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Biomedical subjects

S Minoshima

Publications and source records attributed to S Minoshima.

At least 217 records · Page 12Linked to original sources

Shotgun polymerase chain reaction: construction of clone libraries specific to a NotI fragment of flow-sorted human chromosome 22.

We have developed the "shotgun polymerase chain reaction," a method for obtaining a large number of DNA markers specific to a giant DNA fragment, which facilitates analysis of a particular chromosomal region. We applied this method to a giant NotI fragment which carries the immunoglobulin lambda constant region on chromosome 22. NotI digests of chromosome 22 flow-sorted from human B-lymphoblastoid cell line GM130B were size fractionated by pulsed-field gel electrophoresis. Preliminary Southern hybridization analysis revealed that the immunoglobulin lambda constant region was conveyed on 1.4- and 1.3-Mb NotI fragments in this cell line. The agarose gel corresponding to 1.2 to 1.5 Mb in size was excised into slices and subjected to polymerase chain reaction to identify gel slices containing NotI fragments carrying Ke-Oz+, a subtype of the immunoglobulin lambda constant region. From the NotI fragment thus identified, a large number of small DNA segments were amplified through the ligation-mediated random polymerase chain reaction method. The amplified products were cloned and analyzed for chromosomal origin and localization to particular NotI fragments. Seven of eighteen clones originated from the 1.4-Mb NotI fragment of chromosome 22 in GM130B cells, which appears to be exactly the same as detected by a probe for the immunoglobulin lambda constant region.

Base Sequence↗

Molecular cloning and chromosomal mapping of a cDNA encoding human 80K-L protein: major substrate for protein kinase C.

We have isolated and sequenced complementary DNA (cDNA) for the human 80K-L protein, a major substrate for protein kinase C and the human homologue of an 80- to 87-kDa bovine protein named MARCKS (myristoylated alanine-rich C kinase substrate). The human 80K-L cDNA encodes a protein of 332 amino acids with a calculated molecular weight of 31,534. Homology comparisons of the nucleotide sequences of the cDNAs indicated that their 3'-untranslated regions are more homologous than the coding regions. Spot blot hybridization using flow-sorted human chromosomes indicated that the gene encoding the 80K-L protein, designated MACS, maps to the q15----qter region of human chromosome 6, and it also suggested that a genomic region with a sequence homologous to the 3'-untranslated region of the 80K-L mRNA exists on chromosome 21.

Amino Acid Sequence↗

Human long-chain acyl-CoA synthetase: structure and chromosomal location.

A complementary DNA clone encoding the entire human long-chain acyl-CoA synthetase was isolated and the total 698-amino acid sequence was deduced. The amino acid sequence of human long-chain acyl-CoA synthetase shows 84.9% identity to that of rat long-chain acyl-CoA synthetase. The nucleotide sequences of the protein coding regions between human and rat long-chain acyl-CoA synthetase mRNAs are highly conserved (85.6%), whereas those of the 3' untranslated regions are less conserved (72%). The location of the human long-chain acyl-CoA synthetase gene was identified on chromosome 4 by spot hybridization of flow-sorted chromosomes. Computer-assisted homology search revealed a significant similarity of the enzyme with the enzymes of the luciferase family. Based on this similarity, the structure of human long-chain acyl-CoA synthetase can be divided into five domains: the N-terminus, two domains similar to those in enzymes of the luciferase family, a long gap region between the similar domains and the C-terminus.

Amino Acid Sequence↗

A case of Goodpasture's syndrome associated with anti-myeloperoxidase antibodies.

A case of Goodpasture's syndrome with anti-myeloperoxidase (MPO) antibodies is reported. Histological examination revealed crescentic glomerulonephritis and alveolar hemorrhage with linear deposition of IgG along the glomerular capillary walls and alveolar capillary walls by immunofluorescence microscopy. Not only anti-glomerular basement membrane (GBM) antibodies but also anti-MPO antibodies, an anti-neutrophil cytoplasmic antibody, were simultaneously detected in the serum. Although it is generally accepted that crescentic glomerulonephritis in Goodpasture's syndrome is mediated by anti-GBM antibodies, this case suggested that anti-MPO antibodies might also participate in the pathogenesis of crescentic glomerulonephritis and probably alveolar hemorrhage of Goodpasture's syndrome, especially with vasculitis.

Anti-Glomerular Basement Membrane Disease↗

[A case of microscopic polyarteritis nodosa associated with myeloperoxidase-antineutrophil cytoplasmic autoantibodies (MPO-ANCA)].

