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Biomedical subjects

M Fukunaga

Publications and source records attributed to M Fukunaga.

At least 19 recordsLinked to original sources

A flow cytometric DNA analysis of giant cell tumors of bone including two cases with malignant transformation.

BACKGROUND AND METHODS: Flow cytometric DNA analysis was performed on 30 cases of giant cell tumor (GCT) of bone with the use of paraffin-embedded sections. RESULTS: According to the criteria of Huvos, they were classified histologically into three groups: Grade 1, 26 cases; Grade 2, 4 cases; and Grade 3, no cases. Among the Grade 1 cases, 21 were diploid and 5 were aneuploid. Of the four Grade 2 cases, three were diploid and one was aneuploid. Nine patients had local relapses. Among four patients with complications by lung metastases, two have remained well at 18 and 157 months with the metastases. The other two patients, who had Grade 1 DNA diploid GCT of the 11th thoracic spine, had malignant transformation (osteosarcoma) resulting from radiation therapy. In one patient, the primary lesion exposed to radiation and the lung lesions were diploid, but in the other patient both were found to be aneuploid at autopsy. No significant differences of S-phase fraction were observed between two different grade groups. There was no significant correlation among DNA ploidy, histologic grade, and the presence or absence of lung metastases. CONCLUSIONS: Based on this study, the DNA analysis has a limitation in predicting the biologic behavior of GCT.

Adolescent

Tandem repeat of the 23S and 5S ribosomal RNA genes in Borrelia burgdorferi, the etiological agent of Lyme disease.

The DNA fragments containing the rrl and rrf genes were subcloned from a EMBL3 recombinant phage of Borrelia burgdorferi strain B31 into pUC18 and were characterized by restriction analysis and Southern hybridization. A fine restriction map of the fragments was constructed and the organization of the genes was determined. The genomic hybridization using the gene probes from B. burgdorferi showed that there are two sets of rrl/rrf genes in that genome. The results also revealed the important fact that the gene sets are repeated directly by 3.2-kb long. This is the first report of this remarkable feature in the organization of the eubacterial rRNA genes.

Borrelia burgdorferi Group

Bone mineral density for patients with bone metastasis of prostate cancer: a preliminary report.

Patients with prostate cancer frequently have osseous metastases which are qualitatively assessed with bone scannings. We have studied the quantitative evaluation of skeletal diseases by measuring bone mineral density (BMD) in the lumbar vertebrae and radius. Forty-four patients with prostate cancer, nine with non-prostatic urogenital cancer and 90 controls were entered in this study. Among the patients with prostate cancer, the values of BMD in the lumbar vertebrae were significantly higher in patients with osseous metastasis than in those without metastasis, whereas the values of BMD in the radius were insignificantly different. Most of the patients with high levels of BMD in the lumbar vertebrae had osseous metastatic disease with no relationship between BMD in the lumbar vertebrae and the radius. The values of BMD in the lumbar vertebrae where hot spot scans were observed were related to X-ray findings. The alterations of BMD levels in the lumbar vertebrae were quantitatively evaluable as responses to androgen deprivation therapy. Measurement of BMD is useful for the accurate diagnosis of osteosclerotic lesions. BMD measurements of the lumbar vertebrae compared with those in the radius were variable in individuals.

Adenocarcinoma

Accumulation of 99mTc-HM-PAO in photon deficient areas in bone scan of bone metastasis from hepatocellular carcinoma.

To evaluate bone metastasis from hepatocellular carcinoma (HCC), both bone and 99mTc-HM-PAO scintigraphies were performed in six patients with clinically and pathologically confirmed HCC. Two patients had a bone scintigram which revealed abnormal accumulation in the skull base, pelvic bone and thoracic spine. The 99mTc-HM-PAO scans of both these patients also showed abnormal accumulation in the same sites. The bone scintigrams in one patient revealed not only abnormal accumulation in the ribs but also photon deficient areas in the sternum, thoracic spine and femur, while 99mTc-HM-PAO scans showed abnormal accumulation in all these sites. In three patients, bone scintigraphy revealed photon deficient areas in the ribs, pelvic bone and femur, and their 99mTc-HM-PAO scintigrams showed abnormal accumulation in the same sites. Thus, it was shown that, in the detection of bone metastasis from HCC by means of bone scintigraphy, it was necessary to pay attention to hot and cold lesions, and that a combination study with 99mTc-phosphorous compounds and 99mTc-HM-PAO was useful in evaluating these lesions.

Aged

FK506-induced kidney tubular cell injury.

