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Biomedical subjects

S Fujimoto

Publications and source records attributed to S Fujimoto.

At least 343 records · Page 19Linked to original sources

Macrophage migration inhibitory factor in the human ovary: presence in the follicular fluids and production by granulosa cells.

Cytokines play an important role in ovarian function. We unexpectedly found high expression of macrophage migration inhibitory factor (MIF) mRNA in human ovarian tissues. Hence, we examined the presence of MIF in the follicular fluid because the follicular microenvironment is important for oocyte fecundity. The follicular fluids were collected from ovaries of patients undergoing in vitro fertilization and embryo transfer. A higher amount of MIF was identified in the follicular fluid, 80.3 +/- 4.6 ng/ml (mean +/- SE), in which the concentration was significantly decreased as the size of the follicles increased. To detect MIF mRNA expression in the granulosa cells, reverse transcription-polymerase chain reaction was carried out and this showed an amplified transcript specific for MIF. Furthermore, the presence of MIF protein in the granulosa cells was confirmed by Western blot analysis. These results suggest the possibility that MIF mediates various immunological events in the process of oocyte development.

Cells, Cultured↗

Restriction fragment length polymorphism analysis and random amplified polymorphic DNA analysis of Campylobacter jejuni strains isolated from patients with Guillain-Barré syndrome.

Campylobacter jejuni serotype O19 strains associated with the Guillain-Barré syndrome (GBS) and other strains were examined by restriction fragment length polymorphism (RFLP) analysis of polymerase chain reaction products of the flaA genes and by random amplified polymorphic DNA (RAPD) analysis. RFLP analysis showed that regardless of LIO serotype, geographic origins, or association with GBS, the O19 isolates shared an identical digestion pattern by each of four restriction endonucleases, DdeI, MboI, MseI, and AluI. In contrast, among C. jejuni O1 or O2 strains, RFLP patterns were different even among strains of the same LIO serotype. The results of the RAPD analysis were consistent with the flaA RFLP data. These data indicate that all of the O19 strains that were tested were closely related to one another whether they were or were not associated with GBS.

Animals↗

Atrial conduction abnormalities in patients with systemic progressive sclerosis.

BACKGROUND: Atrial abnormalities in patients with progressive systemic sclerosis have not been evaluated in terms of intra-atrial conduction. We hypothesized that a delay in atrial conduction in these patients might produce diastolic abnormalities as well as atrial arrhythmias. OBJECTIVE: To evaluate the atrial function of patients with progressive systemic sclerosis by using echocardiography to measure the intra-atrial electromechanical activation coupling interval. METHODS: Twenty patients with progressive systemic sclerosis were assessed by Doppler echocardiography. Twenty age-matched healthy controls were also evaluated. Two-dimensional guided M-modes of ventricular long axes were recorded using simultaneous phono- and electrocardiograms of the apical four chamber view at the right lateral, septal and left lateral sites of the atrioventricular rings. Transmitral and tricuspid pulsed Doppler flow velocities were also recorded. Filtered P wave duration was measured on the signal averaged ECG to determine the duration of atrial electrical activation. RESULTS: There was a delay in P on the electrocardiogram (P) at the onset of atrial contraction on long axis M-modes at all three atrioventricular ring sites in patients with progressive systemic sclerosis as compared with controls (P-right; 56 +/- 13 vs 47 +/- 10 ms, P-septal; 74 +/- 14 vs 55 +/- 10 ms, and P-lateral; 93 +/- 16 vs 72 +/- 11 ms, P < 0.01). Inter-atrial conduction time [(P-lateral)-(P-right)] was delayed in patients with progressive systemic sclerosis, compared with healthy controls (37 +/- 15 vs 25 +/- 6 ms, P < 0.01). Mitral A waves acceleration and deceleration times were also decreased in the patients. The interval was prolonged between P to the onset and the peak of the A wave in transmitral flow. Duration of the filtered P wave was significantly prolonged in progressive systemic sclerosis as compared with controls (124 +/- 12 ms vs 106 +/- 8 ms, P < 0.01). PQ intervals, E waves and acceleration and deceleration times did not differ significantly in progressive systemic sclerosis vs, controls. The A wave acceleration rate on transmitral flow (peak A wave velocity/acceleration time) showed a significant correlation with inter-atrial conduction delay (r = 0.55, P < 0.01). CONCLUSIONS: Intra-atrial electromechanical coupling intervals were delayed in patients with progressive systemic sclerosis. Thus, the mechanical late diastolic filling time due to atrial contraction in the total diastolic phase was severely limited, and this resulted in a restricted mitral A wave. We should therefore evaluate patients with progressive systemic sclerosis for significant atrial abnormalities.

