Detection of a new weak A blood-group allele (Aw11).

BACKGROUND AND OBJECTIVES: Weak ABO variants may escape tests using unlicensed sera. MATERIALS AND METHODS: Prior to transfusion, ABO grouping was performed using an automated system and in-house diluted sera, and manual and bedside test techniques. Genotyping and sequencing were performed using standard methods. RESULTS: Initially, the red blood cells (RBC) of the first-time blood donor were typed as B, but pretransfusion testing carried out using the bedside test indicated the presence of an additional A phenotype. Serological re-examination confirmed the bedside test results, and the allele in question was identified, by genotyping, as a new weak A variant (Aw11). CONCLUSIONS: The use of CE-marked and licensed antisera is recommended to avoid ABO mistyping.

Identification and characterization of a novel HLA-DRB1 allele, DRB1*0830*.

Here, we report on the identification and characterization of a novel human leukocyte antigen (HLA) DRB1 allele, DRB1*0830. This allele was identified in a candidate for hematopoietic stem cell donation. While the DNA sequence of HLA-DRB1*0830 most closely matches to DRB1*080202, its amino acid sequence resembles DRB1*080202 and DRB1*0813. Due to a substitution at nucleotide position 286 HLA-typing using sequence-specific oligonucleotide hybridization or amplification using sequence-specific primers gave inconclusive results. Allele-specific DNA sequencing confirmed a 286 T-->A substitution resulting in Phe67Ile.

Human platelet antigen genotyping by using sequence-specific primers and the particle gel agglutination assay.

BACKGROUND AND OBJECTIVES: Polymerase chain reaction using sequence-specific primers (PCR-SSP) is currently the most widely used technique for human platelet antigen (HPA) genotyping. Here, we describe a novel particle gel-agglutination technique for simplified visualization of the amplified products. MATERIALS AND METHODS: Biotinylated primers were used to amplify HPA-1, -2, -3, -4, -5, -6, and -15, and the PCR products were incubated with streptavidin particles. Fluorescein isothiocyanate (FITC)-labelled primers [amplifying a fragment of the human growth hormone (HGH) gene] and anti-FITC-coated particles were used as internal controls. Agglutination of the particles in or on top of the gel indicated specific amplification. A total of 100 samples from blood donors was tested by using this new technique and a standard PCR-SSP protocol. RESULTS: The use of biotinylated sequence-specific primers resulted in PCR products that agglutinated streptavidin particles, and the FITC-labelled HGH primers led to agglutination of anti-FITC-coated particles. Negative reactions were clearly distinguishable from positive reactions. The results of the particle gel agglutination method were in concordance with those of the electrophoretic visualization in all cases tested. CONCLUSIONS: The new particle agglutination method is reliable and easy to use.

B*4440: a novel HLA-B allele identified by sequence-specific oligonucleotide hybridization and sequence-specific amplification.

A novel human leukocyte antigen B (HLA-B) allele, B*4440, is described. The allele was identified in an adult stem cell donor of Caucasian origin. HLA-B*4440 most closely matches to B*4403 differing by a substitution of three nucleotides at codon 44, 45, and 50. Thus, low-resolution HLA typing using sequence-specific oligonucleotide hybridization or amplification using sequence-specific primers gave inconclusive results. DNA sequencing confirmed a variation of codons 44 and 45 (AGG AAG-->AGA GAG) and codon 50 (CCA-->CCG), resulting in an amino acid substitution Lys-->Glu at codon 45.

Description of a novel HLA-B allele, B*5613, identified during HLA-typing using sequence-specific oligonucleotide hybridization and sequence-specific amplification.

Here, we report on the characterization of a novel human leukocyte antigen (HLA)-B allele, B*5613. The allele was identified in an adult male from North Africa who was suffering from sickle cell anemia. HLA-B*5613 most closely matches to B*5601 differing only by a substitution of three nucleotides of codon 180. Due to this substitution, low-resolution HLA-typing using sequence-specific oligonucleotide hybridization or amplification using sequence-specific primers gave inconclusive results. DNA sequencing confirmed a variation of codon 180 (CTG-->GAC) resulting in an amino acid substitution Leu156Asp.