Some renal changes associated with cyclosporine, such as tubular vacuolization and glomerular thrombosis, have also been reported with FK506. Furthermore, FK506 therapy is associated with a decrease in glomerular filtration rate and renal plasma flow and an increase in renal vascular resistance. We studied the in vitro tubular cell sensitivity to FK506 in comparison with CsA, the ultrastructural changes induced by FK506 and CsA, and the effect of both drugs on tubular cell growth in vitro. We also investigated whether FK506 and CsA induced endothelin-1 (ET-1) secretion of cultured tubular cells and whether this stimulatory effect coincided with a change in the endothelin systemic synthesis. Exposure of tubular cells to high concentrations of FK506 or CsA (10, 50, 100 microM) induced a time- and dose-dependent cell injury in vitro. The damage induced by FK506 and CsA was characterized by a direct cytotoxic effect on tubular cells, as expressed by release of 3H thymidine from prelabeled cells, N-acetyl-beta-D-glucosaminidase release, and cell detachment. Ultrastructural changes (vacuolizations, swelling, and mitochondrial enlargement) and inhibition of the growth of cultured tubular cells were also observed at high concentrations of FK506 and CsA. Low concentrations of FK506 and CsA (1, 0.1, 0.01, 0.001 microM) were not cytotoxic and induced only a minimal inhibitory effect on the growth of tubular cells in vitro. We demonstrated that FK506 (1, 0.1, 0.01 microM) time-dependently stimulated the secretion of endothelin by cultured tubular cells. CsA 10, 1, 0.1, 0.01 also exerted an enhancing effect on ET-1 secretion in cultured tubular cells. We observed that the concentration of CsA that induced the most important enhancing effect was 10 or 100 times higher than that required for FK506 to observe the same effect. The concentrations of FK506 or CsA that induced ET-1 secretion were not cytolytic for tubular cells in vitro. FK506- or CsA-treated rats showed an increase in serum level of ET-1 in comparison with the control. Through the stimulatory effect on endothelin secretion by tubular cells, FK506 and CsA may induce a perturbation of renal hemodynamics. Concentrations of FK506 and CsA, higher than established serum levels but close to those reached in tissues, are cytotoxic for tubular cells and induced ultrastructural changes and a significant delayed regeneration.

Acetylglucosaminidase

The 23S/5S ribosomal RNA genes (rrl/rrf) are separate from the 16S ribosomal RNA gene (rrs) in Borrelia burgdorferi, the aetiological agent of Lyme disease.

DNA fragments containing the rRNA genes for Borrelia burgdorferi strain B31 were cloned in bacteriophage lambda EMBL3. A restriction map of the fragments was constructed and the organization of the rRNA genes was determined by Southern hybridization. One genomic DNA fragment contained a single copy of the rrs sequence and another cloned fragment contained both rrl and rrf sequences. The results revealed that the rrs gene is located separately from the set of rrl/rrf genes, suggesting that these rRNA genes are expressed independently in B. burgdorferi.

Blotting, Southern

Organization of the ribosomal RNA genes in Treponema phagedenis and Treponema pallidum.

The genomic DNA fragment which contains ribosomal RNA (rRNA) genes for Treponema phagedenis was cloned into bacteriophage vector lambda EMBL3. A restriction map of the fragment was constructed and the organization of the rRNA genes was determined. The fragment contained at least one copy of the 16S, 23S and 5S sequences and the genes are arranged in the order 16S-23S-5S. Southern hybridization using radiolabeled rRNA gene probes to genomic DNA from T. phagedenis strain Reiter and T. pallidum strain Nichols showed that these organisms have two radioactive fragments which hybridize to the probes in their genome. These results suggest that both pathogenic and non-pathogenic strains of Treponema may carry at least two sets of rRNA genes on their chromosomes.

Cloning, Molecular

Atypical adenoma of the thyroid. A clinicopathologic and flow cytometric DNA study in comparison with other follicular neoplasms.

A clinicopathologic and DNA flow cytometric study was performed on seven patients (three males, four females) with atypical adenoma of the thyroid gland, using formalin-fixed paraffin-embedded tissues. The results were compared with those of 30 follicular adenomas and 13 follicular carcinomas. The patients ranged in age from 32 to 74 years (mean; 55.8 years), and the mean follow-up period was 11.0 years. All patients except two who died of other diseases were free of thyroid disease after initial surgery. It showed that there was no evidence of clinical cancer in this follow-up study of patients operated on for atypical adenomas. Four of the atypical adenomas were diploid, two were aneuploid, and one was tetraploid. Twenty-seven of the 30 follicular adenomas were diploid. Three patients with aneuploid follicular adenoma were free of disease. Of the 13 follicular carcinomas with a mean follow-up period of 6.9 years, five were diploid, seven were aneuploid, and one was tetraploid. Two patients with aneuploid follicular carcinoma and one with diploid carcinoma developed lung metastases, and one patient each with diploid and aneuploid follicular carcinoma died of disease. There was no significant correlation between histologic features, ploidy status and prognosis among follicular carcinomas. The results of this study suggest that DNA flow cytometric analysis is not a useful tool for predicting the clinical behavior of follicular neoplasms.

