BACKGROUND
Anti‐KANNO, a broadly reactive RBC alloantibody, is found among some Japanese pregnant women, but the genetic basis of the corresponding antigen remains unclear.
STUDY DESIGN AND METHODS
...We integrated a statistical approach to identify the coding gene for KANNO antigen by conducting a genome‐wide association study (GWAS) on four KANNO‐negative individuals and 415 healthy Japanese. We also applied whole‐exome sequencing to them and performed a replication study to confirm the identified genome variation using independent 14 KANNO‐negative individuals. A monoclonal antibody‐specific immobilization of erythrocyte antigens (MAIEA) assay was used to locate KANNO antigen on RBC‐specific membrane protein. In vivo and in vitro binding assays of anti‐KANNO were further applied to the cells expressing a candidate protein.
RESULTS
The GWAS revealed a genome‐wide significant association of chromosome 20p13 locus (p = 2.76E‐08; odds ratio > 1000 95% confidence interval = 48–23,674). The identified single‐nucleotide polymorphism located in an intronic region of the prion protein (PRNP) gene. Whole‐exome sequencing revealed a missense variant in the PRNP gene (rs1800014, E219K), which is in linkage disequilibrium with the single‐nucleotide polymorphism identified in the GWAS. All 18 KANNO‐negative individuals possessed the homozygous genotype of the missense variant. The MAIEA assay using anti‐KANNO and mouse antihuman prion protein showed a clear difference between KANNO‐positive and KANNO‐negative RBCs. Anti‐KANNO showed direct binding to CHO‐K1 cells expressing wild‐type PRNP but not to those expressing E219K PRNP.
CONCLUSION
We first identified the coding gene of the high‐frequency antigen KANNO located in PRNP and the missense variation (E219K) that affects the seropositivity of the KANNO antigen, which were confirmed by PRNP overexpressed cells.
Glycophorin hybrids such as GP.Mur are common in Southeast Asians. In Taiwan, clinically significant alloantibodies to the GP.Mur phenotype are the most important issue in blood banks. A large-scale ...screening of glycophorin hybrids in the Taiwanese population is urgently needed to ensure transfusion safety. Four clones of human hybridomas that secrete anti-Mia, anti-MUT, and anti-Mur were established by fusing human B-lymphocytes and myeloma cells (JMS-3). The specificity of each monoclonal antibody (MoAb) was characterized. Three MoAbs were applied on an Automated Pretransfusion Blood Testing Analyzer (PK7300/PK7400) for donor screening. Genotyping was performed to determine the detailed subgrouping of glycophorin hybrids. Four MoAbs are IgM antibodies. Anti-Mia (377T) binds to 46DXHKRDTYA54, 48HKRDTYAAHT57 peptides, and anti-Mia (367T) binds to 43QTNDXHKRD51 peptides (X indicates T, M, or K). Anti-Mur is reactive with 49KRDTYPAHTA58 peptides. Anti-MUT is reactive with 47KHKRDTYA54. A total of 78,327 donors were screened using three MoAbs, and 3690 (4.71%) were GP.Mur, 20 (0.025%) were GP.Hut, and 18 (0.022%) were GP.Vw. When the Mia antigen was introduced as routine screening, the frequency of Mi(a+) among blood donors in Taiwan was 4.66% (67,348/1,444,541). Mia antigen was implemented as a routine blood testing, and the results were labeled on all red blood cell (RBC) units.
BACKGROUND
MNS is one of the highly polymorphic blood groups comprising many antigens generated by genomic recombination among the GYPA, GYPB, and GYPE genes as well as by single‐nucleotide changes. ...We report a patient with red blood cell (RBC) antibody against an unknown low‐frequency antigen, tentatively named SUMI, and investigated its carrier molecule and causal gene.
STUDY DESIGN AND METHODS
Standard serologic tests, including enzyme tests, were performed. Monoclonal anti‐SUMI–producing cells (HIRO‐305) were established by transformation and hybridization methods using lymphocytes from a donor having anti‐SUMI. SUMI+ RBCs were examined by immunocomplex capture fluorescence analysis (ICFA) using HIRO‐305 and murine monoclonal antibodies against RBC membrane proteins carrying blood group antigens. Genomic DNA was extracted from whole blood, and the GYPA gene was analyzed by polymerase chain reactions and Sanger sequencing.
RESULTS
Serologic screening revealed that 23 of the 541,522 individuals (0.0042%) were SUMI+, whereas 1351 of the 10,392 individuals (13.0%) had alloanti‐SUMI. SUMI antigen was sensitive to ficin, trypsin, pronase, and neuraminidase, but resistant to α‐chymotrypsin and sulfydryl‐reducing agents. ICFA revealed that the SUMI antigen was carried on glycophorin A (GPA). According to Sanger sequencing and cloning, the SUMI+ individuals had a GYPA*M allele with c.91A>C (p.Thr31Pro), which may abolish the O‐glycan attachment site.
CONCLUSIONS
The new low‐frequency antigen SUMI is carried on GPA encoded by the GYPA*M allele with c.91A>C (p.Thr31Pro). Neuraminidase sensitivity suggests that glycophorin around Pro31 are involved in the SUMI determinant.
