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Biomedical subjects

N Yoshida

Publications and source records attributed to N Yoshida.

At least 451 records · Page 25Linked to original sources

Purification and complex formation analysis of a cysteine proteinase inhibitor (cystatin) from seeds of Wisteria floribunda.

Seeds of Wisteria floribunda contain several kinds of cysteine proteinase inhibitor (cystatin). We purified and characterized one of these inhibitors, named WCPI-3. The molecular weight of WCPI-3 was estimated to be 17,500 and 15,700 by gel filtration and SDS-PAGE, respectively. The isoelectric point was 5.7. WCPI-3 formed an equimolar complex with native papain and the dissociation constant was estimated to be 6.1 nM. Complex formation between WCPI-3 and Cys25-modified papain, such as S-carboxy-methylated or S-carbamoylmethylated papain, could not be observed by gel filtration or native PAGE analysis. A peptide fragment derived from WCPI-3 digested by Achromobacter proteinase (lysyl endopeptidase) had the amino acid sequence of VVAGVNYRFVLK. The VVAG sequence in this fragment corresponds to the conserved sequence QVVAG which is considered to be one of binding regions to cysteine proteinases. The amino acid sequence of the amino-terminal portion (34 residues) of WCPI-3 was highly homologous to that of oryzacystatin from rice seeds.

Amino Acid Sequence↗

Purification and characterization of an acidic amino acid-specific endopeptidase of Bacillus subtilis obtained from a commercial preparation (Protease Type XVI, Sigma).

An acidic amino acid-specific endopeptidase was purified from Protease Type XVI (Sigma), a commercial product from culture filtrate of Bacillus subtilis, by a series of column chromatographies on CM-Toyopearl (Fractogel) and Mono-S, guided by activity assay using Boc-Ala-Ala-Pro-Glu-pNA as a substrate. The final preparation was homogeneous on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and reversed-phase HPLC. The molecular weight of the protease was estimated to be 18,000 by gel filtration on TSK gel G3000SWXL column using 6 M guanidine hydrochloride as an eluent, and 17,000 by SDS-PAGE in the presence of 2-mercaptoethanol. The isoelectric point of the protease was 7.7. Studies on the substrate specificity with peptide p-nitroanilides and natural peptides revealed that the protease hydrolyzes the peptide bonds on the carboxyl-terminal side of acidic amino acids, especially of glutamic acid. The protease was completely inactivated by DFP, indicating the serine protease nature of the protease. The activity of the protease was also inhibited by EDTA and GEDTA, and reactivated by Ca2+. The protease contained 1.3 +/- 0.2 mol/mol protein of Ca2+. These results suggest that Ca2+ plays a vital role in the protease activity.

Amino Acid Sequence↗

Role of oxygen-derived free radicals in the pathogenesis of gastric mucosal lesions in rats.

The role of oxygen-derived free radicals and lipid peroxidation in the pathogenesis of acute gastric mucosal erosion was investigated in rat models produced by burn shock stress, by treatment with regional hyperthermia, platelet activating factor, and compound 48/80, and by ischemia-reperfusion. In all experimental models, the increase in the gastric erosions and in TBA reactants in the gastric mucosa were significantly inhibited by the treatment with superoxide dismutase (SOD) and/or catalase. Pretreatment with allopurinol, a competitive inhibitor of xanthine oxidase, prevented considerably the gastric injury (a) induced by burn shock, (b) produced by treatment with compound 48/80, and (c) caused by ischemia-reperfusion. By the treatment with anti-rat neutrophil antibody, the gastric mucosal injuries induced by regional hyperthermia, platelet activating factor, and compound 48/80 were significantly inhibited; however, burn shock and ischemia-reperfusion injuries were not inhibited. These results suggest that oxygen-free radical and lipid peroxidation contribute to the formation of gastric mucosal lesions, and that the sources of oxygen radicals seem to be different among these experimental models.

Allopurinol↗

An application of magnet and magnetic sensor: measurement system for tooth movement.

