HLA-DQ beta chain DNA restriction fragments can differentiate between healthy and narcoleptic individuals with HLA-DR2.
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Biomedical subjects
Publications and source records attributed to A Ando.
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Three homozygous cell lines with different cellular HLA-D specificities associated with HLA-DR2 including Dw2, Dw12, DB9 displayed structural variation in the products of the DQ locus. Nonglycosylated precursor polypeptides of DQ molecules immunoprecipitated by two-dimensional polyacrylamide gel electrophoresis (2-D gel) revealed distinct differences in the patterns of DQ alpha and DQ beta chains.
Restriction fragment-length polymorphisms (RFLP) were systematically analyzed by Southern hybridization with restriction endonuclease-digested genomic DNA from 28 HLA-homozygous B cell lines with Dw1-Dw19 specificity using the DR beta and DQ beta chain cDNAs as probes. These probes detected polymorphic fragments unique to each HLA-DR specificity. Furthermore, the DQ beta chain probes permitted us to distinguish between different Dw specificities with an identical DR type much more efficiently than with the DR beta chain probe. Distribution analysis of restriction fragments hybridizing to DR beta in relation to the DR and DQ specificities showed several sets of them forming ten clusters, some of which correlate with DRw53, DQw1, and DR alleles. This DNA typing technique allows the direct definition of HLA types at the gene level and provides a powerful tool for isolating genes controlling HLA-associated diseases.
From a human cDNA library constructed from consanguineous HLA-homozygous cell line AKIBA (HLA-A24, Bw52, DR2, Dw12, Cp63 Cp63: a new DP type), cDNA clones encoding the heavy and light chains of an HLA-DP Cp63 alloantigen were isolated and analyzed by restriction enzyme mapping and nucleotide sequence determination. Allelic comparisons of DP alpha and DP beta cDNA sequences showed that the amino acid sequence of the DP alpha chain was less polymorphic than that of the DP beta chain. In the DP beta chain, the polymorphic region was restricted to the beta 1 domain. We also isolated and characterized 15 genomic phage clones spanning a 74 kilobase (kb) pair of the DP region which were found to contain one DP alpha gene, one DP-like alpha gene, one DP beta gene, and one DP-like beta gene. Genomic blot analyses with different HLA-DP type cell lines using DP alpha cDNA as a probe revealed EcoRI fragment length polymorphism around the DP alpha gene.
Tumor uptake rates, concentrations in the mitochondrial fraction (containing lysosome) of liver and tumors, avid accumulations in connective tissue (especially inflammatory tissue) and binding substances in these tissues of 95Zr and 181Hf were essentially similar to those for 67Ga, 111In, 169Yb and 167Tm. However, the main binding substance of the above elements in group IV in tumor and liver was acid mucopolysaccharide whose molecular weight exceeded 40,000, although the above elements in group III were bound mainly to the acid mucopolysaccharide with a molecular weight of about 10,000.
A new method has been developed for imaging the thyroid gland by the use of 201Tl radioactive implant induced X-ray emission (RIXE). The thyroid phantom was made from two cylindrical tubes embedded in plastic neck phantom. Each tube consisted of stable iodine and 201Tl radioactive source in 10 ml water solution. Some photopeaks in the energy range from 20 to 200 ke V were scanned along a horizontal axis by the collimated Ge detector with high energy resolution. The distribution on the horizontal axis was obtained for the generated I K alpha-ray and some primary 201Tl radiations. The 201Tl RIXE scanning can provide information about the distribution of iodine with 201Tl as well as 201Tl distribution in the thyroid gland. The effectiveness of the 201Tl RIXE technique as a thyroid scanning method is discussed.
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Accumulation of oxalate, resulting in high plasma levels, is a common finding in end-stage renal disease. We investigated plasma concentration and peritoneal clearance of oxalate in 14 patients on continuous ambulatory peritoneal dialysis. The plasma oxalate levels in these patients (30.2 +/- 11.2 mumol/l) were as high as those in hemodialysis patients before dialysis (31.9 +/- 11.1 mumol/l). There was a significant correlation between plasma oxalate and urea nitrogen appearance (UNA). Dietary protein seems to be an important oxalate source in these patients, because the UNA reflects protein intake in stable patients. The mean peritoneal oxalate clearance was 6.64 +/- 1.56 l/day, close to the creatinine clearance. These results suggest that the plasma oxalate levels in CAPD patients may be sufficiently high to induce calcium oxalate deposition, and that methods of increasing oxalate removal and reducing oxalate burden are necessary for CAPD patients.
Using normal rats, retention values and subcellular distribution of 67Ga in each organ were investigated. At 10 min after administration of 67Ga-citrate the retention value of 67Ga in blood was 6.77% dose/g, and this value decreased with time. The values for skeletal muscle, lung, pancreas, adrenal, heart muscle, brain, small intestine, large intestine and spinal cord were the highest at 10 min after administration, and they decreased with time. Conversely, this value in bone increased until 10 days after injection. But in the liver, kidney, and stomach, these values increased with time after administration and were highest 24 h or 48 h after injection. After that, they decreased with time. The value in spleen reached a plateau 48 h after administration, and hardly varied for 10 days. From the results of subcellular fractionation, it was deduced that lysosome plays quite an important role in the concentration of 67Ga in small intestine, stomach, lung, kidney and pancreas; a lesser role in its concentration in heart muscle, and hardly any role in the 67Ga accumulation in skeletal muscle. In spleen, the contents in nuclear, mitochondrial, microsomal, and supernatant fractions all contributed to the accumulation of 67Ga.
