International Society of Blood Transfusion Working Party on red cell immunogenetics and blood group terminology: Cancun report (2012).

The International Society of Blood Transfusion Working Party on red cell immunogenetics and blood group terminology convened during the International congress in Cancun, July 2012. This report details the newly identified antigens in existing blood group systems and presents three new blood group systems.

Diagnostic accuracy of spot and timed measurements of urinary albumin concentration to determine microalbuminuria in sickle cell disease.

OBJECTIVE: Whereas measurement of albumin:creatinine ratio (ACR) in spot urine samples is indicated for determining microalbuminuria, its performance or that of urinary albumin excretion rate (UAER) in predicting microalbuminuria in sickle cell disease (SCD) is unclear. We therefore tested the diagnostic performance of these measures in spot and timed urine samples in predicting a UAER in 24-hour samples. METHODS: Thirty participants with SCD had spot, two-hour and four-hour, followed by 24-hour urine collections for ACR, urinary albumin concentration (UAC) and UAER determinations. Receiver operating characteristic (ROC) curve analyses were performed. RESULTS: The areas under the ROC curves for microalbuminuria were 0.99 (CI: 0.97, 1.00) for ACR and 0.97 (CI: 0.92, 1.00) for UAC in spot urine samples. For ACR, at the cut-point of 4.13 mg/mmol, there was 100% sensitivity and 82.6% specificity, allowing an 86.2% correct classification. At the cut-point of UAC = 20.9 mg/L, there was 100% sensitivity and 73.9% specificity, allowing a 79.3% correct classification. Corresponding areas for microalbuminuria in two-hour timed samples were 0.99 (CI: 0.95, 1.00) for ACR and 0.96 (CI: 0.89, 1.00) for UAER.For ACR, the cut-point was 4.64 mg/mmol with 83.3% sensitivity and 91.3% specificity, allowing an 89.7% correct classification. Similarly for UAER, at the cut-point of 21.8 µg/min, there was 83.3% sensitivity and 91.3% specificity, allowing 89.7% correct classification. CONCLUSIONS: The diagnostic performance of ACR and UAC in a spot as well as ACR and UAER in two-hour timed urine samples in patients with SCD is excellent. Healthcare professionals can confidently utilize these measures in this patient population.

Molecular basis of the rare gene complex, DIVa(C)-, which encodes four low-prevalence antigens in the Rh blood group system.

BACKGROUND: Over 40 years ago, an unusual Rh phenotype denoted DIVa(C)- was identified in a case of fatal haemolytic disease of the newborn in the third child of Madame Nou. Her RBCs expressed a partial D, weak C and four low-prevalence Rh antigens: Go(a) (RH30), Rh33 (RH33), Riv (RH45) and FPTT (RH50). The purpose of this study was to determine the molecular basis associated with this rare DIVa(C)- complex. MATERIAL AND METHODS: Blood samples were from three donors previously identified as carrying the DIVa(C)- haplotype. Molecular analyses were performed by standard methods. RESULTS: The three donors were heterozygous for RHD and RHD*DIVa.2, and all carried a compound hybrid allele at the RHCE locus. This hybrid RHCE allele contained exons 2 and 3 from RHD*DIVa.2 and exon 5 from RHD [RHCE*CE-DIVa.2(2-3)-CE-D(5)-CE] and is in cis to RHD*DIVa.2. The RHCE allele on the in trans chromosome differs between the donors and is RHCE*cE in donor 1, RHCE*ce (254C, 733G) in donor 2 and RHCE*ce in donor 3. CONCLUSIONS: The RHD*DIVa.2 encodes the Go(a) antigen, whereas the compound hybrid allele most likely encodes Rh33, Riv and FPTT. The weakly expressed C antigen on RBCs with the DIVa(C)- phenotype could be encoded by exons 2 and 3 from RHD*DIVa.2 in the compound hybrid. This is the first report of RHD*DIVa.2 being involved in a hybrid gene at the RHCE locus. As only one example of anti-Riv has been described, our molecular analysis and findings provide a tool by which to predict Riv expression.

The molecular basis of the LU:7 and LU:-7 phenotypes.

