[A case of anti-lipid syndrome manifesting portal hypertension and intra-hepatic nodular hyperplasia].
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Abortion, Habitual↗
Biomedical subjects
S Minoshima
Publications and source records attributed to S Minoshima.
Isolation of giant DNA fragments from flow-sorted human chromosomes.
We have established a method using a conventional cell sorter equipped with a single argon laser to sort intact human chromosomes that can be used as a source for the production of giant DNA fragments. Various improvements were made to both the equipment and sorting method to enhance the sorting resolution and avoid destruction of chromosomal DNA. Using this improved method chromosomes 21 and 22 were sorted from the B-lymphoblastoid line GM00130B, digested with the rare cutting restriction endonuclease NotI, and analyzed by pulsed field gel electrophoresis followed by Southern hybridization using the Alu repetitive sequence as a probe. More than 25 discrete NotI giant DNA fragments ranging from 50 kb to longer than 2.5 Mb were separated and the size distribution pattern was unique for each chromosome, indicating successful sorting of intact chromosomes. The cumulative size of these Alu-positive NotI DNA fragments were 22.7 Mb and 25.5 Mb for chromosomes 21 and 22, respectively. These values are 47% and 49% of the estimated size of chromosomes 21 (48 Mb) and 22 (52 Mb).
Cell Line, Transformed↗
Four genes for the calpain family locate on four distinct human chromosomes.
Calcium dependent proteases (calpains, CAPNs, E.C.3.4.22.17) constitute a family of proteins which share a homologous cysteine-protease domain (large subunits, L1, L2, and L3) and an E-F hand Ca2(+)-binding domain (L1, L2, L3, and small subunit, S). We have mapped the genes for four calpain proteins (L1, L2, L3, and S) on four distinct human chromosomes by a combination of spot-blot hybridization to flow-sorted chromosomes and Southern hybridization of DNAs from a human x mouse hybrid cell panel. The genes for calpain L1 (CAPN1, large subunit of calpain I), L2 (CAPN2, large subunit of calpain II), L3 (CAPN3, a protein related to the large subunits), and S (CAPN4, a small subunit common to calpains I and II) were assigned to human chromosomes 11, 1, 15, and 19, respectively.
Calpain↗
Assignment of the human calpastatin gene (CAST) to chromosome 5 at region q14----q22.
The human calpastatin gene (CAST) was assigned to chromosome 5 by spot-blot hybridization analysis with flow-sorted chromosomes, and it was further sublocalized to bands 5q14----q22 using in situ hybridization to metaphase chromosomes.
Calcium-Binding Proteins↗
[Evaluation of image quality and lesion detectability on I-123 IMP brain SPECT].
Influence of image quality on lesion detectability in I-123 IMP brain SPECT was discussed in forty-two cases with various cerebrovascular disorders. Fifty nine lesions, which were considered to show decreased uptake of I-123 IMP, were analyzed in this study. In all cases, SPECT examinations were started within 30 minutes after intravenous administration of 3 mCi of I-123 IMP. To determine the lesion detectability, fourteen radiologists from nine hospitals were gathered and film reading was carried out. To classify the image quality into valuable or not, findings of five points on SPECT image were visually evaluated; 1) uniformity of cortex accumulation, 2) separation of bilateral visual cortices, 3) density of white matter, 4) separation between basal ganglion and thalamus, 5) density of basal ganglion and thalamus. Mean lesion detections were 10.3/14 for the lesions of cerebral cortex, 9 for cerebellum, 3.58 for white matter and 3.22 for basal ganglion and thalamus, respectively. The detectability was certainly influenced by the image quality in the lesion of basal ganglion and thalamus, white matter and single lobe of cerebral cortex. We concluded that detection of small lesions and deep lesions with I-123 IMP SPECT was dependent on the image quality and the proposed classification of I-123 IMP SPECT image was useful for evaluating image quality from the points of view of the lesion detectability.
Adolescent↗
[A case of subacute cerebral infarction demonstrating hyperemia and crossed cerebellar diaschisis in I-123 IMP SPECT].
SPECT with N-isopropyl-p-[I-123]iodoamphetamine were performed in a 81-year-old man with cerebral infarction. In the subacute phase, the radioactivity was increased in the infarct area where fogging effect and remarkable contrast enhancement was demonstrated in X-ray CT. The contralateral cerebellar hemisphere showed reduced activity due to crossed cerebellar diaschisis (CCD). In the chronic phase, the decrement of the activity in the infarct was observed. Increased activity in the subacute phase was thought to reflect the hyperemia in the infarct, while CCD suggested the decreased metabolic activity in the lesion. The coexistence of the hyperemia and the CCD indicates flow and metabolic uncoupling, which means "luxury perfusion". This case was also thought to demonstrate atypical findings of CCD in SPECT imaging.
