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Biomedical subjects

N Ishida

Publications and source records attributed to N Ishida.

At least 271 records · Page 15Linked to original sources

Evaluation of recombinant human tumor necrosis factor by scheduled intratumoral administration in mice bearing transplantable tumors.

The antitumor effect of recombinant human tumor necrosis factor (rTNF) was examined against Meth A fibrosarcoma in BALB/c mice and Sarcoma-180 in ddY mice. Significant hemorrhagic necrosis in tumor tissues occurred within 24 hr when optimal rTNF (1,000 to 5,000 units per mouse) was injected intratumorally on day 5 after intradermal inoculation of 5 x 10(5) tumor cells. Complete tumor regression resulted when two repeated courses of administration a week, each for 3 consecutive days, were given. For this marked effect to occur, however, initial tumor weight should not be greater than 1 g. When the initial tumor was greater than 1 g the surgical removal of tumor tissues was conducted and followed by rTNF administration. This caused hemorrhagic necrosis and the regression was the case with smaller tumors. When the cured mice were rechallenged with same tumors, more than 60% of mice rejected the tumors in a specific manner. In spite of such demonstration of specific immunity, well-known immunological effector mechanisms such as augmentation of natural killer cell activity, activation of antibody dependent cellular cytotoxicity or induction of interferon activity by rTNF were not detected in normal and tumor-bearing mice, suggesting that the activation of immunoregulatory cells by TNF itself may not involve at least in an early stage of TNF treatment. These results suggest that rTNF is a potent therapeutic agent for a certain solid tumor when the protocol of administration is optimized.

Animals↗

Suppressive effect of 1-methyladenosine on the generation of chemiluminescence by mouse peritoneal macrophages stimulated with opsonized zymosan.

The preparation of an in vitro assay system for the immunosuppressive activities of modified nucleosides on macrophage (M phi) functions is described. Briefly, Listeria-elicited mouse peritoneal M phi s (Lm-M phi s) were treated with nucleosides in vitro for 18 hr at less than 1 mM concentration and chemiluminescence (CL) was measured after stimulation with opsonized zymosan. To confirm the usefulness of this assay (in vitro test), the immunosuppressive activities of 1-methyladenosine (m1 Ado) and adenosine (Ado) were determined by mouse Listeria infection (in vivo test), CL generation by M phi s obtained from the peritoneal cavity of nucleoside-treated mice (in vivo-in vitro test), and the in vitro test. The immunosuppressive activity of m1 Ado detected by the in vitro test was confirmed by the in vivo and the in vivo-in vitro tests. As for Ado, no immunosuppression was detected by the in vivo and the in vivo-in vitro tests through a potent suppressive effect was detected by the in vitro test. The ineffectiveness of Ado can be explained by the in vivo conversion of Ado to an inactive form. Thus, the proposed in vitro test seems to be useful, with the provision that the known in vivo metabolism is taken into consideration.

Adenosine↗

Effects of glycyrrhizin (SNMC: stronger Neo-Minophagen C) in hemophilia patients with HIV infection.

Glycyrrhizin (GL) not only has an inhibitory effect on HIV replication but also exhibits interferon-inducing and natural killer (NK)-enhancing effects and improves liver dysfunction. Thus, large doses of GL (200-800 mg/day) were intravenously administered for more than 8 weeks to 9 hemophilia A patients with HIV infection (asymptomatic carrier, AC). Lymphocyte count increased in all 9 cases. OKT4 OKT8 ratio was elevated in 6 out of the 9 cases and OKT4-positive lymphocytes increased in 8 out of the 9 cases; 66.7% and 88.9% improvement, respectively. Changes in NK cell activity and mitogenic responsiveness to PHA, Con A and PWM were not significant. Liver dysfunction, noted in 4 cases, clearly improved. Serum electrolytes, protein, lipids, and renal function were within normal levels and no serious side-effects were observed during treatment. On the other hand, in 3 cases of hemophilia without HIV infection, the number of OKT4 lymphocytes was not significantly altered during treatment. From these results, large dose administration of GL to HIV-positive hemophilia patients (AC) seems to be effective in preventing development of AC into AIDS by raising the number of decreased OKT4 lymphocytes and improving liver dysfunction.

Acquired Immunodeficiency Syndrome↗

Sensitive detection of two IgG Fc receptors of mouse macrophages by chemiluminescence analysis.

