Current human leukocyte antigen (HLA) DNA typing methods such as the sequence-based typing (SBT) and sequence-specific oligonucleotide (SSO) methods generally yield ambiguous typing results because of oligonucleotide probe design limitations or phase ambiguity for HLA allele assignment. Here we describe the development and application of the super high-resolution single-molecule sequence-based typing (SS-SBT) of HLA loci at the 8-digit level using next generation sequencing (NGS). NGS which can determine an HLA allele sequence derived from a single DNA molecule is expected to solve the phase ambiguity problem. Eight classical HLA loci-specific polymerase chain reaction (PCR) primers were designed to amplify the entire gene sequences from the enhancer-promoter region to the 3' untranslated region. Phase ambiguities of HLA-A, -B, -C, -DRB1 and -DQB1 were completely resolved and unequivocally assigned without ambiguity to single HLA alleles. Therefore, the SS-SBT method described here is a superior and effective HLA DNA typing method to efficiently detect new HLA alleles and null alleles without ambiguity.
The pathogenesis of histamine-induced coronary spasm was examined angiographically and morphometrically in Gbttingen miniature pigs. In five of 36 consecutive pigs that were 4 to 5 months of age, coronary spasm was provoked by the intracoronary administration of histamine, and the left coronary arteries were examined histologically without endothelial denudation (group 1). Endothelial balloon denudation of the major branch of the left coronary artery was performed in 31 of 36 pigs and five died during the procedure. The remaining 26 pigs were randomly allotted to one of two groups, one fed a cholesterol-supplemented (group 2, n = 13) and one fed a regular low-cholesterol diet (group 3, n = 13). After 3 months, serum cholesterol increased significantly from 57 +-6 to 222 + 27 mg/dl (p < .01) in group 2, but remained unchanged (48 + 5 to 55 6 mg/dl) in group 3. Percent narrowing of the coronary diameter induced by 10 gg/kg ic histamine after administration of the H2 blocker cimetidine (60 mg/kg iv) was 39 + 3% and 24 ± 2% (p < .05 between groups 2 and 3) at the nondenuded site and 78 ± 3% and 74 + 4% at the denuded site in groups 2 and 3, respectively (p < .01 between nondenuded and denuded sites). Histamine-induced percent narrowing of the coronary diameter after cimetidine in group 1, 2, and 3 pigs correlated well with the degree of intimal thickness on an exponential curve (r = .92, p < .001). Since percent narrowing at the intact site was 27% (n = 19) in all three groups, predicted histamine-induced percent narrowing at the spastic site, applying the geometric theory, was 33 ± 3%. Accordingly, enhanced constriction of the coronary artery with intimal thickening in response to histamine can largely be explained by the acquired hyperresponsiveness of the vascular wall to autacoids. This phenomena, not related to the level of serum cholesterol, may be uniquely linked to the basic pathology of evolution of atherosclerosis. Circulation 74, No. 4, 826-837, 1986. RECENTLY, enhanced responsiveness of the medium-sized arteries to vasoactive substances has been noted not only clinically in patients with variant angina,'-' but also experimentally in vessels exposed to cholesterol6 or hypercholesterolemia,' 8 in atheroscle-
The relationship between average grain size on the surface of SnO 2 transparent conductive film and conversion efficiency of the a-Si:H solar cell was investigated. a-Si:H solar cells were fabricated on SnO2/glass substrates with various grain sizes.The cell structure was glass/p(SiC)-i-n/Al and the effective cell area was 4 x 10 -2cm2. The reflectivity from the glass substrate was reduced to about 7 percent with increasing the grain size from 0.1 to 0.8fim. and the short-circuit current was inceased from 12 to 14mA/cm2. A 7.9 percent of conversion efficiency was achieved using milky S n 0 2 film of 0 . 4 -~m average grain size at AM-lOOrnWicm2.T
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