Effect of ionic strength on production of cAMP- and Ca2+-independent protein kinase from rat liver plasma membrane.

Production of cAMP- and Ca2+-independent protein kinase was stimulated when rat liver plasma membrane was incubated with increasing concentrations of NaCl. This protein kinase release was diminished by addition of protease inhibitor. The molecular mass of this enzyme was approx. 50 kDa and a high concentration of Mg2+ was required for whole histone phosphorylation. These properties are similar to those of the protease-activated form of protein kinase C. The NaCl effect could be replaced by other salts such as LiCl and NaHCO3. These results suggest that membrane-bound protein kinase C is activated by limited proteolysis corresponding to an increase in ionic strength.

A reverse flow-metabolism mismatch pattern on PET is related to multivessel disease in patients with acute myocardial infarction.

UNLABELLED: Hypoperfused myocardium with increased uptake of 18F-fluorodeoxyglucose (FDG) is considered to be ischemic but viable myocardium. However, the significance of a more severe defect of FDG than of 13N ammonia (NH3) (i.e., reverse flow-metabolism mismatch) is not well understood. METHODS: To study a reverse flow-metabolism mismatch pattern, PET with NH3 and FDG under glucose loading was performed in 35 patients within 2 wk after onset of first acute myocardial infarction (AMI) and in 29 patients with old myocardial infarction (OMI). The left ventricle was divided into nine segments on a bull's eye polar map, and the mean counts of NH3 (%NH3) and FDG (%FDG) were compared for the segment with the least %NH3. RESULTS: Ten patients in the AMI group demonstrated a marked reverse flow-metabolism mismatch pattern (greater than 10% difference between %NH3 and %FDG), whereas only 2 patients in the OMI group demonstrated the mismatch pattern (P < 0.05). Sixteen patients with AMI demonstrated %FDG > %NH3 (group 1), and 19 patients with AMI demonstrated %FDG < %NH3 (group 2). There were no significant differences in age, sex, location of infarction, diameter of stenosis of infarct-related artery or left ventricular ejection fraction between groups 1 and 2. Eleven patients in group 2 and only 3 in group 1 had multivessel disease (P < 0.02). There was no significant relationship between the number of diseased vessels and the flow-metabolism pattern in patients with OMI. CONCLUSION: The finding of a reverse flow-metabolism mismatch on PET in the subacute phase of myocardial infarction was closely related to multivessel disease.

Effects of preinfarction angina on myocardial injury in patients with acute myocardial infarction: a study with resting 123I-BMIPP and 201T1 myocardial SPECT.

METHODS: Recent studies have suggested that patients with preinfarction angina have smaller infarcts and a better in-hospital outcome than those without angina. The mechanisms responsible for limitation of infarct size in the presence of preinfarction angina are unclear. We examined the effects of preinfarction angina on myocardial injury in patients with the first acute myocardial infarction with resting 123I-15-(p-iodophenyl)-3-(R,S)-methylpentadecanoic acid (BMIPP) 201TI myocardial scanning performed within 1 mo of infarction. RESULTS: Of 136 patients tested, 48 (35%) had preinfarction angina within 72 h before infarction, whereas 88 (65%) did not. BMIPP and 201TI defects were scored in 9 segments of the left ventricle (0 = normal, 1 = mild defect, 2 = moderate defect, 3 = severe defect, and 4 = no uptake). The total defect score was defined as the sum of the defect scores. There was no significant difference in percentage diameters of stenoses of infarct-related arteries, collateral circulation, total defect scores for BMIPP, or 201TI between the groups with and without preinfarction angina. However, the ratio of total defect score for 201TI to that for BMIPP was significantly smaller for patients with than for those without preinfarction angina (0.64 +/- 0.21 versus 0.74 +/- 0.25, respectively; P = 0.007). CONCLUSION: Preinfarction angina did not affect the areas at risk in acute myocardial infarction, as shown by BMIPP defect, but decreased necrotic myocardium in the areas at risk, as shown by 201TI defect, and increased metabolically damaged but viable myocardium, as shown by BMIPP and 201TI mismatch through unidentified mechanisms other than collateral circulation (e.g., ischemic preconditioning).