Adenoma

Glomerular actions of a free radical-generated novel prostaglandin, 8-epi-prostaglandin F2 alpha, in the rat. Evidence for interaction with thromboxane A2 receptors.

8-epi-prostaglandin F2 alpha (8-epi-PGF2 alpha) and related compounds are novel prostanoid produced by a noncyclooxygenase mechanism involving lipid peroxidation. Renal ischemia-reperfusion injury increased urinary excretion of these compounds by 300% over baseline level. Intrarenal arterial infusion at 0.5, 1, and 2 micrograms/kg per min induced dose-dependent reductions in glomerular filtration rate (GFR) and renal plasma flow, with renal function ceasing at the highest dose. Micropuncture measurements (0.5 microgram/kg per min) revealed a predominant increase in afferent resistance, resulting in a decrease in transcapillary hydraulic pressure difference, and leading to reductions in single nephron GFR and plasma flow. These changes were completely abolished or reversed by a TxA2 receptor antagonist, SQ 29,548. Competitive radioligand binding studies demonstrated that 8-epi-PGF2 alpha is a potent competitor for [3H]SQ 29,548 binding to rat renal arterial smooth muscle cells (RASM) in culture. Furthermore, addition of 8-epi-PGF2 alpha to RASM or isolated glomeruli was not associated with stimulation of arachidonate cyclooxygenase products. Therefore, 8-epi-PGF2 alpha is a potent preglomerular vasoconstrictor acting principally through TxA2 receptor activation. These findings may explain, in part, the beneficial effects of antioxidant therapy and TxA2 antagonism observed in numerous models of renal injury induced by lipid peroxidation.

Animals

Comparison of flanking regions of the 5S ribosomal ribonucleic acid genes in Leptospira biflexa and Leptospira interrogans.

One of the genes encoding the 5S ribosomal ribonucleic acid (rRNA) for the Leptospira biflexa strain Patoc I was isolated and sequenced. The physical maps of the 5S rRNA genes in Leptospira were constructed. The strains of Leptospira biflexa had two genes on their chromosome; these two 5S rRNA genes were located several kb apart and sequences flanking these genes were divergent. In contrast to saprophytic leptospires, maps in parasitic leptospires that had only one gene for 5S rRNA on their genome were highly conserved and the physical maps of the genes in almost all strains were similar.

Base Sequence

[Phase III trial of 99mTc-rhenium colloid for lymphoscintigraphy].

A multicenter study was carried out on 191 patients (196 examinations) with lymphatic system disorders to evaluate the efficacy and safety of 99mTc-rhenium Colloid, a tracer for lymphoscintigraphy (TCK-17). Local pain and swelling occurred at the site of injection in 79.6% and 5.1% of patients, respectively, and 2 patients experienced mild fever. The accuracy was calculated on the basis of the results obtained by other diagnostic methods. Lymphoscintigraphy using TCK-17 was sensitive diagnostic procedures, but low specificity. The efficacy was classified into five grades: "Excellent", "Good", "Moderate", "Equivocal", and "Poor". 67.3% of all examination were evaluated as "Excellent" or "Good". This study revealed TCK-17 was a useful radiopharmaceutical for lymphoscintigraphy because of its safety and effectiveness.

Adult

[A fundamental study of 99mTc-ethyl cysteinate dimer (99mTc-ECD)].

In vitro fundamental studies of 99mTc-ethyl cysteinate dimer (99mTc-ECD), a newly developed radiopharmaceutical for cerebral perfusion scintigraphy, were performed, and were compared with those of 99mTc-HM-PAO, which has been widely applied to clinical use. 99mTc-HM-PAO should not only be injected soon after reconstitution, but also be avoided mixture with saline or drugs such as Diamox, because of its radiochemical purity decreases with the passage of time and the addition of saline and Diamox. On the other hand, the radiochemical purity of 99mTc-ECD reached a constant level of 94-96% at 30 to 180 min after reconstitution, and it remained stable against the addition of saline and Diamox. Therefore, it was suggested that 99mTc-ECD could be injected from an already saline-infused venous-line or mixed with Diamox. However, early injection after reconstitution should be avoided, because 99mTc-ECD did not become stable enough to maintain sufficient radiochemical purity until 30 min after reconstitution. The present studies are suggested that the red blood cell component is largely related to the metabolism of 99mTc-ECD in blood. Furthermore, from study of the metabolic rate using rabbit blood, it was suggested that a species difference existed in esterase activity in blood.