Luminol-enhanced chemiluminescence assay was used to detect the surface expression and the consequent activation of receptors (FcRI and FcRII) of murine macrophages (M phi s). When murine IgG2a was used for the specific detection of FcRI and IgG2b for FcRII, a newly established procedure enabled us to detect the activation of each receptor with as few as 3 X 10(5) M phi s. Briefly, TNP-SRBC coated with monoclonal IgG2a or IgG2b antibodies directed to TNP (sensitized SRBC) were used as reagent, in the presence of 1 X 10(-5) M luminol, and the emission was measured with a liquid scintillation counter. When results obtained by chemiluminescence counting were compared to the results obtained by the rosette formation by adding the same SRBC reagent to peritoneal M phi s obtained after ip injection of Listeria, fortified chemiluminescence counting allowed us to obtain a more definite answer about the activation of each receptor. Under the conditions established, the specific activation of FcRI was obtained by the addition of rIFN alpha A/D to the resident M phi s in vitro and the specific activation of spleen M phi FcRII by iv injection of IAP (Immunosuppressive acidic protein) into mice. These two results supported the independence of the two receptors detected by the assay.

Animals↗

[The evaluation of simple estimation method of prostate size by transabdominal ultrasound].

We evaluated 107 patients who were diagnosed as benign prostatic hypertrophy preoperatively and underwent transurethral resection of the prostate. We obtained the maximum cross section of the adenoma by transabdominal scanning using a real-time large-sized convex scanner and a 3.5 MNz. transducer. Regarding the adenoma as an ellipsoid with revolution around its transverse axis, 2 diameters (breadth and height) were measured with transverse scan (a and b, respectively). We then calculated the values of the estimated size of the adenoma according to the formula for a spheroid. V = (pi/6) ab2. Thus the calculated weight were correlated with the resected weight of the specimen after a complete transurethral resection of the prostate. As a result, the sonographic estimation provided a high correlation coefficient: r = 0.956. The result suggested that our method of transabdominal ultrasonography in the preoperative evaluation of the prostatic size was rapid, simple and noninvasive, was well adapted for screening sufferers from prostatic diseases, and gave preoperative information for transurethral resection of the prostate and other surgical operation.

Aged↗

Antiviral activity of carboxyethylgermanium sesquioxide (Ge-132) in mice infected with influenza virus.

The protective effect of carboxyethylgermanium sesquioxide (Ge-132) in mice infected with a mouse-adapted strain of influenza virus (H2N2) was investigated. When mice were exposed to a 10 LD50 dose of influenza virus via aerosol and were treated orally with 20 or 100 mg/kg of Ge-132 daily for 6 consecutive days, a significant protective effect was demonstrated. The antiviral effect of Ge-132 was indicated by an increase of survivors, a prolongation of mean survival days, an inhibition of the development of lung consolidation, and a decrease of virus titer in lung tissues, as compared to infected control mice treated with phosphate-buffered saline. Natural killer (NK) cell activity in the spleens and lungs of the infected mice was also significantly augmented after the oral administration of Ge-132. In addition, NK cells stimulated with Ge-132 in vivo showed killing activity against NK-insensitive Meth-A cells infected with influenza virus. Because no virucidal or virustatic activities of Ge-132 on the virus were found in vitro, this protective effect in mice against influenza virus infection may be displayed through immunomodulating activities of this compound such as the augmentation of NK cell activity.

Animals↗

[The quantitative determination of cyanide by FTD-GC].

In recent buildings, new materials containing chemical goods, chemical fibers and the like are in common use. Some of them evolve hydrogen cyanide (HCN) when burning. When a person meets a fire, therefore, it is necessary to measure his blood concentration of HCN as well as that of CO-Hb%. The blood concentration of HCN can be measured by colorimetry such as pyridine-pyrazolone method, gas-chromatography, electrode method, or the like. However, these methods require much time in pretreatment and the preparation of the reagent. We have investigated to find a new measurement method free from the above demerits, so that we have found the method by FTD-GC (Flame Thermionic Detecter Gas-chromatograph) is convenient and can be effected in a short time in comparison with the aforementioned method. GC-7AG made by Shimadzu Co. is employed as gas-chromatography and the headspace method is used in the quantitative determination. Samples are employed as solutions of potassium cyanide (KCN) either in distilled water or in fresh blood. The concentration of KCN is gradient in the range of 0.5-6 micrograms/ml. The quantitative conditions is investigated by a solution in distilled water as standard. Using this method, the concentration of HCN in the blood sample is determined slightly lower than that in the distilled water; however, this method is advantageous in convenience and the saving of time. Therefore, the method is useful in practice for the determination of the concentration of HCN.(ABSTRACT TRUNCATED AT 250 WORDS)

Chromatography, Gas↗

[Spectrophotometric determination of boric acid by the curcumin method].