BACKGROUND
MNS is one of the highly polymorphic blood groups comprising many antigens generated by genomic recombination among the
GYPA
,
GYPB,
and
GYPE
genes as well as by single‐nucleotide changes. ...We report a patient with red blood cell (RBC) antibody against an unknown low‐frequency antigen, tentatively named SUMI, and investigated its carrier molecule and causal gene.
STUDY DESIGN AND METHODS
Standard serologic tests, including enzyme tests, were performed. Monoclonal anti‐SUMI–producing cells (HIRO‐305) were established by transformation and hybridization methods using lymphocytes from a donor having anti‐SUMI. SUMI+ RBCs were examined by immunocomplex capture fluorescence analysis (ICFA) using HIRO‐305 and murine monoclonal antibodies against RBC membrane proteins carrying blood group antigens. Genomic DNA was extracted from whole blood, and the
GYPA
gene was analyzed by polymerase chain reactions and Sanger sequencing.
RESULTS
Serologic screening revealed that 23 of the 541,522 individuals (0.0042%) were SUMI+, whereas 1351 of the 10,392 individuals (13.0%) had alloanti‐SUMI. SUMI antigen was sensitive to ficin, trypsin, pronase, and neuraminidase, but resistant to α‐chymotrypsin and sulfydryl‐reducing agents. ICFA revealed that the SUMI antigen was carried on glycophorin A (GPA). According to Sanger sequencing and cloning, the SUMI+ individuals had a
GYPA*M
allele with c.91A>C (p.Thr31Pro), which may abolish the O‐glycan attachment site.
CONCLUSIONS
The new low‐frequency antigen SUMI is carried on GPA encoded by the
GYPA*M
allele with c.91A>C (p.Thr31Pro). Neuraminidase sensitivity suggests that glycophorin around Pro31 are involved in the SUMI determinant.
Background and Objectives
Antigens of the MNS blood group system are expressed on the red blood cell (RBC) membrane on glycophorin A (GPA) and glycophorin B (GPB) or on hybrid molecules of GPA and ...GPB. This study investigated the distribution of glycophorin variants and alloantibodies against Hil and MINY among Japanese individuals.
Methods
Mi(a+) or Hil+ RBCs were screened using an automated blood grouping machine (PK7300) with monoclonal anti‐Mia or polyclonal anti‐Hil. Glycophorin variants were defined by serology with monoclonal antibodies against Mia, Vw, MUT and Mur, and polyclonal antibodies against Hil, MINY and Hop + Nob (KIPP). The glycophorin variants were further confirmed by immunoblotting and Sanger sequencing. Alloanti‐Hil and alloanti‐MINY in the plasma were screened using GP.Hil RBCs in an antiglobulin test. The specificity of anti‐Hil or anti‐MINY was assessed using GP.Hil (Hil+MINY+) and GP.JL (Hil‒MINY+) RBCs.
Results
The GP.HF, GP.Mur, GP.Hut, GP.Vw, GP.Kip and GP.Bun frequencies in 1 005 594 individuals were 0·0357%, 0·0256%, 0·0181%, 0·0017%, 0·0009% and 0·0007%, respectively. GP.Hil was found in as four of the 13 546 individuals (0·0295%). Of 137 370 donors, 10 had anti‐Hil (0·0073%) and three had anti‐MINY (0·0022%).
Conclusions
Glycophorin variants were relatively rare in Japanese individuals, with the major variants being GP.HF (0·0357%), GP.Hil (0·0295%) and GP.Mur (0·0256%). Only one example of anti‐MINY was previously reported, but we found three more in this study.
Glycophorin hybrids such as GP.Mur are common in Southeast Asians. In Taiwan, clinically significant alloantibodies to the GP.Mur phenotype are the most important issue in blood banks. A large-scale ...screening of glycophorin hybrids in the Taiwanese population is urgently needed to ensure transfusion safety. Four clones of human hybridomas that secrete anti-Mi
, anti-MUT, and anti-Mur were established by fusing human B-lymphocytes and myeloma cells (JMS-3). The specificity of each monoclonal antibody (MoAb) was characterized. Three MoAbs were applied on an Automated Pretransfusion Blood Testing Analyzer (PK7300/PK7400) for donor screening. Genotyping was performed to determine the detailed subgrouping of glycophorin hybrids. Four MoAbs are IgM antibodies. Anti-Mi
(377T) binds to
DXHKRDTYA
,
HKRDTYAAHT
peptides, and anti-Mi
(367T) binds to
QTNDXHKRD
peptides (X indicates T, M, or K). Anti-Mur is reactive with
KRDTYPAHTA
peptides. Anti-MUT is reactive with
KHKRDTYA
. A total of 78,327 donors were screened using three MoAbs, and 3690 (4.71%) were GP.Mur, 20 (0.025%) were GP.Hut, and 18 (0.022%) were GP.Vw. When the Mi
antigen was introduced as routine screening, the frequency of Mi(a+) among blood donors in Taiwan was 4.66% (67,348/1,444,541). Mi
antigen was implemented as a routine blood testing, and the results were labeled on all red blood cell (RBC) units.
