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Biomedical subjects

N Miyoshi

Publications and source records attributed to N Miyoshi.

At least 55 records · Page 3Linked to original sources

Microfluorocytometric detection of nuclear DNA damage to cancer cells in squamous cell carcinoma after hyperthermia.

An 84-year-old woman had a squamous cell carcinoma on her left thigh. Before operation, half of the tumor mass was irradiated with a microwave at 43 degrees C for an hour. After the hyperthermia, the tumor mass was removed, and the DNA instability was investigated by microfluorocytometric measurement of the cells stained with pararosaniline-Feulgen after hydrolysis with 2N HCl at 30 degrees C. The kinetic parameters k1, which is the rate constant for the generation of an apurinic acid, k2, which is the rate constant for the depolymerization of the apurinic acid and which reflects the degree of DNA instability for acid hydrolysis, and y0, which is the theoretical value of the single-stranded DNA present initially and reflects the degree of DNA damage, were determined by a computer fitting of the Bateman function to the hydrolysis curve plotting of the fluorescence intensity of the pararosaniline-Feulgen staining the apurinic acid against the different hydrolysis times. The parameter values y0 and k2 of the cells treated by hyperthermia were much higher than those of the cells treated without hyperthermia. It is our conclusion that this microfluorometric detection would be valuable as an evaluation method for a cancer therapy, although more data from other cancers and treatments will be needed.

Aged↗

Human c-yes-1-related proto-oncogene in clinically normal dogs.

Human c-yes-1-related canine proto-oncogene in genomic DNAs from 21 clinically normal dogs was analyzed by Southern blot hybridization. The present study indicated that this proto-oncogene was well conserved in clinically normal dogs, however, there were structural changes in some dogs. These changes were different in each individuals and detected in low frequency.

Animals↗

Amplification of a c-yes-1-related oncogene in canine lymphoid leukemia.

Genomic DNAs from 4 canine spontaneous lymphoid neoplasms were examined by Southern blot hybridization using 7 kinds of oncogene probes. Analysis using a human c-yes-1 cDNA probe revealed amplification of the proto-oncogene in a DNA sample from a dog with lymphoid leukemia. Its degree was about 4- to 8-fold in comparison with a control DNA sample from a healthy dog. The genomic abnormality of canine c-yes-1-related oncogene may have a role in the tumorigenesis of this neoplasm, although the significance of the structural change in this oncogene remains unclear.

Animals↗

A case with fear of emitting body odour resulted in successful treatment with clomipramine.

A patient with fear of emitting body odour of about 20 years' duration is reported. The patient was improved with clomipramine, a tricyclic antidepressant. Generally in Japan, he would be classified as having anthropophobia. However, according to the criteria advocated by Munro and Chmara (1982), he could be classified as having monosymptomatic hypochondriasis (MH). There are some reports that antidepressants have been effective in patients with either anthropophobia or MH. Therefore, a subgroup of patients which respond to antidepressants may exist in these disorders.

Adult↗

[Clinical efficacy of arbekacin in deep MRSA infection. Including follow-up study after the termination of chemotherapy].

We analyzed the efficacy of arbekacin (ABK) using monotherapy or combined therapy on deep MRSA infection to find the most adequate usage of the drug. We also followed-up the isolation incidence of MRSA after the end of chemotherapy. The results are summarized as follows: 1. Clinical efficacy of ABK on 29 pneumonia and 3 septicemia due to MRSA was 42.9% in ABK monotherapy (9 patients), 62.5% in combined therapy with ABK and minocycline (9 patients), 100% with ABK and imipenem/cilastatin (IPM/CS) (7 patients), and 100% with ABK and other drugs (7 patients). 2. As for microbiological efficacy, combined therapy with ABK and IPM/CS or other drug was superior to other methods. Among patients from whom two or more species of bacteria were isolated, causative bacteria persisted in many cases, and some replacements occurred. 3. Kidney functions deteriorated in two patients that underwent monotherapy or combined therapy with ABK and IPM/CS, but they recovered when therapy was completed the completion. 4. In the three month follow-up study after ABK therapy, we found four cases of renewed infections after disappearance of MRSA. When just decreases in the number of MRSA resulted upon the chemotherapy, the relapse occurred in all cases. 5. Above results indicate that ABK is effective in MRSA infection, and combined therapy with beta-lactams is superior to other methods in serious MRSA infections. We also suggest that chemotherapy should be continued until the complete disappearance of MRSA is achieved.

