxytestosterone. The purified anti-P-450(16) alpha antibody exhibited absolute specificity for inhibition of testosterone 16 alpha-hydroxytestosterone was inhibited by the anti-P-450(16) alpha. Anti-P-450(16) alpha inhibited the 16 alpha-hydroxylation activity of intact microsomes prepared from livers of male or female 129/J mice more than 90%, indicating that P-450(16) alpha is the major cytochrome P-450 isozyme catalyzing 16 alpha-hydroxylation activity of testosterone in these microsomal preparations. The purified P-450(16) alpha fraction also possessed high benzphetamine N-demethylation activity relative to the rates found with other xenobiotic substrates tested in this report.

Animals↗
PubMed1984

Kinetic isotope effects on cytochrome P-450-catalyzed oxidation reactions. Evidence for the irreversible formation of an activated oxygen intermediate of cytochrome P-448.

Substitution of deuterium for hydrogen on the alpha-carbon of 7-ethoxycoumarin results in an intrinsic isotope effect of approximately 14 during the cytochrome P-448-catalyzed O-deethylation of this substrate (G. T. Miwa, J. S. Walsh, and A. Y. H. Lu (1984) J. Biol. Chem. 259, 3000-3004). This dramatic decrease in the C-H bond cleavage rate does not, however, alter the rate of 7-ethoxycoumarin disappearance or the rates of NADPH and oxygen consumption indicating that the catalytic turnover of this enzyme is unaffected. Moreover, hydrogen peroxide formation and the concentrations of an oxycytochrome P-448 complex (lambda max = 440 nm) are also unchanged demonstrating that the steady state concentrations of various oxy intermediates of cytochrome P-448 are also unchanged. An inescapable conclusion from these data is that an irreversible step exists between the formation of these intermediates and the oxidation of the substrate. These data are in agreement with the view that an irreversible cleavage of the dioxygen bond precedes substrate oxidation. Moreover, the oxidatively competent form of the cytochrome must then be committed to substrate oxidation. The latter conclusion is substantiated from high performance liquid chromatography studies which demonstrate the formation of a second metabolite, 6-hydroxy-7-ethoxycoumarin, from the deuterated substrate arising from the metabolic switching away from the O-ethyl group to the aromatic ring of this substrate.

Animals↗
PubMed1984

Mouse liver testosterone 15 alpha-hydroxylase (cytochrome P-450(15) alpha). Purification, regioselectivity, stereospecificity, and sex-dependent expression.

Testosterone 15 alpha-hydroxylase (cytochrome P-450(15) alpha) was purified from female 129/J mouse liver microsomes based on its specific activities in the eluates from the columns of octylamino-Sepharose 4B, hydroxylapatite, DEAE-Bio-Gel A, and CM52 chromatography. The 15 alpha-hydroxylation activity was five times higher in female than in male 129/J mouse liver microsomes. The specific cytochrome P-450 content of purified P-450(15) alpha fraction was 14.5 nmol/mg of protein. The Soret peak of the reduced cytochrome P-450-CO complex was 451 nm. The apparent subunit molecular weight of P-450(15) alpha was 48,000, and the protein appeared as only one major band on sodium dodecyl sulfate-polyacrylamide gels. The specific activity of testosterone 15 alpha-hydroxylation reconstituted with the purified P-450(15) alpha was 94 nmol/min/nmol of cytochrome P-450 and 1349 nmol/min/mg of protein, and these were about 65- and 1000-fold higher, respectively, than the activity of solubilized microsomes. The purified P-450(15) alpha exhibited high regioselectivity and stereospecificity for testosterone hydroxylation. More than 95% of the testosterone metabolites formed by the purified P-450(15) alpha was 15 alpha-hydroxytestosterone. Virtually 100% of mouse liver microsomal testosterone 15 alpha-hydroxylation activity can be accounted for by the purified P-450(15) alpha. The P-450(15) alpha fraction was able to catalyze benzphetamine N-demethylation, 7-ethoxycoumarin O-de-ethylation, aniline 4-hydroxylation, benzo(alpha)pyrene 3-hydroxylation, acetanilide 4-hydroxylation, and lauric acid (11 + 12)-hydroxylation at various turnover rates, indicating broad substrate specificity of the P-450(15) alpha for the oxidations of xenobiotics. This is in sharp contrast to high regioselectivity and stereospecificity for testosterone hydroxylation.

Animals↗
PubMed1984

Structure determination of polysaccharides in Aloe saponaria (Hill.) Haw. (Liliaceae).

Neutral polysaccharides that inhibit carrageenin-induced edema in rats were isolated from the nondialysate of the pulp of Aloe saponaria by gel filtration. These were shown to be a linear polymer of a 1,4-linked beta-D-mannopyranose (mol. wt. 15,000) containing 18% acetyl groups (As mannan 1), and a 1,4-linked alpha-D-mannopyranose polymer containing a single branch on the principal chain consisting of D-glucose residues linked at C-2 and C-4 (mol. wt. 66,000), with 10% acetyl groups (As mannan 2). As mannan 1 inhibited carrageenin-induced hind paw edema at 50 mg/kg ip in rats; As mannan 2 was not tested for pharmacological activity. A crude preparation of both As mannans was effective when given intraperitoneally, but was ineffective when given orally.

Aloe↗
PubMed1984

Validity of various function tests performed in Japan as a screening test for vibration syndrome.

In Japan, various function tests on the upper extremities are widely performed for the early diagnosis of vibration syndrome. The validity of these function tests was investigated by discriminant function analysis. The correct classification rate of the 120 workers exposed to vibration and the 40 control workers was 83.1%, when including the exposure-to-cold test. Therefore, the function tests were confirmed to be of diagnostic significance as a screening test for vibration syndrome. However, the rate is not sufficient to make a diagnosis only with the function tests. When not including the exposure-to-cold test, the rate was reduced by 10% because insidious functional disorders were missed in the early stage. For the early diagnosis of vibration syndrome, the exposure-to-cold test should be included in the function tests. Furthermore, to enhance the validity of the diagnosis, the results of the function tests should be evaluated together with subjective symptoms and the findings on bones and joints, the central nervous system and the endocrine system.

Adult↗
PubMed1984

Results of mobilization and drainage of the pancreas for acute pancreatitis.

The surgical treatment of acute pancreatitis remains controversial. Since 1969, we treated 60 patients with acute pancreatitis. In 34 with severe acute pancreatitis who were not responding adequately to intensive medical care, surgical intervention was made by mobilization of the pancreas from retroperitoneal tissue and drainage of the pancreatic bed (M-D procedure). Thirty-four operative cases were classified into 7 edematous, 7 hemorrhagic, and 20 necrotizing. Macroscopic findings of the pancreas did not correlate either to the severity of the acute pancreatitis or to the mortality rate. Eight of 34 who underwent M-D procedure died (23 per cent), but the rate became 14.7 per cent after excluding 3 who died of unrelated causes. These data suggest that the M-D procedure is highly effective in the treatment of early cases of severe acute pancreatitis. There was an associated marked reduction in the mortality rate with the prophylactic use of broad spectrum antibiotics. With M-D procedure, there was a low incidence of late sequelae of pancreatic or peripancreatic abscess.

Acute Disease↗