The Lutheran blood group system currently consists of 20 antigens that have been assigned ISBT numbers. Of these, all but LU7 have been associated with one or more nucleotide changes in LU. The purpose of this study was to determine the molecular basis associated with the LU:-7 phenotype. We obtained a stored sample from one proband with this phenotype and sequenced LU. Using genomic DNA, exons 1 through 15, and their flanking intronic regions, of LU were amplified by polymerase-chain reaction, and the products were sequenced. A homozygous novel missense nucleotide change of 1274A>C in exon 10 of LU was observed. This change is predicted to encode Ala at position 425 in place of Glu of the consensus Lu glycoprotein. Based on these results, and an absence of a record of this change in the Single Nucleotide Polymorphism database, Glu425 in the Lu glycoprotein is required for expression of Lu7, and Ala425 is associated with the LU:-7 phenotype. This completes the molecular basis associated with all antigens known to be in the Lutheran blood group system.

Molecular background of RH in Bastiaan, the RH:-31,-34 index case, and two novel RHD alleles.

Anti-hr(B) (-RH31) and anti-Hr(B) (-RH34) were found nearly 40 years ago in the serum of a South African woman. The anti-hr(B) was revealed after adsorption with DcE/DcE red blod cells (RCBs). Numerous anti-hr(B), in the absence of anti-Hr(B), have since been identified. We obtained a sample of blood from this index case (Bastiaan) and report the molecular basis of her D+C-E-c+e+/-, V-VS+Hr+hr(S)+hr(B)-Hr(B)- phenotype as well as results of testing her RBCs using currently available regents. We tested a cohort of African Americans to estimate the frequency of the RHCE*ce 48C,733G,1006T allele, and in addition found two novel RHD alleles. Hemagglutination tests and DNA analyses were performed by standard methods. Analyses revealed homozygosity for RHCE*ce 48C,733G,1006T in Bastiaan. RBCs from Bastiaan were strongly agglutinated by three commercial anti-e reagents. Testing RBCs from people homozygous for RHCE*ce 48C,733G,1006T showed that anti-e MS16, MS17, and MS63 were weakly reactive or non reactive, MS21 was strongly reactive, and HIRO38, HIRO41, and HIRO43 were non reactive. Results show that Bastiaan has RHD*DIIIa150C and RHD*DIIIa-CE(4-7)-D. Tests on 605 samples from random African Americans revealed a frequency of 0.036 for RHCE*ce 48C,733G,1006T and revealed two novel alleles: RHD*186T and RHD*DIIIa150C. The Bastiaan phenotype is encoded by RHD*DIIIa150C-RHCE*ce 48C,733G,1006T and RHD*DIIIa-CE(4-7)-D-RHCE*ce 48C,733G,1006T ; thus, this genotype is the gold standard for the hr(B)-Hr(B)-phenotype. The r'(s) complex encodes VS, which explains why most hr(B)-RCBs are VS+.

The production, serologic evaluation, and epitope mapping of ten murine monoclonal Dombrock antibodies.

The Dombrock (Do) glycoprotein is a glycosylphosphatidylinositol(GPI)-linked membrane protein carrying Dombrock blood group antigens. There are no standardized typing reagents for Do(a) or Do(b). We have developed ten different monoclonal antibodies(MoAbs) that are specific for Dombrock. The objectives of this study were to characterize these MoAbs serologically and determine the epitopes they recognize. MoAbs were generated by standard fusion methods. Mice were immunized with transfected human embryonic kidney 293T cells expressing high levels Do(a) or Do(b). The MoAbs were tested serologically with untreated and enzymatically or chemically modified red blood cells (RBCs).Serologic inhibition studies were performed with synthetic peptides corresponding to Do(a) and Do(b) amino acid sequences.Pepscan epitope analysis was done on an array of immobilized tridecapeptides corresponding to the full-length polypeptide. All ten antibodies were serologically specific for Dombrock. Eight of the antibodies recognized epitopes that were resistant to treatment with ficin, pronase, a-chymotrypsin, and neuraminidase,but sensitive to trypsin and 0.2 M dithiothreitol (DTT). Five have anti-Do(b)-like specificity. The epitope recognized by MIMA-52 was neuraminidase sensitive, and MIMA-127 epitope recognized a DTT-resistant, linear epitope (90)QKNYFRMWQK(99) of the Dombrock polypeptide. MIMA-127 was the only one of the ten Dombrock MoAbs mapped to a specific sequence of the Dombrock glycoprotein; the other nine MoAbs did not provide aspecific peptide binding pattern. The other MoAbs could not be mapped as they most likely recognize nonlinear, conformation-dependent epitopes, as is evident by their sensitivity to reduction of disulfide bonds by DTT. The dependence of some epitopes on antigen glycosylation is also a possibility.

