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Biomedical subjects

M Matsuda

Publications and source records attributed to M Matsuda.

At least 1,711 records · Page 95Linked to original sources

Assay of vitamin B6 in human plasma with graphitic carbon column.

A high performance liquid chromatographic (HPLC) procedure for measuring pyridoxal-5'-phosphate (PLP) and certain forms of B6 vitamers in plasma is presented here. This HPLC procedure consisted of a single graphitic carbon column with a fluorescence detector employing an isocratic eluent (15% acetonitrile: 1% perchloric acid: 0.05% sodium bisulfite). The graphitic carbon column is useful in acidic eluent without deteriorization. The relatively low fluorescent intensity of PLP under acidic conditions is improved by its derivatization with bisulfite in the eluent during chromatographic separation. Using this procedure, the detection limit of PLP is 50 fmol, and an aliquot of 5-50 microL of human plasma is required giving satisfactorily precise results within 5 min. We applied this method to the determination of PLP and certain B6 vitamers in human plasma after oral supplementation of pyridoxine.

Chromatography, High Pressure Liquid↗

Measurement and validity of the ventilatory threshold in patients with congenital heart disease.

The purpose of the present study was to measure oxygen uptake (VO2) at the ventilatory threshold (VT) in patients with congenital heart disease using a progressive exercise protocol on a treadmill and to evaluate the validity and feasibility of this procedure. Eight control subjects and seventeen patients performed a maximal exercise test with breath-by-breath measurement of ventilation and gas exchange variables. VT(VE) was determined by the change in the ventilatory equivalent for VO2 and carbon dioxide output, VT(V-sl) by the V-slope method, and the lactate threshold (LT) by the change in blood lactate concentration; these parameters were determined in 100%, 88%, and 96% of subjects, respectively. The interobserver error among three evaluators was not significant, and LT was correlated with each VT (r = 0.97, 0.92; p = 0.0001) and with peak VO2 (r = 0.91; p = 0.0001). The VTs were correlated with each other when expressed as milliliter per minute and milliliters per kilogram per minute. It was concluded that a progressive exercise protocol on a treadmill was a feasible procedure for determining the VTs in most individuals and that VTs were valid, useful parameters for evaluating submaximal exercise tolerance in patients with congenital heart disease.

Adolescent↗

Molecular characterisation of human campylobacteriosis in Northern Ireland: evidence of clonal stability.

BACKGROUND: No study has compared Campylobacter isolates from a human source in Northern Ireland over an extended period of time. AIM: To investigate the clonal stability of thermophilic campylobacters isolated from acute bacterial enteritis in Northern Ireland from 1992 to 1999. METHODS: Human isolates (n=272), originating from faeces, were characterised at the sub-species level using enterobacterial repetitive intergenic consensus sequence (ERIC2)--random amplification of polymorphic DNA (RAPD) typing. RESULTS: Thirteen genotypes were identified where three types, namely ERIC A, ERIC C and ERIC I, accounted for 28.3%, 14.3% and 13.6%, respectively. There were no significant associations (p>0.05) between sex, age groupings and year of isolation and ERIC2 genotype, with the exception of ERIC D, which showed a significant decline in isolation with time (p=0.019). CONCLUSIONS: ERIC genotypes were stable over this period, except genotype C which was lost during this time. Previous molecular typing methods showed campylobacters to be heterogeneous but this study suggests that the local human Campylobacter population consists of several common and stable genotypes. This study has established a database of local ERIC2 genotypes. This methodology may allow the establishment of an all-island database of clinical campylobacters that would be valuable in reducing human campylobacteriosis in Ireland.

Campylobacter↗

Developmental expression of cytochrome P450S within intrasplenically transplanted fetal hepatocytes.

Fetal hepatocytes were harvested at day 20 of gestation from spontaneously hypertensive rats (SHR) and then transplanted into recipient adult SHR spleens. Morphological examination of the recipient spleens revealed that, after 4 and 10 wk, large masses of hepatocytes were present in the red pulp with apparent cord-like structures. Larger batches of hepatocytes were observed in the spleens at 10 wk after than at 4 wk after transplantation. Of major significance was the fact that hepatocyte transplanted spleens were able to express several families of cytochrome P450 (cyto P450) proteins 2-10 wk after transplantation. Immunochemical determinations revealed that cytos P450 IA1, P450 IIB1, P450 p, P450 HLp, and P450 LA omega could be detected without any prior induction. All were intensely expressed 6 wk after transplantation; however, P450 IA1 and P450 IIB1 did not appear to be expressed by 2 wk after transplantation. Although cytos P450 p and P450 HLp did not appear to be expressed by 10 wk after transplantation, they were induced with dexamethasone at that time. Cyto P450 LA omega and peroxisomal acyl CoA oxidase were expressed 6 wk after transplantation in a 70% hepatectomized host. These results demonstrate that fetal hepatocytes can be successfully transplanted into the spleens of recipients and that the fetal hepatocytes appear to grow and develop cyto P450 metabolizing systems.