We report a case of microscopic polyarteritis nodosa associated with myeloperoxidase-antineutrophil cytoplasmic autoantibodies (MPO-ANCA). A 38 year-old female was admitted to our hospital, because of proteinuria, recurrent pyrexia, polyarthralgia, abdominal pain and purpura. She had a history of severe pulmonary hemorrhage and 4 kg weight loss for 8 months. On admission perinuclear ANCA without cytoplasmic ANCA was detected by indirect immunofluorescence assay and MPO-ANCA was detected by enzyme linked immunosorbent assay. But anti-nuclear antibodies, immune complexes and anti-glomerular basement membrane antibodies were not detected. Renal biopsy showed necrotizing crescentic glomerulonephritis without immune deposits. Skin biopsy revealed leukocytoclastic vasculitis. Diagnosis of microscopic polyarteritis nodosa was made by these clinical and histological evidence of vasculitis. As renal failure progressed after admission, corticosteroid and cyclophosphamide administration were started. Renal function and other symptoms improved paralleled with decreased MPO-ANCA titer to normal values. It is suggested that MPO-ANCA may be closely related to the pathogenesis of microscopic polyarteritis nodosa and it may be a good serological marker for diagnosis and disease activity of this disease.

Adult↗

[Discrepancy between 123I-IMP SPECT and X-ray CT in chronic cerebrovascular disease].

The discrepancy between the low accumulation area (LAA) seen on 123I-IMP SPECT and the low density area (LDA) on X-ray CT was evaluated in 76 cases with chronic cerebrovascular disease. In 19 patients, LAA was larger than LDA beyond the vascular territory. Brain angiogram showed either obstruction or stenosis of the major arteries in as many as 59 percentages of this discrepancy group. In many patients of this group, cortical neurological symptoms were observed. In 37 patients in which the size of LAA was equal to that of LDA, abnormality of the major arteries was observed in only 7 percentages. These results indicated that the discrepancy mainly reflected the infarction due to irreversible ischemic change of perforate artery with the cortical low perfusion caused by the major artery disease. In some patients, the discrepancy was believed to show the remote effect caused by neurological pathway disturbance.

Adult↗

[Visualization of the female breast in bone scintigraphy].

Breast cancer incidence rates in Japan have more than doubled in the decade between 1975 and 1985 due to a well-established and affluent Western life-style. One hundred patients having breast cancer, 107 patients having mastectomies, and 243 patients having no breast cancer were reviewed for whole body and spot views of bone scintigraphy with 99mTc-MDP. The frequency of positive breast uptake in patients with no breast cancer showed a maximum in those patients between the ages of 30 and 40, with a progressive rise and fall before and after the peak, respectively. Patients between the ages of 50 and 60 with breast cancer showed a maximum uptake of 83%, while those patients between the ages of 70 and 80 with breast cancer showed an uptake higher than 50%. There was a markedly higher breast uptake ratio in the breast cancer group compared to the breast cancer free group. Although this scanning agent was of no value in recognition of tumor localization from breast uptake, we have to suspect a high incidence of cancer existence whenever there was unilateral increased breast activity. Even though the mechanism of 99mTc-MDP uptake by breast tissue is unknown, breast concentration of 99mTc-MDP appears to be influenced by hormones, especially estrogen.

Adolescent↗

An automated method for rotational correction and centering of three-dimensional functional brain images.

The display and analysis of functional brain images often benefit from head rotational correction and centering. An automated method was developed to align brain PET images into a standard three-dimensional orientation. The algorithm performs transverse and coronal rotational correction as well as centering of a brain image set. Optimal rotational correction and centering are determined by maximizing a bilateral hemispheric similarity index, the stochastic sign change criterion. Testing of this algorithm on simulated symmetrical brain image sets showed errors less than 1.0 degree and 0.5 pixels for rotational correction and centering, respectively. With actual PET data, the algorithm results correlated well with those obtained by visual inspection. Testing on asymmetrical brain image sets with simulated lesions indicated that performance of the algorithm is not sensitive to focal asymmetries. This automated method provides objective, reproducible image alignment into a standard orientation and facilitates subsequent data analysis techniques for functional brain images.

Algorithms↗

Flow cytometric analysis of the expression of 9A3 antigen, E-cadherin and EGF receptor in TMK-1 stomach cancer cells.