A measurement system for tooth displacement and rotation in the sagittal and frontal planes was designed and tested. Four small magnetic sensors were arranged at each corner of a rectangle on a plastic sheet. They were then placed in the mouth and fixed to the front teeth with a plastic splint. A powerful magnet made of a rare earth metal was fixed to the target tooth and placed at the center of the sensors. Movement of the magnet was detected by the four sensors as the tooth trajectory. This system was tested by measuring first molar movement in human subjects with a load generated by an orthodontic aid. This system was small enough to fit in a human oral cavity and did not interfere with the orthodontic aid. Although the oral environment with high humidity and high temperature was not agreeable for the sensors, resolution was estimated at better than 3 microns including the effects of system drift. System calibration was done extra orally and distortion was calculated at less than 6% in a square of 200 microns. The accuracy of this system was not affected by temperature due to the application in the oral cavity at a relatively constant temperature. The advantage of this system was not only the two-dimensional measurement but also rotational measurement in which a 0.05 degree resolution was calculated.

Calibration↗

A facile method for preparation of t-butyloxycarbonylamino acid p-nitroanilides.

A series of p-nitroanilides of t-butyloxycarbonylamino acids, including Boc-Trp, Boc-Asn, Boc-Gln, Boc-Ser(But), Boc-Thr(But), Boc-Asp(OBut), Boc-Lys(TFA), and Boc-His(Boc), were prepared conveniently by the mixed anhydride method using 2,2-dimethylpropanoic chloride (pivaloyl chloride). The products were obtained in 40-60% yields after purification by column chromatography on Sephadex LH-20. p-Nitroanilide of histidine was purified after deprotection of Boc-His(Boc)-pNA and obtained as H-His-pNA.2HCl.

Amino Acids↗

[A case of fatal sepsis due to Vibrio vulnificus].

A 66-year-old male with chronic alcoholic liver injury was admitted on July 27, 1986 to our hospital with complaints of high fever, convulsion and skin erythema. He had consumed raw fish 3 days before, and had a scratch wound over the right arm and left leg because he had slipped in a small stream in the woods the day before admission. He was already in shock state with sepsis of V. vulnificus and DIC on admission. Although the treatment with ABPC, CP, CAZ, MINO for sepsis, and Heparin & Antithrombin III for DIC was immediately begun, he died only 10 hours after admission. On autopsy, the skin lesion revealed phlegmon with necrotizing angitis and the liver showed fatty changes with Mallory's body. The causative organism was detected from the blood and on autopsy from the skin wound, bile juice, liver, spleen, kidney and bone marrow, and its type was determined as a V. vulnificus serovar 4. It was suspected that the route of infection in this case was the raw fish rather than via the wound because the water in which he had been wounded was fresh water and the bacterium was not detected from the water, shells, nor moss existing there.

Aged↗

Physical training in patients with valvular heart disease after surgery.

The necessity, the methodology and clinical benefit of physical training were evaluated in 85 patients with valvular heart surgery. In 21 patients who had undergone the exercise stress test before surgery, corrective valvular surgery increased maximal oxygen uptake (max.VO2) from 15.4 before to 18.9 ml/min/kg, while 11 (52%) of them failed to increase exercise capacity over 5 METs. Of the total 85 patients, 41 (58%) again showed reduced exercise capacity in the initial postoperative stress test, suggesting that a return to work might be difficult without rehabilitation in these patients. We then compared two training programs: program A of short duration and frequent exercise and program B of longer duration and less frequent exercise, both using a bicycle exercise set at an intensity of 70% of the max.VO2. Both programs similarly increased max.VO2, while patients preferred program A, suggesting that exercise of longer duration could not be tolerated because of deconditioning. Program A was then prescribed to 62 patients, and it increased max.VO2 from 18.2 to 20.7 ml/min/kg after 4 weeks training without any complication. In 9 patients who served as controls undergoing no physical training, no spontaneous improvement in exercise capacity was observed. Of the 76 patients who received either program A or B, 28 patients failed to increase the max.VO2 by 10% or more. These patients presented atrial fibrillation, a cardiothoracic ratio greater than or equal to 60% or exercise-induced ST depression more frequently, suggesting that residual cardiac dysfunction might inhibit the training effects.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Production and some properties of monoclonal antibodies against human pancreatic secretory trypsin inhibitor.