Aged↗
[A case of pulmonary embolism diagnosed by 111In labeled platelet scintigraphy].
Although the role of 111In platelet scintigraphy is established in the diagnosis of deep vein thrombosis, its indication in pulmonary embolism remains uncertain. We performed platelet scintigraphy in a patient with recurrent pulmonary thromboembolism, which revealed abnormal accumulation in the left hilar region. Pulmonary perfusion scintigraphy performed at the same time did not show perfusion defect. However, pulmonary angiography and thromboembolectomy demonstrated recurrent embolus in the hilus. This suggested that platelet aggregation in the left hilus shown by platelet scintigraphy was indeed related with recurrence of pulmonary embolism, and platelet scintigraphy is useful in detecting new development of thrombus in patient with recurrent pulmonary thromboembolism.
Blood Platelets↗
[Evaluation of pontine visualization with single photon emission computed tomography using N-isopropyl-p-(I-123)iodoamphetamine in normal and pathological cases].
Pontine visualization with single photon emission computed tomography (SPECT) using N-isopropyl-p-[I-123]iodoamphetamine (I-123 IMP) was evaluated in 99 cases including 46 cases of brain infarction, 25 of other cerebrovascular disorders and 28 of brain tumor and in 19 normal subjects. In each case, SPECT using a rotating gamma camera was started at 30 minutes after intravenous administration of 111 MBq of I-123 IMP. Transverse images were reconstructed parallel to the cantho-meatal line and displayed on films. Pontine visualization was visually classified into three grades: high radioactive accumulation confined to the pons with clear contour (Grade 3), faint accumulation with indistinct contour in the pontine region (Grade 1), the pontine accumulation was identified but its activity was less significant (Grade 2). The results showed that grade of the pontine accumulation was lower in pathological cases compared to normal subjects. Six cases with brain stem lesion demonstrated pontine accumulation of grade 1 in 4 case and of grade 2 in 2 cases. Lower grade was also revealed in 29 cases with crossed cerebellar diaschisis (CCD). No significant correlation was observed between classified grade of pontine accumulation and maximum counts of reconstructed images. We concluded that SPECT could detect brain stem lesions and that lower accumulation in pons with CCD positive cases was thought to partially reflect the reduced blood flow and metabolism in the pontine nuclei.
Adolescent↗
A human muscle adenine nucleotide translocator gene has four exons, is located on chromosome 4, and is differentially expressed.
The human heart-skeletal muscle adenine nucleotide translocator (ANT1) gene was isolated and sequenced. It spans 5.8 kilobases and contains four exons. The 5'-nontranscribed region contains typical CCAAT and TATA sequences, a 22-nucleotide pair inverted repeat and a 13-nucleotide pair sequence homologous to a similar region in the ATP synthase beta subunit gene. The region surrounding the first exon and intron is G+C-region surrounding the first exon and intron is G+C-rich, and the intron contains three Sp1 binding motifs. ANT1 was assigned to chromosome 4 using both flow-sorted chromosomes and segregating human-mouse hybrid cells. Additional ANT sequences were found on at least two other chromosomes. ANT1 transcripts were present at high levels in human heart and skeletal muscle but were almost undetectable in liver, kidney, and brain. By contrast, fibroblast ANT (ANT2) mRNAs were present in all five tissues. The unique nature and arrangement of the ANT1 transcriptional control elements may account for this differential expression.
Adult↗
Selective killing of squamous carcinoma cells by an immunotoxin that recognizes the EGF receptor.
We have conjugated a murine monoclonal antibody (B4G7) against the human epidermal growth factor (EGF) receptor to gelonin, a 60S ribosome inactivating protein, via N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP) and 2-iminothiolane. The B4G7-gelonin conjugate bound to the cell surface in proportion to the number of EGF receptors and competed with B4G7 antibody for binding to EGF receptors. The conjugate killed EGF receptor-hyperproducing squamous carcinoma cells (A431, NA, Ca9-22, TE5), and to some extent, human fibroblasts (HFO). It did not kill EGF receptor-deficient small-cell lung cancer cells (H69) and mouse fibroblasts (Swiss/3T3). Free B4G7, gelonin or a mixture of B4G7 and gelonin did not kill A431 cells. The number of EGF receptors was correlated to cytotoxicity at 10(-8) M of the conjugate, and the data were fitted to the regression equation: y = -35.83 log x +233.4 (correlation coefficient = -0.9995). These results suggest that the B4G7-gelonin conjugate may be a useful weapon for targeting therapy to squamous-cell carcinomas.
