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Using a human-mouse monochromosomal hybrid, BG15-6, that contains an intact human chromosome 5, we isolated four monoclonal antibodies, 2A10, 3H9, 5G9, and 6G12, as chromosome marker antibodies recognizing cell surface antigens specific for human chromosome 5. The binding patterns of these antibodies to BG15 subclones containing fragments of human chromosome 5 indicated that 2A10, 3H9, and 6G12 recognized the antigens produced by genes located on 5pter-q22, and that 5G9 recognized the antigen produced by a gene located on 5q23. Cells containing human chromosome 5 were very effectively sorted in a fluorescence-activated cell sorter (FACS) using monoclonal antibody 6G12. This method for sorting cells containing human chromosome 5 or an appropriate fragment of this chromosome from among human-rodent hybrid cells should be very useful in studies on gene expression, gene cloning and gene mapping.
We propose a chaos-synchronization scheme using incoherent feedback to the pumping power in two microchip lasers. The population inversion of the slave laser is controlled for synchronization by using the detected signals of the peak heights of chaotic pulse intensities in the two lasers. Matching of the optical frequencies between the two lasers (i.e., injection locking) is not required for synchronization using this method. We numerically demonstrate the incoherent feedback method and investigate synchronization regions against parameter mismatching between the two lasers. Synchronization is maintained within a mismatching of 1% for all laser parameters, which implies that the difficulty in reproducing the synchronized laser pulses is very useful for applications of secure optical communications.
An ATP bioluminescence assay as a rapid reference method for fluconazole (FLCZ) susceptibility testing of dermatophytes, as well as yeasts, was developed and evaluated by comparing it with viability, turbidity and fungal protein content-based conventional methods. FLCZ susceptibility results obtained with strains of Candida albicans and dermatophytes by the bioluminescence method in high-resolution medium were well correlated with those obtained by conventional methods currently used in clinical microbiology laboratories or reported previously, including a broth dilution method by the National Committee for Clinical Laboratory Standards (NCCLS). Thus, ATP bioluminescence assay can be used to monitor fungal growth in liquid culture media. The procedure has considerable potential for the rapid testing of FLCZ susceptibility of dermatophytes and other fungi.
In the present study, a sequential observation of the gastric lesions or ulcers in rats weighing from 100 g was made with an endoscope. It was found that bleeding, edema, hyperemia, fur of ulcer and convergences of the mucosal folds in the stomach could be observed in rats with a SES-1717S (phi = 1.7 mm) or SES-2217S (phi = 2.2 mm) endoscope. Furthermore, we have devised a method which we named the "sohmen method" for measuring ulcer-sizes with an endoscope. The ulcer-size was compared with 5 mm lengths of sohmen (Japanease vermicelli) which were inserted through the sheath of the endoscope into the stomach. Therefore, the ulcer-size could be accurately measured by this method. By this observation method, it is possible to observe repeatedly over a long term the changes in gastric lesions or ulcers, to measure changes in ulcer-sizes and to evaluate the relapse or recurrence of ulcers in the same rats.
We reported the technique of pathological diagnosis for minute lung cancer lesion of early stage during operation. Target lesion for our technique may be less than 1 cm in size and not be detected by palpation. Our technique may superior to usual method of frozen section of tissue specimen without fixation. At first, resected lung specimen was fixed by injection of 20% formalin solution subsequent warming by microwave oven for 40 seconds. Almost all lesions may be detected in proper thickness of specimen. Specimen was washed out and steeped in the 20% of sucrose solution for 30 to 60 seconds in order to avoid crystallization of water that was contained within lung specimen. Then thin sliced section of specimen was made by Cryostat in usual method. The minute cancer lesion was able to detect more easy than usual method of making frozen section of specimen. Formalin fixed and sucrose steeped frozen section may be able to evaluate the surgical margin from cancer exactly than the section made by non-fixed specimen.
By affinity chromatography with Sepharose coupled to 2'-deoxy-1-beta-D-ribofuranosyl-N4-dodecanoylcytosine, deoxycytidine kinase and cytidine deaminase were purified 1,950- and 2,240-fold, respectively, from Ehrlich carcinoma cells, and their enzyme activities for several deoxycytidine analogs were investigated.