Red cells from the original JAL+ proband are also DAK+ and STEM+.

BACKGROUND: The low-prevalence Rh antigen, JAL, was named after the index case, Mr. J. Allen. Based on reactivity of seven multi-specific sera with his RBCs, it was apparent that they express at least one additional low-prevalence antigen. The purpose of this study was to investigate the other low-prevalence antigen(s) on J. Allen's RBCs. METHODS: Blood samples and reagents were from our collections. Hemagglutination and DNA analyses were performed by standard methods. RESULTS: Our DNA analyses confirmed the presence of RHCE*ceS(340T) in J. Allen and revealed the presence of RHCE*ceBI (ce 48C, 712G, 818T, 1132G) and RHD*DOL (509T, 667T). RBCs from J. Allen were agglutinated by anti-JAL, anti-STEM, and anti-DAK. Two of the reactive multi-specific sera reported in the original paper reacted with RBCs from J. Allen, and with RBCs from four other people with RHCE*ceBI, including the original STEM+ index case (P. Stemper) but not with RBCs with the DIIIa, DAK+ phenotype. We conclude that they contain anti-STEM. CONCLUSION: J.Allen's RBCs express the low-prevalence Rh antigens, JAL, V/VS (extremely weakly), STEM, and DAK. The presence of JAL on the variant Rhce, RhceJAL (16Cys, 114Trp, 245Val), STEM on the variant Rhce, RhceBI (16Cys, 238Val, 273Val, 378Val), and DAK on the variant RhD (170Thr, 223Val), encoded by RHD*DOL in trans to RHCE*ceBI is consistent with expression of these antigens. When J. Allen RBCs are used to detect and identify an anti-JAL, it is important to remember that they also express STEM and DAK.

Penile cancer in Jamaicans managed at the University Hospital of the West Indies.

OBJECTIVES: The aim of this study is to determine the prevalence and clinicopathological correlates of penile cancer as well as the clinical outcomes in a sample of Jamaicans managed at the University Hospital of the West Indies (UHWI). METHODS: All available records of patients diagnosed with penile cancer from 1998-2008 at the UHWI were obtained. Patient demographics, circumcision status, sexually transmitted infection status, lesion duration, location and size, and lymph node status were obtained. Histology, differentiation and stage were recorded. Information was obtained regarding treatment and outcome. The current data were compared with a previous report from UHWI in 1959. RESULTS: The records of 22 of 26 patients with penile cancer were available for review. Mean (SD) age of patients was 68 (13) years. Eighteen (86%) patients were uncircumcised Mean tumour size was 5.7 (2.6) cm; 8 (36%) lesions involved the entire penis. Sixteen (73%) lesions had clinically regional disease and 11 (52%) patients had advanced pathological stage. Surgical treatment was performed in 15 (68%) patients. Case fatality was 38%, with median survival following surgical intervention of 38 person-months. The major predictor of death in this series was increasing age (HR = 1.06, 95% CI 0.99, 1.1, p = 0.079). There was an increase in age and clinical stage of the cancer at presentation in the current series; however there was no difference in survival. CONCLUSION: Penile cancer is an uncommon cancer, seen at an advanced stage in Jamaicans. Overall survival is poor and advanced age is a major predictor of death.

The efficacy of calcitriol therapy in the management of bone loss and fractures: a qualitative review.