Adult↗

Hyperthermic intravesical peplomycin perfusion treatment for bladder cancer.

Thirty-three patients with malignant bladder tumours were treated by hyperthermic intravesical perfusion. The study group included 25 patients with superficial T1 bladder tumours, 4 with T2 tumours and 4 with T3 tumours. Physiological saline solution containing 40 micrograms/ml peplomycin with/without 2% ethanol was used as a perfusate. Perfusion was carried out for 2 h 3 times per week and this regimen was repeated every other week, totalling 6 sessions in 3 weeks. Three patients achieved a complete response (CR), 5 a partial response (PR) (more than 50% tumour reduction), 8 a minimal response (MR) (25-50% tumour reduction) and the remaining 17 patients showed no response (NC). Complete, partial and minimal responses were obtained only in patients with T1 tumours. The NC patients included 4 with T2 and 4 with T3 lesions. Profuse haematuria from invasive tumour was markedly reduced after treatment. A major side effect was irritation of the bladder and urethra. Our results indicate that hyperthermic perfusion with ethanol and peplomycin is clinically safe, is accompanied by very little pain and may be useful for the management of superficial bladder tumours in elderly patients.

Aged↗

Proto-oncogene of genomic DNA, related to the human epidermal growth factor receptor (EGFR) gene, from clinically normal domestic animals.

Genomic DNAs of cattle, horses, pigs, dogs, cats and chickens were surveyed using Southern blot hybridization analysis, with a human EGFR cDNA fragment. Several bands with different numbers and molecular weights were observed under the condition of low stringency in the individual animal species. The bands showing DNA polymorphism were observed among bovine genomic PstI-digested DNAs from 4 individuals and EcoRI-digested genomic DNAs from 4 chickens. These results may provide basic data which are useful for analysis of tumorigenetic mechanisms in domestic animals.

Animals↗

Differential immunohistochemical staining of cancerous cells with anti-single-stranded DNA antiserum in ordinary pathological paraffin section after DNA-denaturation by acid hydrolysis.

Based on the finding that the nuclear DNA of cancerous cells is much more unstable than that of non-cancerous cells, yielding a larger amount of single-stranded DNA by acid hydrolysis, we developed a new method of cancer cell detection in ordinary pathological sections by immunohistochemical staining with anti-single-stranded DNA antiserum after acid hydrolysis. Methylated bovine serum albumin was conjugated with heat-denatured calf thymus DNA and used as the antigen of single-stranded DNA, and white male rabbits were immunized with the antigen to obtain the polyclonal antiserum. Ordinary paraffin-embedded sections were prepared from the formalin-fixed biopsy specimens taken from 482 malignant tumors and 73 benign tumors of human epithelial and non-epithelial origins. Additional 82 biopsy specimens of borderline malignancy were also examined. The sections were immunohistochemically stained with the antiserum after RNase digestion and DNA denaturation by hydrolysis with 2 N HCl at 30 degrees C. The acid hydrolysis for 20-30 min was optimum for the specimens fixed with 10% buffered formalin at room temperature for 16-24 hrs, and all cancerous cells were specifically stained positive, in sharp contrast to the negative stainability of all non-cancerous cells including inflammatory cells. This new method gives us decisive help in making diagnosis of malignancy in daily pathological examination. The possibility of malignancy of borderline lesion was discussed.

Animals↗

A novel class of nuclear RNA (nRNA) with a long life span as a gene repressor candidate.