Hydrophilic poly(2-(methacryloyloxy)ethyl phosphorylcholine) (PMPC) shows biocompatibility because the pendant phosphorylcholine group has the same chemical structure as the hydrophilic part of ...phospholipids that form cell membranes. Hollow particles can be used in various fields, such as a carrier in drug delivery systems because they can encapsulate hydrophilic drugs. In this study, vinyl group-decorated silica particles with a radius of 150 nm were covered with cross-linked PMPC based on the graft-through method. The radius of PMPC-coated silica particles increased compared to that of the original silica particles. The PMPC-coated silica particles were immersed in a hydrogen fluoride aqueous solution to remove template silica particles to prepare the hollow particles. The PMPC hollow particles were characterized by dynamic light scattering, infrared spectroscopy, thermogravimetric analysis, and transmission electron microscopy observations. The thickness of the hollow particle shell can be controlled by the polymerization solvent quality. When a poor solvent for PMPC was used for the polymerization, PMPC hollow particles with thick shells can be obtained. The PMPC hollow particles can encapsulate hydrophilic guest molecules by immersing the hollow particles in a high-concentration guest molecule solution. The biocompatible PMPC hollow particles can be used in a drug carrier.
Urinary exosomes, small extracellular vesicles present in urine, are secreted from all types of renal epithelial cells. Aquaporin-2 (AQP2), a vasopressin-regulated water channel protein, is known to ...be selectively excreted into the urine through exosomes (UE-AQP2), and its renal expression is decreased in nephrotic syndrome. However, it is still unclear whether excretion of UE-AQP2 is altered in nephrotic syndrome. In this study, we examined the excretion of UE-AQP2 in an experimental rat model of nephrotic syndrome induced by the administration of puromycin aminonucleoside (PAN). Rats were assigned to two groups: a control group administered saline and a PAN group given a single intraperitoneal injection of PAN (125 mg/kg) at day 0. The experiment was continued for 8 days, and samples of urine, blood, and tissue were collected on days 2, 5, and 8. The blood and urine parameters revealed that PAN induced nephrotic syndrome on days 5 and 8, and decreases in the excretion of UE-AQP2 were detected on days 2 through 8 in the PAN group. Immunohistochemistry showed that the renal expression of AQP2 was decreased on days 5 and 8. The release of exosomal marker proteins into the urine through UEs was decreased on day 5 and increased on day 8. These data suggest that UE-AQP2 is decreased in PAN-induced nephrotic syndrome and that this reflects its renal expression in the marked proteinuria phase after PAN treatment.
Energy metabolism and substrate oxidation during sleep correlate with sleep stage, suggesting that energy metabolism affects sleep architecture or vice versa. The aim of the present study was to ...examine whether changes in energy metabolism during sleep, induced by a high-carbohydrate or high-fat meal for dinner, affect sleep architecture. Ten healthy males participated in this study, sleeping 3 nonconsecutive nights in a whole-room calorimeter. The first night was scheduled as an adaptation to the experimental environment. The other 2 nights were experimental calorimetry in a balanced cross-over design with intrasubject comparisons. In each session, subjects comsumed a high carbohydrate (HCD: PFC=10 : 10 : 80) or high fat (HFD: PFC=10 : 78 : 12) meal at 2000 h and slept with a polysomnographic recording in a metabolic chamber for indirect calorimetry (0000 h to 0800 h). Slow wave sleep was decreased during the first sleep cycle and not changed during the second or third sleep cycle under HCD conditions compared with those of HFD. Energy expenditure was not affected by dietary condition but substrate oxidation reflected differences in dietary composition of the dinner during the first and second sleep cycle. The present study suggested the possibility that substrate availability during sleep affects substrate oxidation during sleep, and affects sleep architecture during the first sleep cycle.
Morphological and functional comparison of convergently-evolved traits in marsupials and eutherians is an important aspect of studying adaptive divergence in mammals. However, the anatomy of ...marsupials has been particularly difficult to evaluate for multiple reasons. First, previous studies on marsupial anatomy are often uniformly old and non-exhaustive. Second, muscle identification was historically based on muscle attachment sites, but attachment sites have since been declared insufficient for muscle identification due to extensive interspecific variation. For example, different names have been used for muscles that are now thought to be equivalent among several different species, which causes confusion. Therefore, descriptions of marsupial muscles have been inconsistent among previous studies, and their anatomical knowledge itself needs updating. In this study, the koala was selected as the representative marsupial, in part because koala locomotion may comprise primate (eutherian)-like and marsupial-like mechanics, making it an interesting phylogenetic group for studying adaptive divergence in mammals. Gross dissection of the lower limb muscles (the gluteal and the posterior thigh regions) was performed to permit precise muscle identification. We first resolved discrepancies among previous studies by identifying muscles according to their innervation; this recent, more reliable technique is based on the ontogenetic origin of the muscle, and it allows for comparison with other taxa (i.e., eutherians). We compared our findings with those of other marsupials and arboreal primates and identified traits common to both arboreal primates and marsupials as well as muscle morphological features unique to koalas.