Hybridomas that secrete monoclonal antibodies against human pancreatic secretory trypsin inhibitor (PSTI) were established by fusion of spleen cells obtained from mice immunized with PSTI with mouse NS-I-Ag 4/1 myeloma cells. One of three resulting monoclonal antibodies (KN-1) was found to recognize the N-terminal moiety of the inhibitor, while the others (KN-2 and KN-3) reacted with other as yet undefined parts of the molecule. Trypsin inhibitory activity of PSTI treated with KN-1 monoclonal antibody was the same as that of PSTI itself, thus indicating no relationship between the N-terminal moiety of the PSTI molecule and its inhibitory activity. We further examined the applicability of one of the monoclonal antibodies (KN-1) for immunohistochemical study of human pancreatic cancer tissue including the normal as a model, and found granular staining of the cytoplasm of the normal acinar and duct cells and also of that of adenocarcinoma cells in formalin-fixed, paraffin-embedded tissue sections.

Adenocarcinoma↗

[A case of hypogammaglobulinemia associated with polyarteritis nodosa presenting a variety of symptoms in childhood].

A case of polyarteritis nodosa (PN) in childhood involving various organs such as the gastrointestinal tract, skin, CNS, kidneys and liver with hypogammaglobulinemia is reported. This 6 month old girl was admitted to our hospital with vomiting, diarrhea, bloody stools with mucous and weight loss. For the past 5 months she had these abdominal symptoms. She was diagnosed as having PN of the Kussmaul-Maier variety on the grounds of the biopsy of skin lesion where a necrotizing vasculitis was found. Prednisolone and methylprednisolone pulse treatment were not effective in suppressing the progress of the disease. At the age of 1 year 7 month a combination therapy of prednisolone and immunosuppressants (cyclophosphamide) was started and this was found to be effective. She was discharged when she was 2 year and 2 month. The dosage of prednisolone was tapered as the activity of the PN decreased and she did well with a maintainance dosage of 9.5 mg/day. At 3 year 6 month of age she suddenly developed hypertension (the plasma renin activity was found to be 16.6 ng/m/hr. and the aldosterone 220 ng/dl). CNS involvement such as spinal cord dysfunction, left sided convulsions, cerebral hemorrhage developed 5 months later. Methylprednisolone pulse therapy was performed 3 times and 2 mg/kg/day of prednisolone was administered. In spite of this therapy she passed away with a massive cerebral hemorrhage at the age of 4 year 8 month. Unfortunately an autopsy was not performed. Results of the immunological tests proved that the hypogammaglobulinemia was a common variable immunodeficiency (CVI). It has been reported that primary immuno-deficiency syndrome is often associated with collagen disease and auto-immune disease. This lack of the defense mechanism against the virus or extra antigen could be related to the onset of collagen and auto-immune disease. As the correlation between CVI and PN has not been clarified this case is of interest as concerns the cause of PN.

Agammaglobulinemia↗

[Follow-up study on anti-HBs levels in vaccines after two and three doses of HB vaccine (1)].

UNLABELLED: An HB prophylaxis vaccination that included a primary and secondary vaccination was carried out on persons working at this university hospital. In the primary vaccination, the subjects were inoculated the second time with a vaccine derived from human blood plasma obtained from the Kitazato Therapeutic Research Institute. The third time, they were inoculated with a vaccine from the Chemo-Sero Therapeutic Research Institute derived from a second-generation vaccine organized ferment. The vaccine used for the second inoculation was obtained from the Chemo-Sero Therapeutic Research Institute. RESULTS: A total of 254 subjects were inoculated with the HB prophylaxis vaccination. 1. Of the 216 subjects inoculated with the primary vaccination, at the time of the second inoculation, 65 subjects (30%) tested positive for antibodies 3-4 months following inoculation. There was an especially high rate among females between 20 and 50 years. Thirty-two subjects (15%) tested positive 15-16 months following inoculation with a high rate among females. 2. Among 132 subjects inoculated the third time with the primary vaccination (those who tested negative in the second inoculation of the above vaccine), 76 subjects (58%) tested positive for antibodies 8-9 months following inoculation with a generally high rate among females. Thirty-five subjects (27%) tested positive 15-16 months after inoculation with a high rate among females between 20 and 30 years and among males between 40 and 60 years. 3. Among the 38 subjects who were inoculated the second time with the secondary vaccination, 8 subjects (21%) tested positive for antibodies 5 months following inoculation, with a high rate among females in their 20's.