UNLABELLED: Osteoporosis, a skeletal disorder characterized by a reduction in bone strength, increases fracture risk. Primary osteoporosis is usually a result of reduced bone mineral density as a consequence of natural aging. Secondary osteoporosis is usually a result of a disease, such as cystic fibrosis, or medical treatment, such as corticosteroids or cancer treatment. INTRODUCTION: Currently, ten million Americans are osteoporotic and an additional 34 million have the precursor condition, osteopenia. Osteoporosis leads to 1.5 million fractures and 500,000 hospitalizations annually. Osteoporosis-related fractures increase mortality and reduce quality of life. Calcitriol, the active form of vitamin D, regulates intestinal calcium absorption, among other actions. During the past four decades, many clinical trials investigating the effect of calcitriol on bone loss have been performed. METHODS: We conducted a systematic qualitative review of clinical trials that assessed calcitriol for the treatment of osteoporosis and bone loss. In these clinical trials, calcitriol was used as a monotherapy and in combination with other therapeutic bone agents. RESULTS AND CONCLUSION: Studies using calcitriol monotherapy, although not conclusive, found that calcitriol slowed the rate of bone loss in a variety of populations. Calcitriol in combination with other therapeutic bone agents was shown to have additional bone-preserving effects when compared to the use of therapeutic bone agents alone. A common side-effect of calcitriol therapy was hypercalcemia and hypercalciuria, but the degree of hypercalcemia was mild. Recent research found that intermittent dosing can reduce hypercalcemia rates. Calcitriol, alone or in combination with other agents, should be considered for the therapy of osteoporosis.

The Gerbich blood group system: a review.

Antigens in the Gebrich blood group system are expressed on glycophorin C (GPC) and glycophorin D (GPD), which are both encoded by a single gene, GYPC. The GYPC gene is located on the long arm of chromosome 2, and Gebrich antigens are inherited as autosomal dominant traits. There are 11 antigens in the Gebrich blood group system, six of high prevalence (Ge2, Ge3, Ge4, GEPL [Ge10*], GEAT [Ge11*], GETI [Ge12*]) and five of low prevalence (Wb [Ge5], Ls(a) [Ge6], An(a) [Ge7], Dh(a) [Ge8], GEIS [Ge9]). GPC and GPD interact with protein 4.1R, contributing stability to RBC membrane. Reduced levels of GPC and GPD are associated with hereditary elliptocytosis, and Gebrich antigens act as receptors for the malarial parasite Plasmodium falciparum. Anti-Ge2 and anti-Ge3 have caused hemolytic transfusion reactions, and anti-Ge3 has produced hemolytic disease of the fetus and newborn (HDFN).

The Dombrock blood group system: a review.

The Dombrock blood group system (Do) consists of two antithetical antigens (Do(a) and Do(b)) and five antigens of high prevalence (Gy(a), Hy, Jo(a), DOYA, and DOMR). Do antigens are carried on the Dombrock glycoprotein, which is attached to the RBC membrane via glycosylphosphatidylinositol linkage. The gene (DO, ART4) encoding the Do glycoprotein, located on the short arm of chromosome 12, has been cloned and sequenced, allowing the molecular basis of the various Do phenotypes to be determined. Do(a) and Do(b) have a prevalence that makes them useful as genetic markers; however, the paucity of reliable anti-Do(a) and anti-Do(b) has prevented this potential from being realized. The ease with which these antigens can be predicted by analysis of DNA opens the door for such studies to be carried out. Anti-Do(a) and anti-Do(b) are rarely found as a single specificity, but they have been implicated in causing hemolytic transfusion reactions. This review is a synthesis of our current knowledge of he Dombrock blood group system.

RHCE*ceAR encodes a partial c (RH4) antigen.

Thr Rh blood group system is highly complex both in the number of discreet antigens and in the existence of partial antigens, especially D and e. Recently, several partial c antigens have been reported. Here we report findings on an African American man with sickle cell disease whose RBCs typed C+c+ and whose plasma contained anti-c. Hemagglutination tests, DNA extraction, PCR-RFLP, reticulocyte RNA isolation, RT-PCR cDNA analyses, cloning, and sequencing were performed by standard procedures. RBCs from the patient typed C+c+ but his plasma contained alloanti-c. DNA analyses showed the presence of RHCE*Ce in trans to RHCE*ceAR with RHD*D and RHD*Weak D type 4.2.2. The amino acid changes on RhceAR are such that C+c+ patient made alloanti-c. This case shows that RhceAR carries a partial c antigen and illustrates the value of DNA testing as an adjunct to hemagglutination to aid in antibody identification in unusual cases.