Atrial Function↗

Prediction of drug cytotoxicity in 9L rat brain tumor by using flow cytometry with a deoxyribonucleic acid-binding dye.

OBJECTIVE: Flow cytometry (FCM) with a deoxyribonucleic acid (DNA)-binding dye, propidium iodide, provides a rapid and quantitative method to detect apoptotic cell death. This technique was used to examine the sensitivity of tumor cells to anticancer agents, as a novel test of chemosensitivity in vitro. METHODS: The in vitro chemosensitivity of 9L gliosarcoma cells to a panel of anticancer agents (cisplatin, nimustine, adriamycin, cyclophosphamide, vincristine, 5-fluorouracil, and methotrexate) was investigated by both FCM, yielding DNA histograms, and a microtiter tetrazolium test, measuring cellular metabolism. Clinically achievable concentrations of the agents were used for the analysis of DNA histograms and proliferation of 9L cells in vitro. Rats intracranially inoculated with 9L cells were treated with the agents, and tumor masses were visually monitored by using magnetic resonance imaging with gadolinium-diethylenetriaminepentaacetic acid enhancement. RESULTS: The cytotoxic effect of anticancer agents examined by the microtiter tetrazolium test correlated with a decreased G0/G1 peak in the DNA histograms. Serial FCM analysis showed that the decrease in the G0/G1 peak was subsequently accompanied by increased hypodiploid areas, suggesting DNA fragmentation induced by the agents. The in vitro chemosensitivity test and cell proliferation examination showed that all agents except cisplatin were effective. Growth retardation of inoculated brain tumors and prolonged survival of inoculated rats were observed with treatment with the anticancer agents, except cisplatin. CONCLUSION: The present study shows that FCM analysis with a DNA-binding dye can detect DNA damage induced by anticancer agents, and it suggests that this technique is a novel method to test chemosensitivity in vitro.

Animals↗

Induction of specific cytotoxic T lymphocytes against autologous brain tumor by crossreactive allo-tumor cell stimulation.

Cytotoxic T lymphocytes (CTL) against autologous malignant brain tumor were generated in peripheral blood lymphoid cells (PBL) prepared from a patient with a malignant brain tumor by stimulation of the cultured PBL for 7 days with attenuated crossreactive malignant melanoma (MM2) cells pretreated with mitomycin C. The crossreactive MM2 cells were effective for antigen stimulation for CTL induction in place of autologous glioblastoma cells, which are difficult to expand in culture. The optimal ratio between nylon wool-passed T lymphocytes and nylon wool-adherent accessory cells to induce CTL in the patient's PBL was found to be 25 to 1. In vitro-activated CTLs induced by MM2 were cytotoxic not only to MM2, but also to the autologous tumor cells in an HLA class I-restricted manner, and their surface phenotype was found to be CD3+ and CD8+. CTL therapy using cross-reactive allogeneic tumor cells as the stimulator could be clinically valuable to treat malignant brain tumors.

Aged↗

Effect of exercise training and detraining on gas exchange kinetics in patients with chronic obstructive pulmonary disease.