Animals↗

Gas stress test for assessment of corneal endothelial function.

PURPOSE: This study was undertaken to evaluate the endothelial pump function by monitoring both corneal swelling response under hypoxia and dehydration response following hypoxia in vivo. METHODS: Humidified nitrogen gas was used to obtain corneal swelling, and humidified gas mixed with oxygen and nitrogen was used for corneal dehydration. First, in 6 young volunteers, we investigated the most suitable oxygen level for evaluating pump function by changing oxygen levels. Then, with the optimal oxygen level, we attempted to evaluate pump function in 53 normal subjects, 5 Fuchs' dystrophy patients, and 3 iridocorneal endothelial syndrome (ICE) patients. RESULTS: Swelling rate showed similar values regardless of age, but both dehydration rate and swelling rate plus dehydration rate decreased with aging. The swelling rate of 5 guttata corneas was significantly higher than that of age-matched control corneas. In contrast, dehydration rate markedly decreased in guttata corneas, while the swelling rate plus dehydration rate of guttata corneas was comparable to that of age-matched corneas. In the 3 ICE corneas, however, swelling rate, dehydration rate, and swelling rate plus dehydration rate were markedly lower than those of both the fellow corneas and the age-matched control corneas. CONCLUSION: These observations lead us to conclude that in order to evaluate pump function, it is necessary to monitor not only dehydration response following hypoxia but also swelling response under hypoxia.

Adult↗

Tolbutamide and insulin stimulation of fructose-2,6-bisphosphate formation in hepatocytes differ.

The effects of tolbutamide and pancreatic hormones on liver fructose-2,6-bisphosphate (F-2,6-P2) formation were examined using isolated rat hepatocytes. Glucagon decreased the F-2,6-P2 level in a dose-dependent manner. Insulin (greater than 10(-9) M) increased the F-2,6-P2 level reduced by glucagon (less than 10(-9) M), but did not show a stimulatory effect on this activator formation in the absence of glucagon. On the other hand, tolbutamide increased the F-2,6-P2 level in hepatocytes regardless of the presence or absence of glucagon. Tolbutamide (2 mM) stimulation on liver F-2,6-P2 formation was enhanced by the concomitant addition of insulin (10(-8) M) in the presence of glucagon (3 X 10(-11) M). These observations suggest that the regulatory effect of tolbutamide on liver F-2,6-P2 level is independent of that of insulin.

Animals↗

Centrally induced feline emotional behavior and limbic kindling.

The threshold intensity required for electrical hypothalamic/mesencephalic stimulation to elicit emotional behavior in cats was examined chronologically during limbic kindling (n = 9/5), and without kindling (n = 3/3) for a matching control period. After the completion of primary site amygdaloid (AM) or hippocampal (HIPP) kindling, the hypothalamic defensive behavior threshold showed a +/- 20% fluctuation from the prekindling baseline with a mean tendency for reduction as in the control animals. The flight behavior threshold increased in the kindled animals, and decreased in the controls; however, the degree of change was not statistically significant. In contrast, the mesencephalic defensive behavior threshold, which was quite stable in the controls, decreased significantly upon primary (mean -13.5%) and secondary site (mean -22.2%) AM kindling, whereas the flight and screeching (sound emission) behavior threshold remained unchanged throughout kindling. The decreased or unchanged thresholds for each behavior were independent of the timing of the hypothalamic/mesencephalic stimulation after kindled generalized convulsion. The decreased mesencephalic defensive behavior threshold persisted during a 1- to 3-month rest period without kindling stimulation. These findings suggest that acquisition of feline AM-kindled seizure susceptibility is associated with a lasting facilitation of midbrain (but not hypothalamic) excitability for defensive (but not flight) behavior.

Animals↗

In vivo antibacterial activity of a prodrug of NM394, a thiazetoquinoline carboxylic acid derivative.