Animals↗
Complete nucleotide sequence of the chromosomal gene for human IL-4 and its expression.
We have isolated a chromosomal DNA segment of the human IL-4 gene based on homology with a human IL-4 cDNA sequence and determined its complete nucleotide sequence. The human IL-4 gene, which occurs as a single copy in the haploid genome, is mapped on chromosome 5. It is composed of four exons and three introns and is approximately 10 kilobase pairs in size. 5'-Flanking regions of human and mouse IL-4 genes share about 85% homology extending more than 500 base pairs upstream of a "TATA" like sequence. Several patches of sequences are found in the 5'-flanking region of the human IL-4 gene which are homologous to sequence in the 5'-flanking regions of the IL-2, IL-3, IL-5, and granulocyte-macrophage (GM)-CSF genes. The IL-4 gene is inducible after treatment of human T cell clone by phorbol-12-myristate-13-acetate (TPA) and calcium ionophore A23187. The 2.3-kb 5'-flanking region of the human IL-4 gene transiently transfected into Jurkat human T cell leukemia cells is activated efficiently in response to TPA and A23187 stimulation and, although less efficiently, by human T cell leukemia virus type I-encoded p40x or BPV-encoded E2 protein. Combination of TPA/A23187 and p40x or E2 protein further augmented the level of expression.
Amino Acid Sequence↗
Localization of human phosphoribosylpyrophosphate synthetase subunit I and II genes (PRPS1 and PRPS2) to different regions of the X chromosome and assignment of two PRPS1-related genes to autosomes.
Complementary DNA clones for phosphoribosylpyrophosphate synthetase subunits I and II (PRS I and PRS II) were used to determine the chromosomal localization of the corresponding human genes. Southern blot analysis of genomic DNAs isolated from human placenta and a panel of human-mouse somatic cell hybrids revealed that the rat PRS I cDNA probe detected at least five human specific DNA segments (23, 20, 14.5, 6.7, and 4.3 kb) in BamHI digests. The 23-, 14.5-, and 6.7-kb DNA segments were detected only if the hybrids contained human chromosome X or translocation chromosome 7p+ (7qter greater than 7p22::Xq21 greater than Xqter), indicating the location of these segments to Xq21-qter (PRPS1). The 20- and 4.3-kb DNA segments did not cosegregate with the other three segments, and spot blot hybridization analysis using flow-sorted human chromosomes indicated that these are the PRPS1-related genes (PRPS1L1 and PRPS1L2) and could be assigned to chromosomes 7 and 9, respectively. The human-specific PRS II cDNA probe revealed a BamHI DNA segment (17 kb), which segregated condordantly with the X chromosome but not with the PRPS1 gene. We surmise that the gene for PRS II (PRPS2) is located at a different region of the X chromosome, namely Xpter-a21.
Animals↗
Cadmium-induced synthesis of metallothioneins in human T and B cell purified by a fluorescence activated cell sorter.
Human peripheral blood lymphocytes were reacted with fluorescein-conjugated antibodies specific to T or B cell surface antigen and fractionated with a fluorescence activated cell sorter. The isolated T and B cells were examined for their capacity to synthesize metallothioneins (MTs). Analysis by gel electrophoresis indicated that both T and B cells were able to produce MTs in a Cd2+-inducible manner, suggesting that both cells types have a mechanism of protection against Cd2+ toxicity.
Adult↗
Subcellular localization of the EGF receptor maturation process.
The glycosylation and the processing of the epidermal growth factor (EGF) receptor are suggested to play a crucial role(s) in the activation of ligand binding activity. To examine whether the receptor acquires EGF binding activity in the endoplasmic reticulum (ER) or in the Golgi complex, we carried out parallel kinetic analysis of the EGF binding activity and the intracellular transport of the newly synthesized receptor by immunoprecipitation with the anti-EGF receptor antibody B4G7 using the EGF receptor hyperproducing cell line NA. The kinetic analysis revealed that a receptor capable of binding EGF appeared after 30 to 60 min labeling with [35S]methionine. Pulse-chase experiments also indicated that the receptor capable of binding EGF appeared after a 30-min pulse with a 30-min chase. Subcellular fractionation analysis indicated that the newly synthesized receptor was present in the Golgi complex after labeling with [35S]methionine for 30 min. After a 30-min chase, the Mr 170K receptor appeared in the Golgi complex and plasma membrane. Thus, these results together indicated that after a 30-min pulse incubation a fraction of the EGF receptors have been transported from the ER to the Golgi complex; however, the receptor is unable to bind EGF. Although the EGF receptor appeared on the cell surface after a 30-min pulse with a 30-min chase, only half of the receptors are capable of binding EGF. Therefore, the EGF receptor acquires ligand binding activity at a late stage of the maturation process, most likely in the Golgi complex.