Purpose of the present study was to evaluate the myopathy risk using a urethane infusion method following oral administration of five kinds of commercial HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme A) reductase inhibitors (HCRIs), (pravastatin (PV), simvastatin (SV), cerivastatin (CeV), atorvastatin (AV), and fluvastatin (FV)) alone or with coadministration of bezafibrate (BF). The solubility of HCRIs in various solvents was determined as a criterion of the physicochemical property. The plasma creatine phosphokinase (CPK) level as a marker of myopathy in normal rats was screened under urethane infusion after oral administration of HCRI alone or with BF coadministration. Also, renal tissue specimens were prepared and the myoglobin remaining in the tissue was visualized by the labeled avidin-biotin technique. The plasma CPK level in normal rats under urethane infusion following oral administration of five kinds of HCRI increased as the dose of HCRI increased, and coadministration of BF further increased the CPK level for each drug. The risk of myopathy evaluated from the CPK level was ranked as follows: CeV>FV>AV>SV>PV. Myoglobin deposition was observed in the cast of proximal tubules, cytoplasm of distal tubules and collecting ducts of rat kidney extracted from rats treated with HCRIs under urethane infusion. Histopathological findings showed that the extent of myoglobin deposition increased on coadministration of BF with each drug. The correlation was found for myopathy risk evaluated by CPK level using the urethane infusion method and drug lipophilicity, ie., the water/n-octanol partition coefficient except for the case of SV. Histopathological findings for the kidney following HCRI treatment also reflected the CPK level in rats under urethane infusion.
We performed this study to develop a reliable method for inducing colitis in rats using 2,4,6-trinitrobenzene sulfonic acid (TNBS) to reduce the variation in ulcer size. A pair of ring forceps was used to clamp the colon and 0.1 M TNBS in ethanol was injected into the luminal side of the clamped portion. This method resulted in a small coefficient of variation of the ulcer index. A significant linearity was observed by plotting ulcer size against days after ulcer induction in both logarithm scales. These findings show that this technique is simple and reliable and that ulcers heal linearly.
BACKGROUND: The effects of angiotensin antagonists on coronary circulation in type 2 diabetes are unclear. We aimed to assess whether 4 weeks of treatment with angiotensin-converting enzyme inhibitor or angiotensin II type 1 receptor antagonist improves coronary flow velocity reserve (CFVR) in patients with type 2 diabetes. METHODS: Twenty-four asymptomatic patients with type 2 diabetes were randomly assigned to temocapril (2 mg/d) or candesartan (8 mg/d). Coronary flow velocity reserve, calculated as the ratio of adenosine-induced hyperemic to basal coronary flow velocity, was measured with transthoracic Doppler echocardiography. Coronary flow velocity reserve measurement and venous blood sampling were performed before and after 4 weeks of treatment. We also obtained CFVR and venous blood data in the 8 healthy controls. RESULTS: Coronary flow velocity reserve was significantly lower in patients than controls (temocapril group 2.74 +/- 0.28, candesartan group 2.65 +/- 0.30, controls 3.53 +/- 0.23, P < .0001 for both, respectively). Blood pressure was reduced in both diabetic groups (n = 12 each) similarly 4 weeks after treatment. There were no significant differences between the 2 groups in venous blood data before or after treatment. However, CFVR increased significantly in the temocapril group (2.74 +/- 0.28 to 3.31 +/- 0.36, P < .0001), but not in the candesartan group (2.65 +/- 0.30 to 2.71 +/- 0.43, P = ns). CONCLUSIONS: Coronary flow velocity reserve in patients with type 2 diabetes improved after treatment with temocapril but not with candesartan, suggesting that angiotensin-converting enzyme inhibitor, but not angiotensin II type 1 receptor antagonist, might have beneficial effects on coronary microangiopathy associated with type 2 diabetes.