NM394 (6-fluoro-1-methyl-4-oxo-7-(1-piperazinyl)-4H-[1,3]-thiazeto [3,2-a]quinoline-3-carboxylic acid) has potent, broad-spectrum antibacterial activity in vitro, but not in vivo. To increase the bioavailability of NM394, various prodrugs were synthesized and tested. One of them, NM441, an N-(5-methyl-2-oxo-1,3-dioxol-4-yl) derivative, showed potent in vivo antibacterial activity. Using thin-layer chromatography-bioautography, we confirmed that after oral administration, NM441 was readily absorbed and hydrolyzed to NM394. Other prodrugs of NM394 were only partially metabolized to NM394. In pharmacokinetic studies in mice and monkeys, we found that the blood levels of NM394 were 7.8 and 5.9 times greater, respectively, when NM441 rather than NM394 was administered. These findings suggest that NM441 is an effective prodrug of NM394.

Animals↗

Feasibility of measuring 5-fluorouracil catabolic potential by oral loading.

The efficacy of 5-fluorouracil (5-FU) treatment and the incidence of adverse events differ among patients and depend to some extent on individual variations in drug catabolism. This feasibility study aimed to determine the optimum conditions for a 5-FU oral load test, which would allow the simple evaluation of individual differences in 5-FU catabolism. Patients with colon cancer were given oral 5-FU (200 mg/day) for 3 days (n = 36) or a single 100 mg dose (n = 14). Serum concentrations of uracil, dihydrouracil, 5-FU and 5-fluoro-5,6-dihydrouracil were measured before and after 5-FU administration. The results suggested that a decline in 5-FU metabolism was associated with continuous administration and increasing age. We conclude that a continuous load of 5-FU is necessary in order to predict the efficacy and side-effects of the drug. The 3-day regimen, with its ease of administration, merits further study to assess its possible clinical application.

Administration, Oral↗

Greater development of 1,2-dimethylhydrazine-induced colon cancer in a rat model of type 2 diabetes mellitus.

Several clinical cohort and case-control studies have suggested a link between diabetes and colon cancer. Otsuka Long-Evans Tokushima Fat (OLETF) rats spontaneously develop type 2 diabetes mellitus and Long-Evans Tokushima Otsuka (LETO) rats are non-diabetic. The relationship between type 2 diabetes mellitus and colon cancer was examined in these rats. The carcinogen 1,2-dimethylhydrazine was administered subcutaneously once weekly for 10 weeks, and the animals were killed and necropsied in week 29. All OLETF rats and 80% of the LETO rats developed cancer. The number of colon cancers per rat was significantly greater in the diabetic than in the non-diabetic rats. Although the tumours tended to be larger in diabetic rats, the difference was not statistically significant. No significant differences were observed in the depth of invasion or histological type of cancer in the two groups. Type 2 diabetes mellitus may enhance the generation and growth of colon cancer.

1,2-Dimethylhydrazine↗

Long term (five-year) survival following radical surgical treatment plus adjuvant chemotherapy (FAM) in advanced gastric cancer: a controlled study.

Several drugs and their associations are being used for adjuvant or complementary chemotherapy with the aim of improving results of gastric cancer treatment. The objective of this study was to verify the impact of these drugs on nutrition and on survival rate after radical treatment of 53 patients with gastric cancer in stage III of the TNM classification. A control group including 28 patients who had only undergone radical resection was compared to a group of 25 patients who underwent the same operative technique followed by adjuvant polychemotherapy with FAM (5-fluorouracil, Adriamycin, and mitomycin C). In this latter group, chemotherapy toxicity in relation to hepatic, renal, cardiologic, neurological, hematologic, gastrointestinal, and dermatological functions was also studied. There was no significant difference on admission between both groups in relation to gender, race, macroscopic tumoral type of tumor according to the Borrmann classification, location of the tumor in the stomach, length of the gastric resection, or response to cutaneous tests on delayed sensitivity. Chemotherapy was started on average, 2.3 months following surgical treatment. Clinical and laboratory follow-up of all patients continued for 5 years. The following conclusions were reached: 1) The nutritional status and incidence of gastrointestinal manifestation were similar in both groups; 2) There was no occurrence of cardiac, renal, neurological, or hepatic toxicity or death due to the chemotherapeutic method per se; 3) Dermatological alterations and hematological toxicity occurred exclusively in patients who underwent polychemotherapy; 4) There was no significant difference between the rate and site of tumoral recurrence, the disease-free interval, or the survival rate of both study groups; 5) Therefore, we concluded, after a 5-year follow-up, chemotherapy with the FAM regimen did not increase the survival rate.

Antineoplastic Combined Chemotherapy Protocols↗

[Smoking prevalence at the Federal University of São Paulo, 1996--preliminary data of an institutional program].