Ionic strength-dependent proteolytic activation of protein kinase C by trypsin-like protease.

Protein kinase C, reversibly bound to rat liver plasma membrane through Ca2+, was activated by endogenous trypsin-like protease in an ionic strength-dependent manner. In an attempt to understand the reaction mechanism, the EGTA-extracted protein kinase C and the trypsin-like protease (Tanaka, K. et al. (1986) J. Biol. Chem. 261, 2610-2615) were separately purified from plasma membrane. In the reaction system using these purified enzymes, increasing the ionic strength with NaCl (140-210 mM) effectively enhanced the proteolytic activation of the protein kinase C in the presence of Ca2+ and phospholipid. These results suggest that ionic strength is an important factor for the proteolytic activation of membrane-bound rat liver protein kinase C.

Effects of sodium valproate on various intestinal motor functions after ileal J pouch-anal anastomosis.

BACKGROUND: We investigate the effects of sodium valproate, a gamma-aminobutyric acid transaminase inhibitor, on the clinico-physiologic state after ileoanal anastomosis. METHODS: Twelve patients (five with ulcerative colitis and seven with familial adenomatous polyposis coli) were studied in a placebo-controlled trial. RESULTS: Placebo had no effect, but valproate increased the resting anal pressure by 22.4% and the maximum tolerable pouch volume by 20.7%. It also prolonged pouch emptying by 47.3%, with an associated improvement in stool frequency and incontinence. These results suggested that valproate improved both reservoir continence and sphincter continence. CONCLUSIONS: Only minimal side effects were noted, so the administration of sodium valproate appears to be a safe and useful adjunct for patients with ileoanal anastomosis in the early postoperative period.

Modified anoabdominal rectal resection and colonic J-pouch anal anastomosis for lower rectal carcinoma: preliminary report.

BACKGROUND: There has recently been increasing interest in coloanal reconstruction after proctectomy for low rectal carcinoma. We describe here our pilot experience with seven patients undergoing modified anoabdominal resection of the rectum and a colonic J-pouch anal anastomosis. METHODS: The procedure varied according to the extent of internal anal sphincter (IAS) resection (type a, partial resection of the upper IAS; type b, circumferential resection of the upper IAS; type c, partial preservation of the lower IAS; and type d, total resection of the IAS). RESULTS: None of the patients had incontinence, but preservation of the lower half of the IAS (types a and b) showed functional superiority over more extensive IAS resection (types c and d). Only patients who underwent types c and d resection needed medications to reduce stool frequency. CONCLUSIONS: Our results suggest that the lower half of the IAS has a more important role than the upper half in the control of defecation. Total resection of the IAS did not sacrifice continence, but preservation of at least the lower IAS resulted in a better quality of life. Careful patient selection is needed when considering the use of these procedures for tumors in the lowest part of the rectum.

Lipocortin-present perforating and lipocortin-absent nonperforating Crohn's disease.

To investigate whether Crohn's disease has two different clinical forms, a relatively aggressive perforating type and a more indolent nonperforating type, we compared the concentrations of lipocortin and prostaglandin E2 (PGE2) in the inflamed mucosa of 12 patients with strictly controlled Crohn's disease with those found in histologically normal mucosa of control subjects. The inflamed mucosa obtained from eight patients with nonperforating Crohn's disease did not react with antilipocortin antibody on immune blot analysis. In contrast, the inflamed mucosa from four patients with perforating Crohn's disease, as well as that obtained from histologically normal controls, contained lipocortins. In addition, higher concentrations of intramucosal ileal and colonic PGE2 were found in patients with nonperforating Crohn's disease (902 +/- 454 pg/wet weight [WW] mg and 1,268 +/- 567 pg/WW mg, respectively) compared with normal controls (90.2 +/- 43.1 pg/WW mg and 173 +/- 76.5 pg/WW mg, respectively) (p less than 0.01). The difference in intramucosal ileal and colonic PGE2 levels between patients with perforating Crohn's disease (109.6 +/- 16.7 pg/WW mg and 252 +/- 34.4 pg/WW mg, respectively) and normal controls was not significant. These findings indicate that there may be two distinct patterns of Crohn's disease that differ in the amount of lipocortin present in the intestinal mucosa.

