Two novel antithetical KN blood group antigens may contribute to more than a quarter of all KN antisera in Europe.

BACKGROUND: All antigens described in the KN blood group system are located in the long homologous repeat D (LHR-D) of complement receptor 1 (CR1). While there have been reports that some sera react only with the long homologous repeat C (LHR-C), the antigens in LHR-C are unknown. STUDY DESIGN AND METHODS: Recombinant LHR-C and LHR-D were used to identify antibodies directed against LHR-C of CR1, into which a point mutation was introduced to characterize the underlying blood group antigens. In addition, database studies to define haplotypes of CR1 were performed. RESULTS: Several antisera were identified that were specific against CR1 p.1208His and against CR1 p.1208Arg, located in LHR-C. Fifteen KN haplotypes were found in the Ensembl genome browser. It was shown that due to a linkage disequilibrium anti-CR1 p.1208His may be mistaken for anti-KCAM. CONCLUSION: A novel antithetical KN blood group antigen pair was found at position p.1208 of CR1, for which the names DACY and YCAD are proposed. Antibodies against these two novel antigens seem to contribute to more than a quarter of all KN sera in Europe.

The panacea statistical toolbox of a biomedical peer reviewer.

The main role of a peer reviewer is to make judgments on the research articles by asking a number of questions to evaluate the quality of the research article. Statistics is a major part of any biomedical research article, and most reviewers gain their experiences in manuscript reviewing by undertaking it but not through an educational process. Therefore, reviewers of the biomedical journals normally do not have enough knowledge and skills to evaluate the validity of statistical methods used in biomedical research articles submitted for consideration. Hence, inappropriate statistical analysis in medical journals can lead to misleading conclusions and incorrect results. In this paper, the most common basic statistical guidelines are described that might be a road map to the biomedical reviewers. It is not meant for statisticians or medical editors who have special interest and expertise in statistical analysis.

Secreted ß3-integrin enhances natural killer cell activity against acute myeloid leukemia cells.

Integrins are a large family of heterodimeric proteins that are involved in cell adhesion, migration, and proliferation. Integrin diversity and function is regulated by alternative splicing. Membrane-bound and truncated ß3-integrins were shown to be key players in cancer metastasis. However, the immunomodulatory functions of the soluble (s) ß3-integrin have not been investigated yet. In this study, we described a novel form of sß3-integrin in acute myeloid leukaemia (AML) patients. Furthermore, we assessed the role of the sß3-integrin in the modulation of natural killer (NK)-cell activity. Levels of sß3-integrin were analysed in plasma samples of 23 AML patients and 26 healthy donors by ELISA. The capacity of sß3-integrin to regulate NK cell activity was investigated using proliferation, cytokine secretion, and cytotoxicity assays. Circulating sß3-integrin was detected in the plasma of 8 AML patients. NK cells showed significantly higher proliferation rates after stimulation with sß3-integrin and IL-2, IL-15 (73%). Significant increases in the NK cells' secreted levels of TNF-α, IFN-γ were measured in presence of sß3-integrin. In addition, sß3-integrin caused the upregulation of Granzyme B transcripts levels as well as FasL expression levels in NK cells. Most importantly, significantly higher K562 or AML blast target cell lysis rates were observed when NK cells were exposed to sß3-integrin. This study reports the identification of a novel sß3-integrin in AML patients and provides novel insights into its role in the immunomodulation of NK cell activity.

The panacea toolbox of a PhD biomedical student.

Doing a PhD (doctor of philosophy) for the sake of contribution to knowledge should give the student an immense enthusiasm through the PhD period. It is the time in one's life that one spends to "hit the nail on the head" in a specific area and topic of interest. A PhD consists mostly of hard work and tenacity; however, luck and genius might also play a little role. You can pass all PhD phases without having both luck and genius. The PhD student should have pre-PhD and PhD toolboxes, which are "sine quibus non" for getting successfully a PhD degree. In this manuscript, the toolboxes of the PhD student are discussed.

Medical literature review: Search or perish.