Adult↗

[Experimental study of the trajectories of the upper first molar with extraoral force. Practical measurement of the movements in the three planes by a magnet-magnetic sensing system].

The upper first molar trajectory under the loading of extraoral force was studied in human subjects. The five loading directions, that is horizontal, upper, lower, high-pull and cervical were tested. Using a magnet-magnetic sensing system, the rectilinear and rotational movements were traced in the three planes, i.e. sagittal, frontal, and horizontal planes. It was concluded; 1. The molar inclined buccally with the loading in all the applied directions. 2. New evidence was found in the horizontal plane, i.e. the molar rotated mesially with the loading in the lower direction. The rotation may depend on the direction of the loop at the terminal of the inner-bow. Therefore it was suggested that the form of the loop affected the molar movements. 3. With the loading in the horizontal direction, the distal movement of the molar was evident and small displacement was observed in the extruded direction. 4. Loading both in upper and lower directions, the rotation of the molar was more evident than the rectilinear displacement. It was suggested that these loading would not be applicable to control the tooth in the rectilinear movements. 5. With the high-pull loading, the molar inclined mesially and shifted distally a little. On the other hand, it was greatly intruded. The results suggested that effective distal and intrusive movements would be expected by bending the outer-bow downward, since this procedure would prevent the tooth from tipping mesially. 6. The loading in the cervical direction caused large distal movement of the molar. Extrusion was also evident as it was expected from the loading direction.

Dental Stress Analysis↗

[Follow-up study on anti-HBs levels in vaccinees after two and three doses of HB vaccine. (2)].

An HB prophylaxis vaccination that included a primary and secondary vaccination was carried out on persons working at the university hospital. In the primary vaccination group, the subjects were inoculated the second time with a vaccine derived from human blood plasma obtained from the Kitazato Therapeutic Research Institute. The third time, they were inoculated with a vaccine from the Chemo-Sero Therapeutic Research Institute derived from a second-generation vaccine organized ferment. The vaccine used for the second inoculation group was obtained from the Chemo-Sero Therapeutic Research Institute. Results were summarized as follows: 1) The primary vaccination group: (1) The number of the 153 subjects inoculated with the primary vaccination at the time of the second inoculation, 49 subjects (32%) tested positive for antibodies 3-4 months following inoculation. Thirty-two subjects (20%) tested positive 15-16 months following inoculation with a high rate among females. (2) Among 104 subjects inoculated the third time with the primary vaccination (those who tested negative the second inoculation of the above vaccine), 61 subjects (58.7%) tested positive for antibodies 8-9 months following inoculation with a generally high rate among females. Twenty-eight subjects (26%) tested 15-16 months after inoculation with a high rate among females between 20 and 30 years and among males between 40 and years. 2) The secondary group: Among the 38 subjects who were inoculated the second time with the secondary vaccination, 8 subjects (21%) tested positive for antibodies 5-6 months following inoculation, with a high rate among females in their 20's. 3) The rate of antibodies formation.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

[Basic investigation of saliva SIgA].

Saliva, which contributes to maintaining health in the oral environment, is affected by the physiological functions of the organism and their condition. Dental caries and periodontosis, both of which are oral diseases with frequent occurrence, require the presence of bacteria to occur. SIgA (secretory IgA), an immune globulin that is found in large quantities in substances secreted during salivation, has an antibacterial action. Thus, in conducting a basic investigation on the relationship between these factors, the author investigated what effect the speed of saliva secretion, a physiological factor of the saliva component, has on the SIgA value. Results showed that, regarding the relationship between the salivary SIgA value and the speed of salivary secretion, a negative correlation exists although this is weak in terms of relevance. The fact that the correlation displayed a tendency toward scattering indicates a need to pay attention when measuring the elements in the saliva to those factors that evidently have an effect on the composition of the saliva.