Hemodynamics in the prone jackknife position during surgery.

We examined the hemodynamic changes occurring with prone jackknife positioning during colorectal surgery. The operative procedure was restorative proctocolectomy with ileal J-pouch anal anastomosis in five patients with adenomatosis coli and six patients with ulcerative colitis and anoabdominal resection of the rectum with colonic J-pouch anal anastomosis in eight patients with rectal cancer. Nineteen patients (10 men and 9 women aged 41 +/- 19 years) were monitored with arterial and Swan-Ganz catheters during positioning. Measurements were obtained in the supine and prone positions (1 minute, 3 minutes), and the jackknife position (1, 3, 5, and 10 minutes), as well as before and after adoption of the Lloyd-Davies position (1, 3, 5, and 10 minutes). Turning the patient from the supine position to the prone position resulted in a significant decrease in the cardiac index (CI). However, following head-down rotation, the CI increased and returned to the value seen in the supine position (p less than 0.05). Heart rate (HR) slowed and mean arterial pressure (MAP) increased in the prone jackknife position. We concluded that the extent of the changes in cardiac function presented no serious problems.

Gastric volvulus complicating myotonic dystrophy.

Myotonic dystrophy is an autosomal inherited disorder of both striated and smooth muscle, and is considered to be a rare cause of gastrointestinal dilatation and abnormal peristalsis. We report on a patient with myotonic dystrophy complicated by gastric volvulus. A 57-year-old female with myotonic dystrophy suddenly developed abdominal pain, nausea and vomiting. X-ray examinations revealed gastric dilatation and pyloroantral obstruction, consistent with acute gastric volvulus. The patient underwent successful emergency gastrectomy. Gastric volvulus is often an unrecognized surgical emergency, but its clinical and radiographic features are so characteristic that accurate diagnosis is possible if the condition is kept in mind. Thus, the clinician should consider the possibility of gastric volvulus when evaluating gastrointestinal complaints in patients with myotonic dystrophy.

Problems in the initial diagnosis of renal infarction.

We retrospectively analyzed 20 cases of renal infarction to identify the problems in tentatively diagnosing renal infarction. The subjects consisted of 12 outpatients and 8 inpatients whose diagnosis was confirmed by renal scintigram and/or contrast computed tomography. Renal infarction was tentatively diagnosed in only 4 of the 12 outpatients. Causes of hospitalization were cerebral emboli in 5 cases, peripheral emboli in the extremities in 2 cases and one case involved percutaneous transmitral commissurotomy. On initial urinalysis, 11 cases (55%) showed less than 2+ hematuria using dipsticks to test for occult blood. The mean lactic dehydrogenase value was as high as 2,096 IU while the mean aspartate aminotransferase and mean alanine aminotransferase were 83.1 IU and 78.6 IU. Abdominal ultrasonography revealed abnormalities in only one of 18 cases. In conclusion, since only a moderate degree of hematuria was seen in about half the cases and it was difficult to detect renal abnormalities by ultrasonography, a tentative diagnosis of renal infarction may be difficult in some cases.

Problems in pharmacological evaluation of patients with paroxysmal atrial fibrillation: clinical analysis of more than 100 consecutive patients.