Different systemic organs of fetal mice were continuously labelled with 5-3H-uridine during the organogenesis periods, and chased for various lengths of time after birth. In the autoradiographs made on paraffin-embedded sections of the organs taken after the chase for longer periods than 1 week, including a 12-months chase, specific labels were present exclusively in all the nuclei. The specific nuclear labels were resistant to RNase A or H digestion and to acid hydrolysis with 1 N HCl at 60 degrees C for 5 min, but were completely abolished by DNase digestion or prolonged acid hydrolysis for 10 min, the optimum condition for the Feulgen reaction to stain DNA. Electrophoretic analysis of the total nucleic acids extracted from the different organs chased for 3 or 12 months showed all the tritium radioactivity to be present in the DNA fraction before digestion with DNase or RNase A, and to be completely absent from the DNA fraction and shifted to the RNA fraction, or to be largely destroyed by degradation, after each digestion, respectively. By HPLC analysis of the total nucleic acid extract after further successive digestions with nuclease P1 and alkaline phosphatase into the constituent nucleotides, it was shown that all tritium activity was incorporated in uridine, without any detectable label in other nucleotides. By the simultaneous labeling of human peripheral lymphocytes at the late S-phase with 5-3H-uridine and BrdU, it was demonstrated that the autoradiographic labels, which, this time, were labile to RNase A digestion, were present in the G-bands of the spread chromosomes as identified by BrdU immunohistochemistry. The findings strongly indicate the presence of a novel class of nuclear RNA (nRNA). This type of RNA (a) may be localized in the nucleus in close association or hybridization with nuclear DNA, (b) have a life span as long as that of the cell, and (c) be duplicated in the late phase of DNA replication. The nRNA may play a fundamental role as gene repressor existing in the G-bands of metaphase chromosomes in the process of ontogeny and cytodifferentiation.

Animals↗

Chemosensitive neurons in the area postrema of the rat and their possible functions.

The present study is an attempt to demonstrate chemosensitive neurons within the area postrema (AP) electrophysiologically. Three types of chemosensitive neurons were identified: 1) glucose-responsive neurons that may participate in control of blood glucose and satiation, 2) sodium (osmotic pressure)-responsive neurons that may contribute to control of sodium and water balance of the body fluid and may be involved in salt appetite, 3) nausea-related neurons which respond to excess distension of stomach and LiCl as well. They may play a role in formation of conditioned taste aversion.

Animals↗

Integral laser photodynamic treatment of refractory multifocal bladder tumors.

Integral photodynamic therapy with hematoporphyrin derivative was performed on 35 patients who had resistant transitional cell carcinoma of the bladder, mainly carcinoma in situ. The light source was an argon ion pumped dye laser (wavelength 630 nm.) using rhodamine B. Two types of laser light scattering diffuser developed at our department were used: a motor driven laser light scattering diffuser with computer regulation, and an endoscope modified light scattering diffuser tipped with a small quartz bulb containing a lipid nutritious solution as the scattering medium. The total energy density used was 10 to 30 J./cm.2. Of the 35 patients 24 (68.6%) achieved a complete response and 5 (14.3%) a partial response at 3 months. In 10 of the 24 patients there was no recurrence with an average tumor-free interval of 20.9 +/- 16.7 months, ranging from 5 to 60 months. Bladder capacity was decreased to approximately 150 ml. for 3 months after the integral photodynamic therapy without any evidence of hydronephrosis on excretory urograms, except for 2 patients who had a contracted bladder before photodynamic therapy. Integral photodynamic therapy may prove to be useful for the treatment of carcinoma in situ of the bladder.

Adult↗

Mucosal changes of the gallbladder in anomalous union with the pancreatico-biliary duct system.

Histological examination of the gallbladder mucosa was made on a total of 39 patients with anomalous union of the pancreatico-biliary duct system. The most characteristic finding was mucosal hyperplasia of the gallbladder. Measurement of the height of the mucosa revealed that 15 cases (38.5%) showed mucosal hyperplasia composed of ordinary gallbladder epithelium without any metaplastic changes, including two cases of primary mucosal hyperplasia of the gallbladder. The other finding was metaplastic changes which were observed in 26 cases in various degrees, of whom 16 cases showed only focal metaplasia. These findings indicate that metaplasia frequently occurred in the gallbladder but the distribution of metaplasia was relatively focal in anomalous union with the pancreatico-biliary duct system. In our present materials there were nine cases of gallbladder adenocarcinoma and based on our classification, they were divided into seven cases of metaplastic type and two cases of non-metaplastic type. This ratio was not different from that of usual gallbladder carcinoma not complicated by this anomaly. These results indicate that the reflux of pancreatic juice into the gallbladder by anomalous union with the pancreatico-biliary duct system may cause two different effects on the gallbladder mucosa, the first being a proliferative effect resulting in mucosal hyperplasia and the other being chronic irritation causing metaplastic changes. The relation between these changes and the pathogenesis of gallbladder carcinoma is briefly discussed.

Adolescent↗

A new method of determining intracellular free Ca2+ concentration using Quin2-fluorescence.