Animals

[A case of gallbladder cancer with marked response to EAP treatment].

A case of unresectable gall bladder cancer due to multiple metastasis was dealt with EAP (VP-16, ADM, CDDP) treatment. The case was a 49-year-old male who was admitted to the hospital with a 3 cm sized umbilical tumor. After 2 cycles of the treatment, the tumor size significantly decreased and the symptoms diminished. In parallel, the symptoms from bone metastasis and dissemination also disappeared. As for tumor markers, CA 19-9 indicated 5151 U/ml, fell down to the normal range after the treatment. In this case, the efficacy of EAP treatment to the primary gall bladder carcinoma was undetectable, because of the difficulty of elucidation of the primary site by diagnostic images. However, all data suggested that EAP treatment was effective to the advanced unresectable gallbladder cancer, at least to its metastatic sites.

Adenocarcinoma

Signal transduction mechanism of interleukin 6 in cultured rat mesangial cells.

Interleukin 6 (IL-6) is one of the potent autocrine growth factors for mesangial cells. We investigated the signal transduction mechanism of IL-6 in cultured rat mesangial cells. IL-6 induced a transient increase of inositol 1,4,5-trisphosphate (Ins 1,4,5-P3) followed by a transient and sustained increase of intracellular calcium concentration, suggesting that IL-6 stimulates phosphoinositide turnover. IL-6 also stimulated prostaglandin E2 (PGE2) production. The IL-6-concentration dependency in PGE2 production was similar to that in Ins 1,4,5-P3 production. We concluded that the action of IL-6 on mesangial cells is exerted at least partially through the enhancement of phosphoinositide turnover and PGE2 production.

Animals

HLA-DQ-restricted CD4+ T cells specific to streptococcal antigen present in low but not in high responders.

We have found that the low immune response to streptococcal cell wall Ag (SCW) was inherited as a dominant trait and was linked to HLA, as deduced from family analysis. In the present report, HLA class II alleles of healthy donors were determined by serology and DNA typing to identify the HLA alleles controlling low or high immune responses to SCW. HLA-DR2-DQA1*0102-DQB1*0602(DQw6)-Dw2 haplotype or HLA-DR2-DQA1*0103-DQB1*0601(DQw6)-DW12 haplotype was increased in frequency in the low responders and the frequency of HLA-DR4-DRw53-DQA1*0301-DQB1*0401(DQw4)-Dw15 haplotype or HLA-DR9-DRw53-DQA1*0301-DQB1*0303(DQw3)-Dw23 haplotype was increased in the high responders to SCW. Homozygotes of either DQA1*0102 or DQA1*0103 exhibited a low responsiveness to SCW and those of DQA1*0301 were high responders. The heterozygotes of DQA1*0102 or 0103 and DQA1*0301 showed a low response to SCW, thereby confirming that the HLA-linked gene controls the low response to SCW, as a dominant trait. Using mouse L cell transfectants expressing a single class II molecule as the APC, we found that DQw6(DQA1*0103 DQB1*0601) from the low responder haplotype (DR2-DQA1*0103-DQB1*0601(DQw6)-Dw12) activated SCW-specific T cell lines whereas DQw4(DQA1*0301 DQB1*0401) from the high responder haplotype (DR4-DRw53-DQA1*0301-DQB1*0401(DQw4)-Dw15) did not activate T cell lines specific to SCW. However, DR4 and DR2 presented SCW to CD4+ T cells in both the high and low responders to SCW, hence the DR molecule even from the low responder haplotype functions as an restriction molecule in the low responders. Putative mechanisms linked to the association between the existence of DQ-restricted CD4+ T cells specific to SCW, and low responsiveness to SCW are discussed.

Bacterial Proteins

Case report 679. Central low-grade osteosarcoma with foci of dedifferentiation.

An infrequently encountered case of well-differentiated CLOS containing dedifferentiated tumor foci in a 28-year-old Japanese man is reported. The patient died from widespread metastases with a microscopic appearance identical to the anaplastic dedifferentiated lesions of the original tumor. This is, to our knowledge, the first report of a case of CLOS with dedifferentiation at the time of initial surgery. The literature concerning CLOS was reviewed and the diagnostic features of CLOS discussed.

Adult