Symptom-limited incremental exercise tests are used to estimate the training effect on patients with chronic obstructive pulmonary disease (COPD). However, there is a need for objective parameters for measurement on submaximal exercise testing. The purpose of this study was to assess the usefulness of measurement of oxygen uptake (VO2) kinetics during a constant work rate exercise test of patients with COPD after exercise training. Eleven patients with COPD performed exercise tests before and after cycle ergometer training on 3 days per week for 8 weeks; they then went without training for 5 months and performed the same tests. They performed an incremental exercise test to symptom-limited maximum and a constant work rate exercise test for 6 min on a cycle ergometer. The time constant of VO2 during the onset of constant work rate exercise was significantly decreased (from 63.5 +/- 7.8 s to 53.2 +/- 8.0 s) after exercise training (P = 0.021), but was significantly increased (to 73.4 +/- 14.9 s) after 5 months without training (P = 0.001). The oxygen pulse at steady state during constant work rate exercise testing was significantly increased after exercise training but decreased 5 months later. The change in blood lactate from rest to steady state during constant work rate exercise was significantly decreased after exercise training, but increased 5 months later. Measurement of the time constant of VO2 and oxygen pulse during constant work rate exercise are useful for the objective evaluation of the training effect of patients with COPD.

Aged↗

Chromosomal analysis in 894 induced abortuses from women of advanced maternal age in relation to gestational weeks and fetal sex ratio.

OBJECTIVE: To clarify the prenatal sex ratio, and types and frequencies of chromosome anomalies in relation to gestational weeks in women of advanced maternal age. METHODS: Chromosomal analysis was performed in 894 induced abortuses derived from elderly pregnant women 35 or more years of age. RESULTS: The embryonic sex ratio in the chromosomally normal (abnormal) gave a preponderance of females over males in lower gestational weeks [366/458 (63/73); 79.9 to 100 (86.3 to 100) in 4-10 weeks]; the reversal of dominancy in those from females to males occurred in the 11th gestational week [41/29(5/4); 141.4 to 100 (125.0 to 100) in 11-20 weeks]. The frequency of trisomy in females at 10 weeks of gestation was significantly high (7/28; 25%). CONCLUSIONS: This study showed that the existence of a difference of and a reversal of the sex ratio in gestational weeks. It was considered that there might be a lower viability of genetically handicapped males as compared to such females. However, the reversal of the sex ratio, such as that observed between 9th and 10th gestational weeks, might reflect events occurring after implantation.

Abortion, Induced↗

Screening for bacterial vaginosis and cervicitis aimed at preventing premature delivery.

OBJECTIVE: We performed screening and treatment of bacterial vaginosis and cervicitis during pregnancy for prevention of premature delivery. The incidence and the etiology of lower genital tract infections and premature delivery were examined to elucidate the effectiveness and problems of this trial. METHODS: Three hundred sixty-five pregnant women who attended the obstetrical outpatient clinic in our hospital were included as subjects. The screening was done at prenatal-care visits around the 10th, 20th, and 30th gestational weeks. RESULTS: Fifty-eight women were diagnosed as bacterial vaginosis and 54 women were successfully treated. Thirty-five women were diagnosed as cervicitis and all were cured. Nine women (2.5%) delivered before the 37th gestational week and 1 women delivered before the 35th gestational week because of cervical incompetence. CONCLUSIONS: The treatment of bacterial vaginosis and cervicitis may prevent microbes in the lower genital tract from penetrating the intrauterine cavity and thereby reduce the incidence of premature delivery.

Adolescent↗

Importance of the transitional zone between the cervical stroma and the parametrium in the treatment of cervical carcinoma.

OBJECTIVES: Parametrial invasion is an important risk factor in cervical carcinoma. Because the vessels of the parametrium start in the zone between the cervical compact stromal layer and the parametrium, which we have defined as the parametrial initial zone (PIZ) or transitional zone, the invasion of a carcinoma into this part of the cervix is also important. Our objectives in this study were find a characteristic index of the transitional zone and to clarify the relationship between a transitional-zone invasion and the prognosis in a case. METHODS: Thirty-three cases of carcinoma in situ were available to use for the morphological examination of the cervix. The cervix was divided into 8-12 sections. In each section, the outer diameters of the short axis of the arteries were measured with a light microscope and a micrometer. Using the arterial diameter as the characteristic index of the transitional zone, we performed epidemiological studies on 312 patients with clinical Stage Ib-III squamous-cell carcinoma of uterine cervix. RESULTS: The arteries with an axis exceeding 300 microns were concentrated in the transitional zone at a rate of 94.6%. Therefore, arteries of this size are used as the characteristic index of the transitional zone. Patients with invasion to transitional zone only had lymph node metastasis at the rate of 28.1%, compared with 0% for patients who had no permeation to the zone (p < 0.001). The 5-year survival rate of patients with invasion to the transitional zone only was 87.3%, which is significantly lower than those without invasion (99.2%) (p < 0.01). CONCLUSIONS: Measuring the short axis (300 microns < or = ) of the artery is the best method by which to pinpoint the transitional zone. The invasion of a carcinoma into the transitional zone in the cervix should be treated as a parametrial involvement.