PURPOSE: To evaluate the prevalence of tobacco smoking among health workers, professores, nurses and students of Federal University of São Paulo and the acceptance of an institutional program for quitting smoking. METHODS: We analized the answers of a questionnairy with 51 questions, distributed to people from different categories. RESULTS: The total percentage of answered questionnaires was 48.6% (2613). The answers obtained from health workers were 37.3%, professors 49.0%, nurses 52.7% and students 76.5% The total percentage of smokers at UNIFESP was 15.5%:23.7% for health workers, 18% for professors, 16% for nurses and 8.6% for students. There was no significant statistical difference between the prevalence of smoking among females (17.3%) and males (16.3%). The age of major prevalence of smoking was between 31 to 40 years (26.6%). For all ages, people who have university level smoked less, independent of sex. Eighty three percent (83%) of the UNIFESP workers and students are worried of being passive smokers. Between the smokers, 55% were thinking of quiting and 42% declared that need some help to quit. CONCLUSION: We concluded that educational programs and help for cessation at institutional level are necessary and well accepted at UNIFESP, and the completion of these programs will contribute to the obeying of the prohibitive laws of no smoking within the community.

Adolescent↗

Follow-up study of the heart in acromegaly: pre- and post-operative evaluation.

Pre-operative and post-operative echocardiographic data were analyzed from 8 patients with acromegaly. Pre-operatively, end-diastolic diameter was greater than 55 mm in 5 patients (63%) and concentric left ventricular hypertrophy was observed in 3 patients (38%). However, left ventricular function was normal (fractional shortening of the left ventricle greater than 28%) in all patients except 1. All patients had increased left ventricular mass. There was no significant correlation between left ventricular mass and basal plasma growth hormone concentration. An average of 23.9 months after hypophysectomy, growth hormone concentration was significantly decreased. However, the abnormal echocardiographic findings remained. In conclusion, echocardiographic abnormalities (left ventricular dilatation and hypertrophy) are common in patients with acromegaly, but systolic function is, in general, maintained. These cardiac abnormalities persist after reduction of plasma growth hormone concentration.

Acromegaly↗

In situ detection of prolactin receptor mRNA and apoptotic cell death in mouse uterine tissues with adenomyosis.

Prolactin receptor (PRLR) mRNA was visualized by in situ hybridization in adenomyotic uteri of mice with ectopic pituitary grafting. The signals of PRLR mRNA were mainly detected in the smooth muscle cells of myometrium and the epithelial cells of uterine glands and lumen. In the uteri with adenomyosis, the expression intensity was stronger than that in the normal uteri of control mice. This might reflect the higher serum prolactin (PRL) levels in the mouse with pituitary graft, as reported previously. Electron microscopic observation showed that apoptotic cell death occurred in certain cells in the myometrium bearing adenomyotic changes. Such changes were rarely observed in the normal uteri. The relevance of these findings to the development of adenomyosis was examined.

Animals↗

Identification of insertion mutations in HIV-1 reverse transcriptase causing multiple drug resistance to nucleoside analogue reverse transcriptase inhibitors.

OBJECTIVE: A novel 2-amino acid insertion between codons 69 and 70 of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) which confers multiple drug resistance has recently been reported. Independently, we have identified similar insertion mutations in Japanese hemophiliacs and attempted to analyze their emergence in conjunction with therapy regimens and their contribution to drug resistance using recombinant technology. METHODS: The plasma and peripheral blood mononuclear cells (PBMCs) of 348 HIV-1-infected hemophiliacs were screened for HIV-1 RT mutations relevant to nucleoside analogue inhibitors and isolating viruses. Contribution of each insertion to drug resistance was studied by introducing the mutations into a T-cell line-tropic NL4-3 infectious clone and testing the drug susceptibilities of the recovered virus. RESULTS: Insertion of the 2-amino acid residue was found in 4 of the 348 cases and was strongly associated with prolonged chemotherapy with zidovudine (AZT) and didanosine (ddI). The virus isolated from 1 of the 4 cases possessed the same insertion. Characterization of these virus and the recombinant NL4-3 with the insertion strongly suggested that the insertion caused resistance not only to AZT and ddI but also to lamivudine (3TC) and zalcitabine (ddC). CONCLUSION: A 2-amino acid insertion between codons 69 and 70 of RT was detected in 4 of 348 (1.1%) Japanese hemophiliacs and was found to be associated with multiple drug resistance to nucleoside analogue RT inhibitors.

Amino Acid Sequence↗