We have contrived an improved curcumin method for a spectrophotometric determination of the boric acid content, suitable for use on biologic materials to determine cases of poisoning. The use of this method enables detection of boric acid from a level of 10 micrograms/ml up to 5 mg/ml. The steps of this measurement method follow. Initially, boric acid was extracted by using a modification of Agazzi's method, i.e., to 1 ml of the sample solution, 0.2 ml of a 50% solution of sulfuric acid is added, along with 4 ml of 10 v/v% 2-ethyl-1,3-hexanediol/chloroform (an EHD solution). This mixture was then shaken for 5 min and then centrifuged for 10 min at 3,000 rpm. The extract of this chloroform phase was dehydrated with anhydrous sodium sulfate and used for the coloring reaction sample. The colorimetry procedure for determining the boric acid content follows. Fifty microliters of the extract solution was placed into a dry tube, to which 0.5 ml of a 0.3% curcumin/acetic acid solution and 50 microliters concentrated sulfuric acid were added, and the contents mixed thoroughly. The reaction mixture was then allowed to stand for 30 min at room temperature. (Rosocyanin is formed by the reaction of boric acid and protonated curcumin). Next, ethanol (3-138 ml) was added to the reaction mixture to decompose the excess protonated curcumin. Then, the absorbancy of the resulting solution was measured at 550 nm against a blank test solution. Ethanol was added to enable the measurement of the absorbancy (ethanol amounts tested were 3, 6, 12, 24, and 138 ml).(ABSTRACT TRUNCATED AT 250 WORDS)

Boric Acids↗

[Determination of boric acid in biological materials by curcuma paper].

The field of legal medicine has seen a recent increase of poisoning by boric acid and, in cases of emergency, a simple method of making a qualitative analysis of the boric acid content is a necessity. Thus, we have examined curcuma paper (turmeric paper) to see if it can provide a qualitative analysis of the boric acid content in biological materials, so as to identify cases of poisoning. It was found that curcuma paper can provide a preliminary analysis of the quantitative content of boric acid, and that about 0.1 mg/ml of boric acid can be determined. The steps for this testing method follow. First, either blood or urine is acidified with a 6 N concentration of hydrochloric acid, i.e., in the case of urine, 0.5 ml of urine is added to 0.1 ml hydrochloric acid, and for blood, 0.5 ml of blood is added to 0.2 ml hydrochloric acid. If the sample is not sufficiently acidic, more hydrochloric acid is added. Next, a drop of the sample is placed on the curcuma paper and, after drying at room temperature, a red stain results if boric acid is present. (Rosocyanin is formed by the reaction of boric acid and protonated curcumin). Then, a 1 N concentration of sodium hydroxide is dropped onto the stained place, and if rosocyanin is present, the stain will turn blue. Informatively, to make curcuma paper, filter paper (No. 2) is soaked in a saturated curcumin/ethanol solution and then air dried.(ABSTRACT TRUNCATED AT 250 WORDS)

Boric Acids↗

Corneal endothelium in a case of mitochondrial encephalomyopathy (Kearns-Sayre syndrome).

A case of mitochondrial encephalomyopathy (Kearns-Sayre syndrome) with corneal endothelial abnormality is reported. A 22-year-old woman had retinitis pigmentosa, external ophthalmoplegia, complete heart block, ataxia, muscle weakness, dementia, sensorineural hearing loss, and was of short stature. Renal dysfunction, diabetes mellitus, and amenorrhea were also observed. Biopsy revealed decreased cytochrome c oxidase (complex IV) activity in muscle mitochondria. The corneal endothelium examined by specular microscope showed decreased cell density, severe polymegathism, and pleomorphism in both eyes. To our knowledge, this is the first report concerning primary corneal endothelial abnormality in a case with mitochondrial encephalomyopathy. The corneal endothelium is one of the tissues that could be affected by the enzyme deficiency present in this disease.

Adolescent↗

[The influence of endotracheal tube size on the airway flow resistance and work in infants analyzed using a simulation technique].

The respiratory flow velocity of 74 infants under halothane-nitrous oxide-oxygen anesthesia was measured, and the statistical relationship between the age in months and the flow velocity was obtained with least square method. The flow resistance of endotracheal tubes (size of 2.5, 3.0, 3.5, 4.0, 4.5 and 5.0 mmI.D.) with the flow velocity below 45 ml.sec-1 was also measured in in vitro experiments. The standard respiratory flow velocity of 6, 12, 24, 36, 48 and 60 month old infants was estimated by the recurrent formula. The mean flow resistance and respiratory work with each size of endotracheal tube were calculated by simulation technique using these data. These data coincided well with the data obtained by calculation based on in vitro experiments.

Aging↗

Four species of cDNAs for cytochrome P450 isozymes immunorelated to P450C-M/F encode for members of P450IID subfamily, increasing the number of members within the subfamily.