Adult↗

The behavior of endometrial hyperplasia: a prospective study. Endometrial Hyperplasia Study Group.

OBJECTIVE: To clarify the behavior of endometrial hyperplasia in a prospective study. METHOD: Fifty-one patients with endometrial hyperplasia were followed up for 6 months. Samples of endometrial tissues were taken by uterine endometrial biopsy every 4 weeks during the first 3 months and at the end of follow-up. RESULTS: In 69% (35/51) of the patients histological picture of the endometrium became normal during the observation period. The lesions persisted in 17% (6/35) of the patients with simple hyperplasia, in 25% (1/4) of those with complex hyperplasia, in 14% (1/7) of those with simple atypical hyperplasia, and in 80% (4/5) of the patients with complex atypical hyperplasia. In the remaining 3 patients with simple hyperplasia, the lesions progressed to complex atypical hyperplasia by the end of follow-up, after showing a normal endometrium. CONCLUSION: Most cases of endometrial hyperplasia, except for complex atypical hyperplasia, disappeared spontaneously within a short period of time.

Adult↗

Comparison of an improved AFP kit with the intra-amniotic PSP dye-injection method in equivocal cases of preterm premature rupture of the fetal membranes.

OBJECTIVE: We developed an improved AFP kit for detecting AFP in leaked amniotic fluid. We studied the relationship between the results of the improved AFP kit and those of the intra-amniotic PSP dye-injection test (PSP test), and compared the results of two tests with the prognosis for subsequent pregnancy outcome, in order to know the diagnostic value of the two tests for preterm PROM, especially in equivocal cases. METHODS: We compared the results of the AFP kit test with those of the PSP test in 8 equivocal cases of preterm PROM (18th to 34th weeks of gestation). RESULTS: The results of the AFP kit test coincided with those of the PSP test in all 8 equivocal cases of preterm PROM. The 4 cases that tested positive resulted in preterm deliveries; and the other 4 cases that tested negative resulted in full-term deliveries. CONCLUSION: The use of the AFP kit test is useful in assessing the prognosis for subsequent pregnancy in equivocal cases of preterm PROM. The rapid and accurate AFP kit test for diagnosing preterm PROM can be substituted for the PSP test in bedside and outpatient-clinic assessments.

Adult↗

High-level plasmid-mediated gentamicin resistance and pheromone response of plasmids present in clinical isolates of Enterococcus faecalis.

Eleven pheromone-responding plasmids encoding erythromycin or gentamicin resistance were isolated from multiresistant clinical Enterococcus faecalis isolates. The plasmids were classified into six types with respect to their pheromone responses. The three erythromycin resistance plasmids responded to different pheromones. Of the eight gentamicin resistance plasmids, four plasmids responded to same pheromone. Southern hybridization studies showed that the genes involved in regulation of the pheromone response were conserved in the drug resistance plasmids.

Anti-Bacterial Agents↗

Cloning and genetic analysis of the UV resistance determinant (uvr) encoded on the Enterococcus faecalis pheromone-responsive conjugative plasmid pAD1.