Animals↗
Isolation of cDNAs encoding a substrate for protein kinase C: nucleotide sequence and chromosomal mapping of the gene for a human 80K protein.
An acidic phosphoprotein of Mr 80,000, the 80K protein, is a substrate for protein kinase C in fibroblasts and epidermal carcinoma cells. We purified the 80K protein from human squamous carcinoma Ca9-22 cells and fractionated it into two distinct molecular species, designated the 80K-L and 80K-H proteins. The amino acid sequences of the NH2-terminal region and cyanogen bromide-cleaved fragments of the 80K-H protein were determined and a corresponding oligonucleotide sequence was synthesized. Using this as a probe, two cDNA clones, lambda 80H-1 and lambda 80H-2, were selected from a lambda gt10 cDNA library from human A431 cells. The nucleotide sequence has an open reading frame of 1581 nucleotides encoding a protein of 527 amino acids. The deduced amino acid sequence revealed an extremely Glu-rich region. RNA blot analysis with the lambda 80H-1 cDNA clone detected two polyadenylated transcripts of 2.3 and 3.5 kb in Ca9-22 cells. Spot blot hybridization using flow-sorted human chromosomes provided evidence that the gene (G19P1) encoding 80K-H protein maps to human chromosome 19.
Amino Acid Sequence↗
The human ATP synthase beta subunit gene: sequence analysis, chromosome assignment, and differential expression.
In humans, the functional F0F1-ATP synthase beta subunit gene is located on chromosome 12 in the p13----qter region. Other partially homologous sequences have been detected on chromosomes 2 and 17. The bona fide beta subunit gene has 10 exons encoding a leader peptide of 49 amino acids and a mature protein of 480 amino acids. Thirteen Alu family DNA repeats are found upstream from the gene and in four introns. The gene has four "CCAAT" sequences upstream and in close proximity to the transcriptional initiation site. A 13-bp motif is found in the 5' nontranscribed region of both the beta subunit gene and an ADP/ATP translocator gene that is expressed in high levels in cardiac and skeletal muscle. Analysis of the beta subunit mRNA levels reveals marked differences among tissues. The highest levels are found in heart, lower levels in skeletal muscle, and the lowest levels in liver and kidney. These findings suggest that the tissue-specific levels of ATP synthase beta subunit mRNA may be generated through transcriptional control.
Amino Acid Sequence↗
[Fundamental study of 99mTc nanocolloids for inflammation imaging].
The biodistribution of nanometer-sized colloidal particles (nanocolloids, Solco-Basle Ltd.) was examined in six rabbits with turpentine-induced abscesses in the musculature of their hind legs. 99mTc labelled colloids with a mean particle size less than 100 nm were administered to each rabbit intravenously. The biodistribution of the tracer was studied with a gamma camera 1, 3, and 24 hours after injection. Prominent uptakes were demonstrated in the liver, spleen, bone marrow, kidneys, and urinary bladder. On the other hand, the abscess and surrounding inflammatory edema were depicted as unclearly margined faint hot spots. Subsequently, the rabbits were sacrificed and activity per gram tissues were counted with a well-type gamma counter. Although the results were almost in agreement with a gamma camera study, it was noticed the activity counts in the abscess were significantly higher than that of control muscles.
Abscess↗
[A case of systemic lupus erythematosus with neurological manifestations as initial symptoms].
According to the past reports, neuropsychiatric manifestations have been seen in 10-75% of patients with systemic lupus erythematosus and are second only to renal involvement as a cause of death. The clinical feature is multiple. And cerebrovascular diseases due to systemic lupus erythematosus are detected in 3-16% of the neuropsychiatric manifestations. Occlusion of the intracranial major arteries is less frequently found in other cerebrovascular diseases. And central nervous system involvement usually occurs at some intermediate or terminal stage of systemic lupus erythematosus, so is rarely regarded as one of the initial symptoms. We studied the case of a patient with systemic lupus erythematosus with occlusion of the right middle cerebral artery indicated by angitis and 'string of beads' appearance of the right internal carotid artery indicated by fibromuscular dysplasia. The patient was a 38 year old female and began to feel weakness in the left hand and developed mild-left hemiparesis due to infarction of right temporo-parieto-occipital lesion which was revealed by CT scan. Carotid angiograph showed irregularity at the right middle cerebral artery and 'string of beads' appearance of the right internal carotid artery. Gradually neurological manifestations improved, but a facial 'butterfly' rash, palmar erythema and polyarthritis were detected.(ABSTRACT TRUNCATED AT 250 WORDS)
Adult↗