Consortium for Blood Group Genes (CBGG): 2009 report.

Consortium for Blood Group Genes is a worldwide organization whose goal is to have a vehicle to interact, establish guidelines, operate a proficiency program, and provide education for laboratories involved in DNA and RNA testing for the prediction of blood group, platelet, and neutrophil antigens. Currently, the consortium operates with representatives from Brazil, Canada, and the United States. Membership is voluntary with the expectation that members actively contribute to discussions involving blood group genetics. This year witnessed a change in the standing committee membership and the institution of a representative for the human platelet antigens group. Looking forward, the consortium sees challenges for the nomenclature of blood group alleles and user-required specifications for laboratory information systems to store genotype information.

"It has to speak to people's everyday life...": qualitative study of men and women's willingness to participate in a non-medical approach to Chlamydia trachomatis screening.

OBJECTIVE: To explore the factors associated with men and women's willingness to provide a urine sample for Chlamydia trachomatis screening in various non-medical settings. METHODS: Men and women aged 16-24 years attending non-medical settings were invited to participate in urine-based screening and later to participate in a follow-up in-depth interview. Participant observation techniques were also used to collect data on young people's response to the offer of screening. RESULTS: The views of 24 men and women revealed three themes in relation to willingness to participate, particularly among men: their raised awareness of chlamydia, particularly its asymptomatic nature; the convenience of the offer; and the "non-medical" nature of the screening. In contrast, women more often felt the public nature of the settings inhibited them from agreeing to take the test and, thus, acted as a barrier to their willingness to participate in screening. CONCLUSIONS: The gender difference in willingness to participate in non-medical screening suggests that extending the reach of screening could certainly assist in bringing more young men into screening but may not necessarily destigmatise screening for women. As such, the potential benefits to men must be considered in the context of the potential psychosocial harms to women.

Signal transduction by glycophorin A: role of extracellular and cytoplasmic domains in a modulatable process.

Binding of ligands to the extracellular region of the erythrocyte transmembrane protein glycophorin A induces a decrease in membrane deformability. Since the property of membrane deformability is regulated by the skeletal proteins on the cytoplasmic side of the membrane, this suggests that ligand binding may initiate a transmembrane signal. To further study this process, we examined which domains of the extracellular region of glycophorin are involved in signal transduction and whether the cytoplasmic domain of the molecule is necessary for transmitting the signal. Using the ektacytometer, we compared the effect on deformability of four monoclonal antibodies that detect different epitopes on glycophorin A. We found that 9A3 (which recognized the amino terminus of glycophorin) caused a 5.8-fold increase in rigidity, R-10 and 10F7 (which recognized epitopes in the mid-region of the extracellular domain) caused a 10.8-fold increase in rigidity and B14 (which binds to glycophorin close to the membrane) caused a 18-fold increase in rigidity. Further, a direct relationship was observed between the degree of antibody-induced rigidity and the amount of glycophorin A that became associated with the skeletal proteins in a Triton shell assay. In Miltenberger V erythrocytes, which contain a hybrid sialoglycoprotein with no cytoplasmic domain, antibody binding did not induce an increase in rigidity. These results imply that glycophorin A is capable of a modulatable form of transmembrane signaling that is determined by the extracellular domain to which the ligand binds, and the cytoplasmic domain of glycophorin A is crucial for this process.

Acute sensitivity of landslide rates to initial soil porosity.

Some landslides move imperceptibly downslope, whereas others accelerate catastrophically. Experimental landslides triggered by rising pore water pressure moved at sharply contrasting rates due to small differences in initial porosity. Wet sandy soil with porosity of about 0.5 contracted during slope failure, partially liquefied, and accelerated within 1 second to speeds over 1 meter per second. The same soil with porosity of about 0.4 dilated during failure and slipped episodically at rates averaging 0.002 meter per second. Repeated slip episodes were induced by gradually rising pore water pressure and were arrested by pore dilation and attendant pore pressure decline.

Characterising transmission of a tuberculosis genotype in Scotland: a qualitative approach to social network enquiry.