To address the problems of pharmacological evaluation in paroxysmal atrial fibrillation (PAf), we interviewed 108 consecutive patients with documented PAf regarding symptoms, frequency and trigger factors of PAf and analyzed the 24-hour ambulatory electrocardiographic monitoring (Holter monitoring) records in relation to symptoms. Twenty-nine patients were totally asymptomatic, while 79 patients were symptomatic of which 49 patients had obvious trigger factors. PAf was documented by Holter monitoring in 22 of 79 symptomatic patients. On analysis of PAf-documented 25 Holter monitoring records, the patients checked event marks as PAf in only 20 of 155 PAf episodes. Six episodes of 26 event marks that patients thought to be PAf proved to be premature atrial or ventricular contractions. Nine patients in whom PAf persisted for more than 24 hours became asymptomatic. Patients suitable for pharmacological evaluation constituted about one-fifth of the PAf patients in our consecutive study. Even with the selection of these patients, pharmacological evaluation based on symptoms is difficult because disappearance of PAf may be associated with persistent atrial fibrillation.

Influence of inoculation site of combined oil-adjuvanted vaccine on the antibody response in chickens.

Inactivated oil-adjuvanted vaccines for ND, IB, and IC serotypes A and C (OILVAX NB2AC) have been marketed from 1993. In the outdoors, various inoculation sites have been used in chickens because to make the inoculation procedure easier. We examined whether differences would be obtained in the antibody response according to the inoculation sites; the subcutaneous inoculation into the back of the neck for OILVAX use, and the thigh, lower thigh, breast and shoulder muscle for the possible application of the inoculation outdoors. The clear order was not found in the NDV-HI titer, IBV-SN titer against Nerima or TM-86 among the inoculation sites used during the examination period. The IC serotypes A and C HI titers did vary among the inoculation sites; the subcutaneous inoculation produced the highest antibody titer, and high antibody titers were observed in the order of lower thigh muscle > or = thigh muscle > breast muscle > or = shoulder muscle inoculation.

Intravenous myocardial contrast echocardiography predicts regional and global left ventricular remodelling after acute myocardial infarction: comparison with low dose dobutamine stress echocardiography.

OBJECTIVE: To assess the role of intravenous myocardial contrast echocardiography (MCE) in predicting functional recovery and regional or global left ventricular (LV) remodelling after acute myocardial infarction (AMI) compared with low dose dobutamine stress echocardiography (LDSE). METHODS: 21 patients with anterior AMI and successful primary angioplasty underwent MCE and LDSE during the subacute stage (2-4 weeks after AMI). Myocardial perfusion and contractile reserve were assessed in each segment (12 segment model) with MCE and LDSE. The 118 dyssynergic segments in the subacute stage were classified as recovered, unchanged, or remodelled according to wall motion at six months' follow up. Percentage increase in LV end diastolic volume (%DeltaEDV) was also calculated. RESULTS: The presence of perfusion was less accurate than the presence of contractile reserve in predicting regional recovery (55% v 81%, p < 0.0001). However, the absence of perfusion was more accurate than the absence of contractile reserve in predicting regional remodelling (83% v 48%, p < 0.0001). The number of segments without perfusion was an independent predictor of %DeltaEDV, whereas the number of segments without contractile reserve was not. The area under the receiver operating characteristic curve showed that the number of segments without perfusion predicted substantial LV dilatation (%DeltaEDV > 20%) more accurately than did the number of segments without contractile reserve (0.88 v 0.72). CONCLUSION: In successfully revascularised patients with AMI, myocardial perfusion assessed by MCE is predictive of regional and global LV remodelling rather than of functional recovery, whereas contractile reserve assessed by LDSE is predictive of functional recovery rather than of LV remodelling.

Insulin-stimulated serine kinase in Xenopus oocyte plasma membrane.

Insulin-stimulated protein kinase activities detected in Xenopus oocyte membrane were examined. The plasma membrane proteins solubilized in a buffer containing Triton X-100 were immunoprecipitated with anti-phosphotyrosine antibodies and adsorbed materials were eluted with a buffer containing p-nitrophenyl phosphate. The eluate contained protein serine kinase activity toward H1 histone which was increased 2-3 fold by insulin. Protein tyrosine kinase activity was also exhibited in Xenopus oocyte membrane and the close parallel to serine kinase activity was observed in response to insulin. These results suggest that insulin-stimulated serine kinase is activated through the phosphorylation by protein tyrosine kinase.