The ribose-modified chromophoric and fluorescent analog of ATP, 2',3'-O-(2,4,6-trinitrocyclohexadienylidene)-ATP (TNP-ATP) (Hiratsuka, T., and Uchida, K. (1973) Biochim. Biophys. Acta 320, 635-647 and Hiratsuka, T. (1976) Biochim. Biophys. Acta 453, 293-297) has been widely used as an ATP analog for various ATPases. Although the corresponding analog of GTP,2',3'-O-(2,4,6-trinitrocyclohexadienylidene)-GTP (TNP-GTP) should be useful for the study of various GTP-requiring enzymes, it is difficult to prepare TNP-GTP by the conventional method. In the present study, we succeeded in the synthesis of TNP-GTP with the use of an alternative method. The analogs of GDP, GMP, and guanyl-5'-yl imidodiphosphate (Gpp(NH)p) were also synthesized. Visible absorption and fluorescent properties of TNP-GTP, TNP-GDP, TNP-GMP, and TNP-Gpp(NH)p were quite similar to those of TNP-ATP. TNP-GTP was found to be able to replace GTP as an inhibitor for bovine liver glutamate dehydrogenase. The enzyme was inhibited by TNP-GTP to a maximum extent of 54% at saturating concentrations of the analog with a KI of 2.7 microM. TNP-Gpp(NH)p and other ribose-modified fluorescent analogs of GTP,3'-O-anthraniloyl-GTP and 3'-O-(N-methylanthraniloyl)-GTP (Hiratsuka, T. (1983) Biochim. Biophys. Acta 742, 496-508), also inhibited the enzymatic activity. Binding of TNP-GTP to the enzyme was characterized by a 5.6-fold enhancement in analog fluorescence. In the presence of NADH, the limiting fluorescence enhancement of the bound analog decreased to 2.7-fold. As determined by fluorometric titration, the maximum number of TNP-GTP binding sites on the enzyme was 1.9 mol/mol of subunit with a KD of 0.66 microM in the absence of NADH and 2.2 mol/mol of subunit with two KD values of 0.11 and 0.71 microM in the presence of NADH. These observations suggest that NADH binding increases the affinity of only 1 mol of the 2 mol of TNP-GTP bound to the enzyme. These spectroscopic and biological properties of TNP-GTP should make this analog useful as a chromophoric and fluorescent probe for studies not only of glutamate dehydrogenase but also of various GTP-requiring enzymes, which have a high specificity for the base moiety of GTP.
Transcatheter arterial embolization (TAE) was performed in 97 patients with hepatocellular carcinoma. Ethiodol (iodized oil) containing an anticancer drug was infused via the hepatic artery followed by Gelfoam particles. The Ethiodol emulsion was selectively retained in the tumor vessels and also remained in the small daughter nodules that could not be detected by angiography or computed tomography (CT) prior to TAE. Ethiodol remained in the tumor for more than 1 year. Following TAE, main tumors and small daughter nodules appeared as areas of markedly high density on CT. In most patients there was a reduction in the tumor size following TAE, and serum alpha-fetoprotein levels were reduced in all patients whose initial levels had exceeded 400 ng/ml. This method is considered to be effective not only for treatment of hepatic tumor but also useful for evaluation of post-TAE changes in the tumor and diagnosis of small daughter nodules, due to the long-term accumulation of Ethiodol in tumor vessels.
The purpose of this study is to analyze our treatment experience on patients with ascending aortic aneurysms, with aortic regurgitation. From January 1974 to December 1995, 49 patients underwent replacement of the aortic valve and ascending aorta with a composite graft, in which primary operation cases were 44 and reoperation ones were 5. The Bentall technique was used in 20 patients, the button technique in 11, the interposition graft technique in 11, and a combination of the interposition graft and button technique in 7. All but one reoperation cases underwent the interposition graft technique. Hospital mortality was 30% for the Bentall technique, and 9.1% for the button technique and 9.1% for the interposition graft technique; there was no hospital mortality in the combination of the interposition graft and button technique. Hospital mortality of interposition graft technique in primary operation cases was 9.1%, and that in reoperation cases was 0%. Hospital mortality in patients underwent from 1974 to 1985 was 30.8%, 27.8% from 1986 to 1991, and 0% from 1992 to 1995. Five late deaths occurred in the Bentall group (35.7%) and one late death in the button technique (9.1%). Not late deaths in the other groups have occurred. In summary, operative mortality in Bentall technique group was higher than that of the other groups. Operative results were improved by the change of operative methods. The interposition graft technique is preferable for patients undergoing reoperation or when tension on the ostial anastomoses may occur. The button technique is best for patients with aortic dissection or inflammation involving the coronary ostia.
This study evaluated the effects of the temporary occlusion and aspiration device GuardWire on percutaneous coronary intervention for acute myocardial infarction. This device brought about a significant reduction in the no-reflow phenomenon and improvement in ST resolution compared with conventional percutaneous coronary intervention.