The conjugative pheromone-responsive plasmid pAD1 (59.6 kb) of Enterococcus faecalis encodes a UV resistance determinant (uvr) in addition to the hemolysin-bacteriocin determinant. pAD1 enhances the UV resistance of wild-type E. faecalis FA2-2 and E. faecalis UV202, which is a UV-sensitive derivative of E. faecalis JH2-2. A 2.972-kb fragment cloned from between 27.7 and 30.6 kb of the pAD1 map conferred UV resistance function on UV202. Sequence analysis showed that the cloned fragment contained three open reading frames designated uvrA, uvrB, and uvrC. The uvrA gene is located on the pAD1 map between 28.1 and 29.4 kb. uvrB is located between 30.1 and 30.3 kb, and uvrC is located between 30.4 and 30.6 kb on the pAD1 map. The uvrA, uvrB, and uvrC genes encode sequences of 442, 60, and 74 amino acids, respectively. The deduced amino acid sequence of the uvrA-encoded protein showed 20% homology of the identical residues with the E. coli UmuC protein. Tn917 insertion mutagenesis and deletion mutant analysis of the cloned fragment showed that uvrA conferred UV resistance. A palindromic sequence, 5'-GAACNGTTC-3', which is identical to the consensus sequence found within the putative promoter region of the Bacillus subtilis DNA damage-inducible genes, was located within the promoter region of uvrA. Two uvrA transcripts of different lengths (i.e., 1.54 and 2.14 kb) which terminate at different points downstream of uvrA were detected in UV202 carrying the deletion mutant containing uvrA. The longer transcript, 2.14 kb, was not detected in UV202 carrying the deletion mutant containing both uvrA and uvrB, which suggests that uvrB encodes a terminator for the uvrA transcript. The uvrA transcript was not detected in any significant quantity in UV202 carrying the cloned fragment containing uvrA, uvrB, and uvrC; on the other hand, the 1.54-kb uvrA transcript was detected in the strain exposed to mitomycin C, which suggests that the UvrC protein functions as a regulator of uvrA.

Adenosine Triphosphatases↗

Cloning and genetic and sequence analyses of the bacteriocin 21 determinant encoded on the Enterococcus faecalis pheromone-responsive conjugative plasmid pPD1.

The pheromone-responsive conjugative plasmid pPD1 (59 kb) of Enterococcus faecalis encodes the bacteriocin 21 (bac21) determinant. Cloning, transposon insertion mutagenesis and sequence analysis of the bac21 determinant showed that an 8.5-kb fragment lying between kb 27.1 and 35.6 of the pPD1 map is required for complete expression of the bacteriocin. The 8.5-kb fragment contained nine open reading frames (ORFs), bacA to bac1, which were oriented in the same (upstream-to-downstream) direction. Transposon insertions into the bacA to bacE ORFs, which are located in the proximal half of bac21, resulted in defective bacteriocin expression. Insertions into the bacF to bac1 ORFs, which are located in the distal half of bac21, resulted in reduced bacteriocin expression. Deletion mutant analysis of the cloned 8.5-kb fragment revealed that the deletion of segments between kb 31.6 and 35.6 of the pPD1 map, which contained the distal region of the determinant encoding bacF to bac1, resulted in reduced bacteriocin expression. The smallest fragment (4.5 kb) retaining some degree of bacteriocin expression contained the bacA to bacE sequences located in the proximal half of the determinant. The cloned fragment encoding the 4.5-kb proximal region and a Tn916 insertion mutant into pPD1 bacB trans-complemented intracellularly to give complete expression of the bacteriocin. bacA encoded a 105-residue sequence with a molecular mass of 11.1 kDa. The deduced BacA protein showed 100% homology to the broad-spectrum antibiotic peptide AS-48, which is encoded on the E. faecalis conjugative plasmid pMB2 (58 kb). bacH encoded a 195-residue sequence with a molecular mass of 21.9 kDa. The deduced amino acid sequence showed significant homology to the C-terminal region of HlyB (31.1% identical residues), a protein located in the Escherichia coli alpha-hemolysin operon that is a representative bacterial ATP-binding cassette export protein.

Amino Acid Sequence↗

Lectin affinity electrophoresis in a yolk sac tumour in the vagina with yolk sac tumour-type glycoform of alpha fetoprotein.

AIMS: To investigate a potential diagnostic use of alpha fetoprotein (alpha FP) isoform analysis by lectin affinity electrophoresis to distinguish between endodermal sinus tumours arising in the vagina in infants from those at other sites. METHODS: alpha FP in the serum of a patient with a vaginal endodermal sinus tumour was analysed for its isoforms by lectin affinity electrophoresis. The isoforms were compared with that of cord serum, sera of hepatoid adenocarcinoma of the uterus, and endodermal sinus tumour of the ovary. RESULTS: The isoforms of alpha FP obtained by lectin affinity electrophoresis in the serum of the patient with vaginal endodermal sinus tumour differed from the isoforms of alpha FP in the cord serum of normal neonates, and sera of patients with hepatoid adenocarcinoma of the uterus or endodermal sinus tumour of the ovary. CONCLUSIONS: Endodermal sinus tumour arising in the vagina could be distinguished from that in the ovary by the lectin affinity electrophoresis, and a potential diagnostic use of alpha FP isoform analysis by the lectin affinity electrophoresis for the detection of the endodermal sinus tumour in infants was demonstrated.