Intraoperative colonoscopy.

From 1987 to 1991, we performed intraoperative colonoscopy on 66 patients as a result of the following indications: the air leakage test (53%), detection of a previous malignant polypectomy site (20%), inability to complete preoperative colonoscopy (17%), detection of the source of intestinal bleeding (4%), and detection of impalpable colonic lesions (4%). Intraoperative colonoscopy was successful in 61/66 patients and provided information that altered the planned operation in 10 of 61 completely examined patients. The air leakage test proved useful in detecting subclinical anastomotic leaks. The indications for intraoperative colonoscopy have been expanded, and this procedure is often useful when one is attempting to decide on the appropriate surgical strategy for patients.

Variable activity of protein tyrosine kinase in apparently normal thyroid glands.

We examined protein tyrosine kinase (PTK) activity in apparently normal thyroid tissue (n = 22) obtained from patients undergoing thyroid surgery for papillary thyroid cancer or benign disease. The PTK activity in apparently normal thyroid tissue from patients with papillary thyroid cancer progressively was elevated in the cytosolic fraction compared with that from patients without cancer (p < 0.05). The cytosolic proportion of PTK activity was also significantly increased in the normal thyroid tissue of patients with cancer (62% vs. 51%, p < 0.05). These findings suggest that alterations in PTK activity may indicate apparently normal thyroid tissue at increased risk of developing malignancy.

Stapled anastomosis for trans-sphincteric resection of the rectum.

We have found that in all patients who have undergone this procedure, normal defecation and complete continence are maintained. The surgical wound heals well and functional results have been uniformly good, even in instances in which only 1 centimeter of the rectum remains. This procedure may be useful for patients who are carefully diagnosed by preoperative transrectal ultrasonography in which the lower edge of the rectal tumor is within 8 centimeters of the anal verge, the tumors do not extend into the muscularis propria and there is no lymph node metastasis. Use of the CEEA stapling device can shorten the time of operation and is particularly beneficial for elderly patients and those who are at poor risk.

Altered protein tyrosine kinase levels in human colon carcinoma.

To further understand the molecular mechanisms and the biological indicators of colonic tumorigenesis, the authors examined tyrosine kinase activity in the cytosol and in the particulate fraction of the homogenates of specimens from 20 human colonic carcinomas and compared them with the adjacent normal mucosal tissues. Total protein tyrosine kinase activity could be precisely detected using miniphosphocellulose column purification and a synthetic peptide, Glu-asparagine (Asp)-alanine (Ala)-Glu-tyrosine (Tyr)-Ala-Ala-arginine (Arg)-Arg-Arg-glycine (Gly) (E11-G1), as an artificial substrate. Tyrosine kinase activity of colonic carcinoma and normal mucosa was reduced in the cytosol fraction whereas activity in the particulate fraction was elevated with respect to protein concentration. The average specific activity ratios were 1.95 +/- 0.27 (normal cytosolic/carcinoma cytosolic) and 0.57 +/- 0.01 (normal particulate/carcinoma particulate) for tyrosine kinase activity. Cellular distribution (% cytosol) of tyrosine kinase activity in normal mucosa and in carcinoma varied from 21.0% to 91.2% and from 7.0% to 61.4%, respectively. In nearly all cases the percentage of cytosolic tyrosine kinase activity in carcinoma tissues was lower than in normal tissues. There was no difference due to histologic type or the presence of adenomatous components. A significant decrease of cytosolic tyrosine kinases was correlated with Dukes' Stage A. With advancing Dukes' stage, the average specific activity ratios (normal cytosol/carcinoma cytosol) were decreased. This study indicates that colonic carcinogenesis might be associated with alterations in cellular levels of tyrosine kinase activity and that the average specific activity ratio (normal cytosol/carcinoma cytosol) had a possible correlation with colonic tumor growth.