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After administration 67Ga concentrates with time in lysosomes from the cytoplasm of liver cells. The lysosomal role in the accumulation of 67Ga in the liver cell is weakened upon transformation of the liver cell into a malignant tumor cell. In malignant tumors (except for hepatoma) the lysosome does not play a major role in the tumor concentration of 67Ga. 67Ga is bound to acid mucopolysaccharides (keratan polysulfate, etc.) in both tumor and liver. In liver cells, large amounts of 67Ga are transported into lysosomes with these acid mucopolysaccharides, and in hepatoma cells, quite large amounts of 67Ga are transported into lysosomes with these acid mucopolysaccharides. In malignant tumor cells (except for hepatoma) the effect is much smaller, the acid mucopolysaccharides transporting very little 67Ga into lysosome. The 67Ga is concentrated in viable tumor tissue within malignant tissue but hardly at all in necrotic tumor tissue, and concentrates avidly in inflammatory infiltration around tumor cells. Plenty of 67Ga is found in liver but very little in connective tissue associated with the liver.
Normal male rats were injected with either gallium citrate Ga 67 or sodium sulfate S 35. After 24 h, the stomach, small intestine, pancreas, and muscle were excised and homogenized. After the removal of the nuclear fraction, each of these homogenates was digested with protease. After digestion, the supernatants of the reaction mixtures were applied to a Sephadex-G-100 column. The radioactivity was eluted with buffer solution. The resultant eluates were analyzed for radioactivity and the levels of proteins, uronic acids, and sialic acids. In all four organs, sizable amounts of 67Ga were bound to sulfated acid mucopolysaccharides with molecular masses of about 10,000 daltons and to sulfated acid mucopolysaccharides, a species whose molecular masses exceed 40,000 daltons. In the stomach, large amounts of 67Ga were bound to sulfated acid mucopolysaccharides with molecular masses of about 10,000 daltons. From these results, it is obvious that the main 67Ga-binding substances in these four organs are sulfated acid mucopolysaccharides, and that these acid mucopolysaccharides play the most important role in the concentration of 67Ga in these organs.
The bipositive ions and anions, with few exceptions, indicated a low tumor uptake rate. On the other hand, compounds of Hg, Au and Bi, which have a strong binding power to protein, showed a high tumor uptake rate. As Hg2+, Au+ and Bi3+ are soft acids according to the classification of Lewis acids, it was thought that these ions would bind strongly to soft bases (R-SH, R-S-) present in tumor tissue. For many hard acids such as 85Sr2+, 67Ga3+, 181Hf4+, and 95Nb5+, tumor uptake rates are shown as a function of ionic potentials (valency/ionic radii) of the metal ions. Considering the present data and previously reported results, it was presumed that hard acids of trivalence, quadrivalence and pentavalence would replace calcium in the calcium salts of hard bases (calcium salts of acid mucopolysaccharides, etc.). Ionic potentials of alkaline metals and Tl were small, but the tumor-uptake rate of these elements indicated various values. As Ge and Sb are bound by covalent bonds to chloride, GeCl4 and SbCl3 behaved differently from many metallic compounds in tumor tissue.
It was determined from measuring neutral saccharide in the structure that the principal 67Ga-binding acid mucopolysaccharide in liver was keratan sulfate and/or keratan polysulfate. On the other hand, it was clarified from the results of mucopolysaccharase treatment that the main 67Ga-binding acid mucopolysaccharide in liver was not either one of keratan sulfate, heparan sulfate, heparin, chondroitin sulfate A, B and C. Based on the present results, it was deduced that the main 67Ga-binding acid mucopolysaccharide in liver was keratan polysulfate.
The gene expression of myelin basic proteins (MBPs) in shiverer mutant mice was investigated by the Northern and Southern hybridization techniques. In the control mice RNA molecules from the brains which were about 2,300 nucleotides in length were hybridized to cDNA of 1.8 kb encoding for a mouse MBP, but RNA from the brains of 3-week-old shiverer mutant mice contained no detectable amount of MBP transcripts hybridizing to this probe. Moreover the shiverer mutant mice lost several restriction fragments that hybridized to the same probe in the control mice when each of the five restriction enzymes, i.e., HindIII, PstI, PvuII, AccI, and StuI, was used. These data suggest that the shiverer mutation may correspond to the deletion of a large portion of MBP exon(s) in the gene, and this deletion causes inefficient transcription leading to the depletion of MBPs in the myelin and the dysmyelination observed in these mice.
In the previous paper, we reported that 67Ga was accumulated in abscess and uptake rate of 67Ga in abscess increased with time after the injection of 67Ga-citrate. The present study was undertaken to elucidate the influence of blood flow on the accumulation of 67Ga in abscess. Five days after subcutaneous injection of 0.2 ml of turpentine to the rats, 131I-human serum albumin (HSA) was injected intravenously to the rats. At an appropriate time after the injection (10 min to 6 days), uptake rates of 131I-HSA in abscess and normal tissues were measured. Similarly, 51Cr-red blood cells (RBC) were injected intravenously to the above rats and the uptake rates of 51Cr-RBC were also measured. One, three, and 24 hours after injection of 131I-HSA, the uptake rates of 131I-HSA in abscess were 1.32 %dose/g, 1.84 %dose/g, and 0.82 %dose/g, respectively. However, the uptake rates of 51Cr-RBC in abscess was very small, and the value was 0.14 %dose/g at 24 hours after the injection. In the case of abscess, blood in the tissue fluid was very little, but the permeability of 131I-HSA from the blood vessel in the tissue was much larger than that of normal tissues. From these facts, it was deduced that the accelerated permeability caused the abscess accumulation of 67Ga.