Diagnosis, Differential↗

Effects of hyperosmotic NaCl and glycerol stress on stress response of human HeLa cells.

The maintenance of intracellular osmotic pressure is of fundamental importance for cell survival. Since osmoregulatory processes are important to all living organisms, large fluctuations in environmental osmolarity may elicit or modulate stress responses. We examined whether hyperosmotic stress induced or modulated stress responses in human HeLa cells. When HeLa cells were incubated in medium supplemented with 50-150 mM NaCl or 0.2-1.0 M glycerol for 3-27 h, the stress response, analyzed at the levels of HSP70 synthesis, HSP70 mRNA accumulation, and heat shock transcription factor (HSF) activation, was not induced by either hyperosmotic stress. In hyperosmotic 150 mM NaCl or 1.0 M glycerol medium, the stress response to heat shock was inhibited at the levels of HSF activation, HSP70 mRNA accumulation, and HSP70 synthesis. In vitro activation of HSF showed that inhibition of this activation by hyperosmotic NaCl or glycerol stress was not irreversible. Furthermore, addition of the physiological osmolyte betaine to medium reversed the inhibition of heat-induced HSP70 synthesis under hyperosmotic NaCl stress but not under hyperosmotic glycerol stress. The effect of betaine against hyperosmotic NaCl stress was observed mainly at the translation level, and betaine seemed to enable cells to translate HSP70 mRNA specifically under heat shock conditions by restoring protein synthetic ability.

Betaine↗

A case of dilated cardiomyopathy with abnormal atrioventricular conduction 5 years before the appearance of left ventricular systolic dysfunction.

The first manifestation of dilated cardiomyopathy is usually dyspnea as a result of left heart failure. In this report we describe an unusual patient with third-degree atrioventricular (AV) block unaccompanied by left ventricular dysfunction. Severe left ventricular dysfunction occurred 5 years after the implantation of a permanent pacemaker. We conclude that AV block should not be overlooked as an initial clinical manifestation of dilated cardiomyopathy.

Adult↗

Heart rate variability and ventricular arrhythmia in clinically stable patients with hypertrophic cardiomyopathy.

To investigate whether autonomic nervous dysfunction exists and to determine the relationship between autonomic nervous activity and ventricular arrhythmias in clinically stable patients with hypertrophic cardiomyopathy (HCM), we studied heart rate variability (HRV) in 21 patients with HCM and 10 age-matched healthy control subjects. Patients were divided into 2 groups: HCM I (14 patients without nonsustained ventricular tachycardia, defined as 3 or more consecutive ventricular premature beats) and HCM II (7 patients with nonsustained ventricular tachycardia). HRV was measured on 24-h ECG monitorings using both nonspectral and spectral methods (fast Fourier transform, FFT). We assessed autonomic nervous activity based on high-frequency (HF; 0.15-0.40 Hz, an index of parasympathetic nervous activity) and the low- to high-frequency power ratio (L/H ratio; an index of sympathetic nervous activity) during daytime and night-time separately. There were no significant differences in any index of HRV between the HCM I and control groups. HF in the HCM II group was significantly lower than that of both the HCM I and control groups during the day and at night, whereas the L/H ratio at night-time was significantly higher in the HCM II group than in the HCM I and control groups. These data demonstrate that, in clinically stable HCM patients without nonsustained ventricular tachycardia, autonomic nervous activity was not different to that of healthy control subjects, whereas in HCM patients with nonsustained ventricular tachycardia, parasympathetic nervous activity was reduced throughout the 24-h period, and sympathetic nervous activity at night was increased compared with healthy control subjects. Thus, HCM patients with nonsustained ventricular tachycardia have to be treated even if they do not exhibit significant clinical manifestation.

Adult↗