Humans↗

[An erythremia with acquired HbH disease and chromosomal abnormality].

A 56-year-old male was admitted to the Nihon University Hospital because of general fatigue and anemia on September 21st, 1985. He had mild hepato-splenomegaly. Hematological findings showed RBC 286 x 10(4)/microliters, Hb 6.0/dl, reticulocyte count 2.5%, platelet count 9.3 x 10(4)/microliters and WBC 2,400/microliters. An erythroblast per 100 leukocytes counted in a blood film was found. Bone marrow was erythroid hyperplasia with megaloblasts. The erythroblasts were PAS positive but not ringed sideroblasts. Other laboratory data including hemolysis were all negative. This case seemed to be diagnosed as refractory anemia (RA) according to the FAB classification. Chromosomal analysis of marrow cells, however, all revealed 46, XY, 20q- at diagnosis and 46, XY, 7q- 20q- after 22 months. Furthermore, Hb electrophoresis ahd family study indicated the presence of acquired HbH disease. Neither erythroid bursts (BFU-e) nor late erythroid progenitors (CFU-e) were detected. He has had progressive anemia without proliferation of blasts for over 2 years. From these findings, we postulate that the entity of erythremia should be distinguished from RA including many heterogeneous diseases.

Chromosome Deletion↗

[The use of Pugh score to evaluate the prognostic and therapeutic influence of drinking in patients with decompensated liver cirrhosis].

The aim of this study is to evaluate the clinical usefulness of Pugh score in the prognosis of the patients (pts.) with decompensated liver cirrhosis (DLC). To determine the influence of heavy drinking with respect to the prognosis of DLC patients, 128 cases of DLC were divided into two groups including the 31 heavy drinker group (Group A) and 97 non-alcoholic group (Group B). At the time of admission, the both groups showed similar Pugh score. However, after initial two hospitalized weeks, Group A started to show significantly lower points of Pugh score than that of in Group B. Fifteen pts. (48%) in Group A and 55 pts. (57%) in Group B had greater than 9 in Pugh score at the time of admission. In Group A, 10 of 15 pts. declined the scores after initial two weeks and 9 of them lived more than one year after admission. In contrast, none of those 55 pts. in Group B declined their Pugh score after admission and 45 of them died within one year. It was concluded that Pugh score was extremely useful to evaluate the prognosis of DLC pts., and DLC pts. who had greater than 9 Pugh score were seemed to be critical. It was suggested that DLC pts. due to heavy drinking tend to improve Pugh score comparing to no alcoholic DLC pts.

Adult↗

Guidelines for histopathologic diagnosis of plantar malignant melanoma. Two-dimensional coordination of maximum diameters of lesions and degrees of intraepidermal proliferation of melanocytes.

Detection of early lesions of malignant melanoma is highly important for the improvement of the prognosis. However, diagnosis of early malignant melanoma is not easy and is often discordant even among expert dermatopathologists. In this study, we propose guidelines for the histopathologic diagnosis of plantar malignant melanoma and malignant melanoma in situ on the sole. They are formulated with two-dimensional coordination of maximum diameters of the lesions and degrees of intraepidermal proliferation of solitary-arranged melanocytes. The guidelines are simple and reproducible, and surely contribute to the accurate diagnosis of early plantar malignant melanoma.

Cell Division↗

[Clinical study of male patients with primary biliary cirrhosis (PBC)].

We studied 10 male and 23 female patients with PBC to determine whether the clinical and histological features of this disease differed in male and female patients. There were no significant difference between men and women in age distribution and biochemical examinations. In female patients, autoimmune associated conditions such as sicca syndrome, Raynaud syndrome and arthritis were observed 22%, 13% and 36%, respectively. By contrast, no male patients developed those conditions. 80% of the male patients and 70% of the female patients belonged to asymptomatic PBC, and early histological stage, such as Scheuer's I and II were observed 90% of the male patients and 78% of the female patients, respectively. No male patients showed clinical or histological progression during follow-up period (median was 64 months). Nevertheless, not a few female patients showed progression including 3 cases who died during the follow-up period (median was 47 months). We concluded that male patients with PBC tend to have favorable prognosis comparing to female patients.

Adult↗