A chicken antibody mono-specific to cytochrome P450C-M/F, which exists in untreated male and female rat liver and catalyses the 2- and 16 alpha-hydroxylation of estrogens (1), was used to screen a cDNA library of male Sprague-Dawley rat liver. Four cDNA clones which encoded P450 isozymes, CMF1a, CMF1b, CMF2 and CMF3, were isolated. CMF1a and CMF2 deduced consisted of 504 and 500 amino acid residues, respectively, while C-terminal 487 and 324 residues for CMF1b and CMF3, respectively, were deduced from the 5'-truncated cDNAs. The isozymes were more than 72% similar in amino acid sequences to each other and to rat P450db1, P450db2 (2), and to a mouse male specific C-P45016 alpha (3), suggesting that they belonged to a new P450 subfamily, P450IID. CMF1a and db1, and CMF2 and db2, respectively, were 99.2% and 99.0% similar in amino acid sequences, suggesting that they were virtually identical. CMF1a and CMF1b were different but 96.1% similar, and CMF3 was between 76% and 78% similar to other members of the rat P450IID family.

Amino Acid Sequence↗

A single amino acid substitution converts cytochrome P450(14DM) to an inactive form, cytochrome P450SG1: complete primary structures deduced from cloned DNAS.

Genes for lanosterol 14-demethylase, cytochrome P450(14DM), and a mutated inactive cytochrome P450SG1 were cloned from S. cerevisiae strains D587 and SG1, respectively. A single nucleotide change resulting in substitution of Asp for Gly-310 of cytochrome P450(14DM) was found to have occurred in cytochrome P450SG1. In this protein the 6th ligand to heme iron is a histidine residue instead of a water molecule, which may be the ligand for the active cytochrome P450(14DM). Molecular models of the active sites of the cytochrome P450(14DM) and cytochrome P450SG1 were built by computer modeling on the basis of the known structure of that of cytochrome P450CAM whose crystallographic data are available. The mechanisms which may cause a histidine residue to gain access to the heme iron are discussed.

Amino Acid Sequence↗

Adoptive immunotherapy by pantropic killer cells recovered from OK-432-injected tumor sites in mice.

A murine malignant ascites model with BAMC-1 tumors was established previously, which was cured completely by five consecutive i.p. injections of OK-432. We have found that peritoneal mononuclear cells from these animals contained antitumor effector cells which could destroy nonspecifically a variety of tumor cells in vitro. They were tentatively called pantropic killer cells (PKCs). The present study was essentially designed to show the antitumor effectiveness of the PKCs in vivo by the use of an adoptive immunotherapy model. The growth of BAMC-1 tumors transplanted s.c. 5 days earlier was significantly suppressed by passive transfer of 5 x 10(6) to 2 x 10(7) PKCs induced by injection of OK-432 into BAMC-1 bearing donor mice, while more than 1 x 10(8) immune spleen cells from the same donors treated with OK-432 were required to achieve the similar effects. Furthermore, if the tumor-bearing recipients were pretreated with 180 mg/kg of cyclophosphamide 1 h before the adoptive transfer, even 5 x 10(6) PKCs could induce complete regression of the tumors transplanted 5 days earlier. This protocol made it possible even to achieve the complete regression of larger tumors (9-10 mm in diameter) in recipients transplanted 12 days earlier. The PKCs were, as expected, able to cure not only BAMC-1-bearing animals but also Meth-A-bearing mice. As effector cells for adoptive immunotherapy, therefore, the PKCs induced by OK-432 seem to be as effective as, if not better than, lymphokine-activated killer cells expanded in vitro by culturing tumor infiltrating lymphocytes with interleukin-2. Although the study on surface markers of PKCs did not unequivocally discriminate these from lymphokine-activated killer cells, the present findings are considered significant indicating that a potent biological response modifier such as OK-432 can induce pantropic killer cells which are extremely effective in destroying various tumor cells in vivo. One of the advantages of OK-432 therapy over lymphokine-activated killer cell therapy, therefore, is that the former does not require the tedious and time-consuming in vitro procedures which are essential for the latter.

Animals↗

Cloning and sequence analysis of cDNA encoding the precursor of a human endothelium-derived vasoconstrictor peptide, endothelin: identity of human and porcine endothelin.

A cDNA encoding a human endothelium-derived vasoconstrictor peptide, endothelin, was isolated from a human placenta cDNA library. The nucleotide sequence of this cDNA clone showed that the primary structure of the human preproendothelin has 212 amino acid residues and is highly homologous to porcine preproendothelin, and that human endothelin is identical with porcine endothelin.

Amino Acid Sequence↗