We have previously produced a monoclonal antibody (MAb) against TMK-1 cells, a poorly differentiated stomach-cancer cell line, and designated the antigen as 9A3. Immunohistochemical analysis of various tumor tissues revealed that the expression of 9A3 antigen is specific for human adenocarcinoma. In the present study, we analyzed the expression patterns of 9A3 antigen on TMK-1 cells and compared them with the expression patterns of E-cadherin and epidermal-growth-factor (EGF) receptors, using immunofluorescent microscopy and flow cytometry. The flow cytometric analysis revealed that the TMK-1 cell population consists of 9A3-positive cells and 9A3-negative cells. The isolated 9A3-negative cell population generates 9A3-positive cells as they divide, whereas the isolated 9A3-positive cell population does not generate 9A3-negative cells. The 9A3-positive cells grow as tight colonies, whereas the 9A3-negative cell form loose colonies in soft agar culture. E-cadherin was expressed only in the 9A3-positive TMK-1 cells and not in the 9A3-negative cells. In contrast, EGF receptor was expressed in both cell populations. The 9A3 antigen was localized mainly at the cell-cell adhesion sites where E-cadherin was apparently condensed.

Adenocarcinoma↗

Structure and chromosome assignment of human S100 alpha and beta subunit genes.

The human S-100 alpha and beta subunit genes were isolated from human genomic DNA library. The restriction endonuclease mapping and DNA sequencing analysis revealed that both subunit genes consisted of three exons and two introns. Two Ca2(+)-binding domains were independently encoded by exon 2 and exon 3. Spot-blot hybridization analysis with flow-sorted chromosomes showed that human alpha and beta subunit genes were assigned to chromosome 1 and chromosome 21, respectively.

Amino Acid Sequence↗

[Magnetic resonance imaging of small nasopharyngeal tumors].

Magnetic resonance (MR) images of 9 patients with a small nasopharyngeal tumor were examined, and among them three cases were evaluated with Gd-DTPA enhancement technique. All cases showed no asymmetry of parapharyngeal space, but they presented neck lymph nodes swelling. These cases were proved malignant by surgical procedures and the blind biopsy. MR findings of levator and tensor veli palatini muscles and retropharyngeal lymph nodes were retrospectively evaluated in correlation with endoscopic examination and biopsy. In six cases among 9 patients, MR images demonstrated tumors on the lateral wall of nasopharynx. Infiltration of levator veli palatini muscle was an especially important earlier sign of nasopharyngeal tumor invasion because the levator veli palatini muscle belongs to the intrapharyngeal muscle group. Endoscopic examination could not reveal any abnormal lesion in three cases among these 6 cases. No definite signal intensity difference between tumor and pharyngeal mucosa was shown on both T1 and T2 weighted images. But the normal pharyngeal mucosa had demonstrated higher intensity than tumor with Gd-DTPA administration. So the enhanced nasopharyngeal MR imaging made the clear intensity difference between tumor and muscle tissue. It was the conclusion that MR imaging was superior to X-CT in detecting small nasopharyngeal tumor.

Adult↗

Technetium-99m HMPAO labeled leukocytes in inflammation imaging.

Technetium-99m-HMPAO (Tc-99m-HMPAO) labeled leukocyte imaging was carried out in 19 patients at 3-5 hr after reinjection. There were no side effects noted. Tc-99m leukocyte images showed gall bladder, colon, kidney, and urinary bladder activity in normal distribution as a result of excretion of the eluted Tc-99m complex. They yielded a sensitivity of 93%, a specificity of 100% and an accuracy of 95%. They were correctly positive in 14 out of 19 cases. But one false negative case was seen in a patient with pyonephrosis showing a lack of renal function with decreased renal blood flow. It was concluded that they have some advantages over In-111 leukocyte images, but we have to consider the fact that the ureteral obstruction or the lack of renal function with decreased renal blood flow may result in a false positive or a false negative case.

Adult↗

Regional localization of the hepatocyte growth factor (HGF) gene to human chromosome 7 band q21.1.

Hepatocyte growth factor (HGF) is a polypeptide involved in liver regeneration. Its amino acid sequence and gene structure are similar to those of coagulation-related serine proteases. We have used a cDNA clone of HGF and flow-sorted human chromosomes to assign this gene to chromosome 7. Fluorescence in situ hybridization of the HGF genomic clones to human metaphase chromosome spreads showed the localization of this gene to 7q21. Estimation of fluorescent signals relative to arbitrary reference points (ARPs) allowed further localization to 7q21.1.

Cell Fractionation↗

Localization of the human gene for 230-kDal bullous pemphigoid autoantigen (BPAG1) to chromosome 6pter----q15.

Chromosome mapping of the human gene encoding the 230-kDal autoantigen of an autoimmune skin disease, bullous pemphigoid, was performed using flow-sorted human chromosomes of cells of normal karyotype and cells carrying a reciprocal translocation t(6;16)(q15;q24). The cDNA of the autoantigen hybridized with intact chromosome 6 and translocation chromosome 6p- (6pter----q15::16q24----qter). The gene (BPA230) was located to the chromosome region 6pter----q15.