Frequencies of alpha4 A3061G variants and identification of three new variants of the human integrin alpha4-subunit.

Human integrin alpha4beta1 (VLA-4) is thought to play a key role in immune responses. Only two variants of alpha4-subunit (alpha4-tex and alpha4-mas) have been described until now, and the information regarding these variants is scanty. In this study, we measured the frequency of both variants in healthy blood donors (n=252). Surprisingly, the frequency of alpha4-mas (3061G, 0.31) was lower than that of alpha4-tex (3061A, 0.69). In addition, we sequenced and analyzed the mRNA of the entire alpha4-subunit in eight unselected healthy blood donors. These studies revealed three new variants, including a C to A transversion at position 269 in the promoter region of exon 1 (accession no. AJ504733); a G to A transversion at position 2273 in exon 16 (accession no. AJ510246, AJ510247), and a T to C exchange at position 3311 in exon 26 (accession no. AJ510248, AJ510249). None of these variants led to amino acid (AA) substitutions. Interestingly, homozygosity of the new variant 269A was not found.

Donor-derived alloantibodies and passenger lymphocyte syndrome in two of four patients who received different organs from the same donor.

BACKGROUND: Reported here is the occurrence of RBC alloimmunization in two of four patients who received different organs from an immunized donor. STUDY DESIGN AND METHODS: The donor, a 58-year-old woman, was group O D+, K-, and Fy(a-). Initially, her serum contained only a K antibody. After blood transfusion, a second antibody (anti-Fy(a)) could also be identified. The liver was given to a group O D+, K-, Fy(a+) patient; the pancreas and one kidney to a group O D+, K-, Fy(a+) patient; the heart to a group A D+, K-, Fy(a-) patient; and the other kidney to a group B D+, K-, Fy(a+) patient. RBC grouping and antibody screening were performed by standard techniques. Lymphoid microchimerism in the peripheral blood of the recipients was analyzed by flow cytometry and nested PCR. RESULTS: None of the recipients had irregular RBC alloantibodies at the time of transplantation. After the transplant, anti-K became detectable in the serum of the liver recipient, and anti-Fy(a) could be eluted from the RBCs of the liver recipient and the pancreas-kidney recipient. The latter patient also developed mild hemolysis, and his Hb dropped to 8 g per dL on posttransplant Day 9. Donor-derived lymphocytes were detectable by flow cytometry in the peripheral blood of the liver recipient and the pancreas-kidney recipient until Days 8 and 63, respectively, whereas no lymphoid chimerism could be demonstrated in the heart recipient. PCR chimerism analyses were positive in all three recipients over the whole observation period of 97 postoperative days. CONCLUSION: The amount of cotransplanted lymphoid tissue may correlate with the extent of peripheral lymphoid microchimerism and the antibody-formation capacity in solid organ transplantation.

Percutaneous exposure to ethyl parathion in a feral Griffon vulture (Gyps fulvus).

A feral Griffon vulture (Gyps fulvus) was found with tremors, weakness, digit and wing flexion, and an inability to fly. A zero blood cholinesterase activity and a favorable response to treatment with pralidoxime hydrochloride indicated exposure to an anticholinergic pesticide. The bird died after 7 d, and traces of the organophosphate insecticide ethyl parathion were found in the liver and from a blue discolored skin area of the neck. Continuous exposure to ethyl parathion through dermal absorption was presumed the cause of death of the vulture.

A survey of hospital services for suicidal persons.

A person who threatens or attempts suicide, arriving for treatment at any 1 of 10 hospitals serving a suburban area in the Bay Area of California, will be met with a variety of responses by different kinds of personnel, depending on the hospital to which he comes. In half of the hospitals, he could be subsequently discharged without psychiatric consultation of any kind. Follow-through on any outpatient referral would usually be left up to him. Although a few hospitals attend to the problem of suicidal persons in many important respects, other hospitals pay little attention to these same aspects. Because of the important role of the hospital in the prevention of suicide, it is recommended that the hospital's function in dealing with suicidal patients be better defined, extended, and made more consistent.