Magnetic resonance cholangiopancreatography (MRCP) has become widely used due to the rapid advances made as a non-invasive diagnostic method for the pancreatobiliary ducts. We report a case with annular pancreas in an adult in whom the pancreatic duct in the annular portion could be clearly visualized by magnetic resonance cholangiopancreatography, which was very useful for the definite diagnosis as annular pancreas.
Troglitazone increased cardiac output and stroke volume, as a result of decreased peripheral resistance, in diabetic patients with normal cardiac function. The cardiovascular effects of troglitazone in patients with heart failure are unknown. The aim of the study was to evaluate the cardiovascular effects of troglitazone in patients with heart failure. Blood pressure and echocardiographic findings were evaluated before and 1, 2, 3 and 4 hours after a single dose of troglitazone (400 mg) or placebo, in eight type II diabetic patients with congestive heart failure. The plasma catecholamines and coefficient of variance of RR intervals (CVRR) were also measured. Neither heart rate nor blood pressure changed after the administration of troglitazone. Left ventricular (LV) end-diastolic dimension did not change either, however, the LV end-systolic dimension significantly decreased compared with its baseline value and with that of the placebo group. On the other hand, the % fractional shortening and the E/A ratio increased significantly after troglitazone. The LV end-diastolic volume did not change, whereas the LV end-systolic volume significantly decreased. The stroke volume and the LV ejection fraction significantly increased compared with its baseline value and with that of the placebo group. The peripheral vascular resistance did not change after the administration of troglitazone, whereas plasma catecholamines significantly decreased, and CVRR remained unchanged in both groups. These hemodynamic changes suggest that a single oral dose of troglitazone induced inotropy without activation of the sympathetic nervous system.
Extrageniculate projections to the visual cortex were examined in the macaque monkeys by the horseradish peroxidase (HRP) method. Extrageniculate neurons sending fibers to the visual cortex were found in the lateral and inferior pulvinar nuclei, paracentral thalamic nucleus, claustrum, basal nucleus of Meynert, lateral part of the basal amygdaloid nucleus, lateral hypothalamus, locus coeruleus, and dorsomedial and midline regions of the pontine tegmentum.
PURPOSE: The purpose of the study was to develop a method for the quantitative prediction of the bitterness suppression of elemental diets by various flavors and to predict the optimum composition of such elemental diets for oral administration using a multichannel taste sensor. METHODS: We examined the effects of varying the volume of water used for dilution and of adding varying quantities of five flavors (pineapple, apple, milky coffee, powdered green tea, and banana) on the bitterness of the elemental diet, Aminoreban EN. Gustatory sensation tests with human volunteers (n = 9) and measurements using the artificial taste sensor were performed on 50 g Aminoreban EN dissolved in various volumes (140), 180, 220, 260, 300, 420, 660, 1140, and 2100 ml) of water, and on 50 g Aminoreban EN dissolved in 180 ml of water with the addition of 3-9 g of various flavors for taste masking. RESULTS: In gustatory sensation tests, the relationship between the logarithmic values of the volumes of water used for dilution and the bitterness intensity scores awarded by the volunteers proved to be linear. The addition of flavors also reduced the bitterness of elemental diets in gustatory sensation tests; the magnitude of this effect was, in decreasing order, apple, pineapple, milky coffee, powdered green tea, and banana. With the artificial taste sensor, large changes of membrane potential in channel 1, caused by adsorption (CPA values, corresponding to a bitter aftertaste), were observed for Aminoreban EN but not for any of the flavors. There was a good correlation between the CPA values in channel 1 and the results of the human gustatory tests, indicating that the taste sensor is capable of evaluating not only the bitterness of Aminoreban EN itself but also the bitterness-suppressing effect of the five flavors, which contained many elements such as organic acids and flavor components, and the effect of dilution (by water) on this bitterness. Using regression analysis of data derived from the taste sensor and from human gustatory data for four representative points, we were able to predict the bitterness of 50 g Aminoreban EN solutions diluted with various volumes of water (14-300 ml), with or without the addition of a selected flavor. CONCLUSIONS: Even though this prediction method does not offer perfect simulation of human taste sensations, the artificial taste sensor may be useful for predicting the bitterness intensity of elemental diets containing various flavors in the absence of results from full gustatory sensation tests.