Animals↗

Assignment of a human serum glycoprotein SP-40,40 gene (CLI) to chromosome 8.

SP-40,40 is a serum glycoprotein consisting of two different subunits (alpha and beta) assembled into a dimer by disulfide bonds. Northern blot hybridization, using total RNA from several cell lines, showed that SP-40,40 is expressed in glioblastoma and testicular tumor cells, as well as hepatoma cells. Spot blot hybridization of flow-sorted human chromosomes, using a SP-40,40 cDNA fragment as a probe, localized the gene for SP-40,40 to human chromosome 8. This gene has been given the designation CLI, for complement lysis inhibitor, by the Human Gene Nomenclature Committee.

Blood Proteins↗

[A case of anti-myeloperoxidase antibodies-associated idiopathic crescentic glomerulonephritis with pulmonary hemorrhage].

We report a case of idiopathic crescentic glomerulonephritis with pulmonary hemorrhage associated with anti-myeloperoxidase antibodies (anti-MPO ab). A 74 year-old female was admitted to our hospital because of rapidly progressive glomerulonephritic syndrome and dyspnea with bloody sputum. On admission anti-MPO ab, one of anti-neutrophil cytoplasmic antibodies, were detected but anti-GBM antibodies and immune complexes were not detected. Renal biopsy showed crescentic glomerulonephritis and lung biopsy showed massive alveolar hemorrhage. Both tissue had pauci-immune deposit by immunofluorescence microscopy. Hemodialysis and steroid administration were started. Pulmonary hemorrhage was improved remarkably, but renal failure progressed rapidly to end stage kidney, then hemodialysis was continued. Although subsequent 3 years uneventful maintenance hemodialysis had been performed, she admitted to our hospital again because of progressive dyspnea with hemoptysis after upper respiratory tract infection. On admission anti-MPO ab were detected again and steroid administration was started. Pulmonary hemorrhage was improved with decreased anti-MPO ab titer. While tapering the dosis of steroid, anti-MPO ab again increased and pulmonary hemorrhage recurred. Although pulse methylprednisolone therapy and plasma exchange were performed, respiratory failure progressed rapidly and she died of sepsis. Postmortem examination showed no evidence of systemic vasculitis. In this case, titer of anti-MPO ab was associated with not only idiopathic crescentic glomerulonephritis but also with pulmonary hemorrhage. We tried to detect enzymatically active MPO in serum. Titer of serum MPO was also associated with disease activity and anti-MPO ab. It is suggested that both anti-MPO ab and serum MPO are closely related to the pathogenesis of idiopathic crescentic glomerulonephritis and pulmonary hemorrhage.

Aged↗

[Blood flow measurement of brain tumor by 123I-IMP using 3-heads rotating gamma camera SPECT].

Using 3-heads rotating gamma camera SPECT system, regional blood flow was measured from the super-early image obtained 4 to 6 minutes after the intravenous injection of 123I-IMP on 11 patients with various brain tumors. Two cases of olfactory groove meningioma and malignant astrocytoma showed significantly high 123I-IMP uptake by the lesion visualized by MRI compared to the uptake by normal brain cortex on super-early image but 123I-IMP uptake by the tumor decreased on early image. In both cases, blood flow of the tumor measured from super-early image showed values much higher than those of normal brain cortex. It was thought that usual correction method based on early image did not correctly represent the blood flow of the lesion but that the present method based on super-early image could show the increase of blood flow of the tumor.

Adult↗

The use of FDG-PET in the detection and management of malignant lymphoma: correlation of uptake with prognosis.

Twenty-one patients with untreated malignant lymphoma in the head and neck region were evaluated with positron emission tomography (PET) using fluorine-18-fluorodeoxyglucose (FDG) and gallium-67 SPECT imaging. Tumor-to-normal soft-tissue contrast ratios (TCRs) obtained 60 min after injection of FDG were higher than 2.6, and all malignant lymphomas were clearly visualized. In patients with poor prognosis, higher TCRs and glucose utilization rates (GURs) were observed, whereas low TCR and GUR were shown in a patient with low-grade malignancy. In comparison with 67Ga scintigraphy, patients with high TCRs and GURs were likely to show increased accumulation of gallium-67, but accumulation of gallium-67 was not increased as much as FDG in poor prognostic patients. FDG-PET may be useful in the detection and management of malignant lymphoma.

Adult↗