We determined intracellular free Ca2+ concentration by fluorescence spectroscopy and the time-resolved measurements of 2-[(2-amino-5-methylphenoxy) methyl]-6-methoxy-8-aminoquinoline-N,N,N',N'-tetraacetic acid, tetrapotassium salt (Quin2) incorporated in suspended mouse leukemia L1210 cells. The paper reports the following two points. (1) Various fluorescence spectrum patterns in cell suspensions dissolved with Quin2 acetoxy methylester were compared with those of the complex in buffer solution containing esterase. (2) The fluorescence lifetime of Quin2 bound to Ca2+ was approx. 4.5-11 times longer (10 +/- 1 ns) than that (1.5 +/- 0.5 ns) of Quin2. The fraction of the long lifetime component was plotted against the concentration of CaCl2 in buffer solution. From the results obtained, it was found that approx. 35 nM Ca2+ was contained in each L1210 cell.

Aminoquinolines↗

Abnormal structure of the canine oncogene, related to the human c-yes-1 oncogene, in canine mammary tumor tissue.

Cellular oncogenes of genomic DNA in 6 canine primary mammary tumors were screened by Southern blot analysis, using 7 oncogene probes. A canine genomic oncogene related to the human c-yes-1 oncogene was detected as abnormal bands in solid carcinoma genomic DNA digested with EcoRI, HindIII, HindIII-EcoRI, or HindIII-BamHI. Comparison was made between other tumor specimens and control specimens obtained from 4 clinically normal dogs--1 mixed breed and 3 Shiba Inu dogs (the same breed as the dog from which the solid carcinoma was obtained). These abnormal bands were 0.1 to 1 kilobase shorter than the normal gene. However, digestion of genomic DNA obtained from normal WBC of this dog also produced all of the abnormal bands as observed in digested DNA from the solid carcinoma tissue. Therefore, in this dog, the genomic DNA of all somatic cells from the ontogenic stage still had the abnormal sequences related to the human c-yes-1 oncogene, and it is possible that this abnormal structure may have some role (eg, as an initiator) in tumorigenesis or the progression of this tumor.

Animals↗

Proto-oncogenes of genomic DNA in clinically normal animals of various species.

To provide information about oncogenes for molecular biological studies of tumors in domestic animals, the proto-oncogenes homologous to the c-myc, c-erbB-2, c-ros-1, c-yes-1, v-myc, v-Ki-ras, and v-Ha-ras oncogenes of genomic DNA in cattle, horses, pigs, dogs, cats, and chickens were investigated by Southern blot hybridization. High molecular weight genomic DNA in each of the animals contained proto-oncogenes that had a certain homology with the oncogenes used, but the extent of nucleotide homology of the proto-oncogenes differed in number and molecular weight: ie, 1 or 2 bands at 1.6 to 22.0 kilobase (kb) in the c-myc probe, 1 or 2 bands at 1.1 to 16.0 kb in the c-ros-1 probe, 1 to 3 bands at 0.7 to 23.0 kb in the c-erbB-2 probe, 1 to 4 bands at 0.6 to 18.0 kb in the c-yes-1 probe, 1 to 3 bands at 1.6 to 30.0 kb in the v-myc probe, 1 to 7 bands at 1.0 to 36.0 kb in the v-Ki-ras probe, and 1 to 4 bands at 1.0 to 27.0 kb in the v-Ha-ras probe. Furthermore, signal strength of each band, as determined by autoradiography, was not always the same for each probe in the various animals. Our findings indicate that these proto-oncogenes are well conserved with species specificities in each animal.

Animals↗

Combination effect of hyperthermia and photodynamic therapy on carcinoma.

We investigated the time course of hematoporphyrin oligomer uptake in squamous cell carcinoma transplanted into C3H mice after intraperitoneal injection by measuring its fluorescence intensity. The intensity peaked 83 hours after the injection. The tumors disappeared after an average of 4.12 days with a single application of photodynamic therapy or microwave hyperthermia 83 hours after hematoporphyrin oligomer injection. However, all the tumors treated with these single modalities regrew within 4 weeks after the treatments. Therefore, we investigated the therapeutic effect of microwave hyperthermia in combination with photodynamic therapy. As a result, the combination effect was strongest when high-dose therapy was used as the first choice, irrespective of the order of the treatments, and no tumor regrowth occurred for 45 days.

Animals↗