SETTING: Contact investigation resulting from specimens sent to the Scottish Mycobacteria Reference Laboratory. OBJECTIVE: To characterise patients and types of exposures associated with transmission of a prevalent Mycobacterium tuberculosis genotype in Scotland. DESIGN: A combined approach using molecular epidemiology and semi-structured patient interviews for social network enquiry. RESULTS: We investigated social connections between 64 patients diagnosed between 1994 and 2004. Fifty-five per cent had > or = 1 identifiable contact. One third (n = 14, 32.6%) of the 43 epidemiological links detected were discerned as a result of patient interviews and were not previously recorded on surveillance reports, nor recognised by nurse specialists (all were non-household contacts). Sixteen putative sites of exposure were identified, 11 were public houses. Rather than a single-source outbreak, eight pockets of transmission were identified, the largest involving UK-born alcohol-misusing males frequenting several public houses. CONCLUSIONS: Using a standardised approach to explore themes around which individuals may have been exposed to TB resulted in the detection of previously unrecognised epidemiological links. Epidemiological data obtained from cluster investigations, e.g., risk and social behaviours that increase the risk of infection and sites of putative exposure, can enhance the development of more appropriate questions for the contact tracing interview.

Alloanti-c in a c-positive, JAL-positive patient.

BACKGROUND AND OBJECTIVES: In the Rh blood group system, partial D, C, and e antigens are well-known, but a partial c antigen resulting in the production of alloanti-c in a c+ individual is rare. One example of an alloanti-c in a c+ person was an anti-Rh26, which can appear as anti-c, and another was an alloanti-c in a c+ person with a presumed R(1)r phenotype. The finding of an apparent alloanti-c in a transfused c+ patient initiated this investigation. MATERIALS AND METHODS: Haemagglutination tests, DNA extraction, polymerase chain reaction (PCR)-based assays (PCR-restriction fragment length polymorphism, allele-specific PCR), reticulocyte mRNA extraction, reverse transcriptase (RT)-PCR and sequencing were performed by standard procedures. RESULTS: Plasma from a 64-year-old African American woman with a wound infection following a mastectomy contained anti-E, anti-S, anti-K, anti-Fy(a) and anti-Jk(b), reacting by the indirect antiglobulin test. In addition, the patient's plasma gave reactions that were consistent with an anti-c, while her pre-transfusion red blood cells typed c+ with some anti-c reagents. These results are consistent with a partial c antigen. The patient's red blood cells also typed V+(W)VS- and JAL+. Analyses of DNA and Rh-transcripts from this patient showed the presence of the following genes: RHD*D, RHD*DAU0, RHCE*Ce and RHCE*ce(S)(340). CONCLUSION: The nucleotide 340C>T change in RHCE exon 3 (predicted to encode 114Trp) of the RHCE*ce(S)(340) allele is associated with a JAL+ phenotype and the altered expression of the c, V and VS antigens. This alteration in the c antigen allowed the patient to make an alloanti-c. This case reveals that the RHCE*ce(S)(340) allele encodes a partial c antigen.

MNS blood group system: a review.

The MNS blood group system is second only to the Rh blood group system in its complexity. Many alloantibodies to antigens in the MNS system are not generally clinically significant although antibodies to low-prevalence and high-prevalence MNS antigens have caused hemolytic disease of the fetus and newborn. The MNS antigens are carried on glycophorin A (GPA), glycophorin B (GPB), or hybrids thereof, which arise from single-nucleotide substitution, unequal crossing over, or gene conversion between the glycophorin genes. Antigens in the MNS system are fully developed at birth. This review summarizes aspects of the MNS system, including the molecular basis of some antigens in the MNS blood group system. Readers are referred to existing excellent reviews for background information. Throughout this document, information given without references can be found in the reviews listed previously, and the reader is referred to these reviews for references to original reports.

Consortium for Blood Group Genes (CBGG): 2008 report.

The Consortium for Blood Group Genes is a worldwide organization whose goal is to have a vehicle to interact, establish guidelines, operate a proficiency program, and provide education for laboratories involved in DNA and RNA testing for the prediction of blood group, platelet, and neutrophil antigens.