Literature review is a cascading process of searching, reading, analyzing, and summing up of the materials about a specific topic. However, searching the literature is like searching "a needle in a haystack", and hence has been called "Cinderella".(1) Therefore, skills and effective pathways of searching the literature are needed to achieve high sensitive and specific results.

Development of a single-antigen magnetic bead assay (SAMBA) for the sensitive detection of HPA-1a alloantibodies using tag-engineered recombinant soluble ß3 integrin.

Timely and accurate testing for human platelet antigen 1a (HPA-1a) alloantibodies is vital for clinical diagnosis of neonatal alloimmune thrombocytopenia (NAIT). Current antigen-specific assays used for the detection of HPA-1 alloantibodies are technically very complex and cumbersome for most diagnostic laboratories. Hence, we designed and applied recombinant soluble (rs) ß3 integrins displaying HPA-1a or HPA-1b epitopes for the development of a single-antigen magnetic bead assay (SAMBA). Soluble HPA-1a and HPA-1b were produced recombinantly in human embryonic kidney 293 (HEK293) cells and differentially tagged. The recombinant soluble proteins were then immobilized onto paramagnetic beads and used for analysis of HPA-1 alloantibodies by enzyme-linked immunosorbent assay (ELISA). HPA-1a serum samples (n=7) from NAIT patients, inert sera and sera containing non-HPA-1a antibodies were used to evaluate the sensitivity and specificity of the SAMBA. Fusion of V5-His or GS-SBP-His tags to the rsß3 integrins resulted in high-yield expression. SAMBA was able to detect all HPA-1a and -1b alloantibodies recognized by monoclonal antibody-specific immobilization of platelet antigens assay (MAIPA). No cross-reactions between the sera were observed. Two out of seven of the HPA-1a alloantibody-containing sera demonstrated weak to moderate reactivity in MAIPA but strong signals in SAMBA. SAMBA provides a very reliable method for the detection of HPA-1 antibodies with high specificity and sensitivity. This simple and rapid assay can be adapted for use in any routine laboratory and can be potentially adapted for use on automated systems.

The Rh allele frequencies in Gaza city in Palestine.

BACKGROUND: The Rh blood group system is the second most clinically significant blood group system. It includes 49 antigens, but only five (D, C, E, c and e) are the most routinely identified due to their unique relation to hemolytic disease of the newborn (HDN) and transfusion reactions. Frequency of the Rh alleles showed variation, with regard to race and ethnic. OBJECTIVES: The purpose of the study was to document the Rh alleles' frequencies amongst males (M) and females (F) in Gaza city in Palestine. MATERIALS AND METHODS: Two hundred and thirty-two blood samples (110 M and 122 F) were tested against monoclonal IgM anti-C,anti-c, anti-E, anti-e and a blend of monoclonal/polyclonal IgM/IgG anti-D. The expected Rh phenotypes were calculated using gene counting method. RESULTS: The most frequent Rh antigen in the total sample was e, while the least frequent was E.The order of the combined Rh allele frequencies in both M and F was CDe > cDe > cde > CdE > cDE > Cde > CDE. A significant difference was reported between M and F regarding the phenotypic frequencies (P < 0.05). However, no significance (P > 0.05) was reported with reference to the observed and expected Rh phenotypic frequencies in either M or F students. CONCLUSION: It was concluded that the Rh antigens, alleles and phenotypes in Gaza city have unique frequencies, which may be of importance to the Blood Transfusion Center in Gaza city and anthropology.

Quality Indicators in Laboratory Medicine: from theory to practice. Preliminary data from the IFCC Working Group Project "Laboratory Errors and Patient Safety".

BACKGROUND: The adoption of Quality Indicators (QIs) has prompted the development of tools to measure and evaluate the quality and effectiveness of laboratory testing, first in the hospital setting and subsequently in ambulatory and other care settings. While Laboratory Medicine has an important role in the delivery of high-quality care, no consensus exists as yet on the use of QIs focussing on all steps of the laboratory total testing process (TTP), and further research in this area is required. METHODS: In order to reduce errors in laboratory testing, the IFCC Working Group on "Laboratory Errors and Patient Safety" (WG-LEPS) developed a series of Quality Indicators, specifically designed for clinical laboratories. In the first phase of the project, specific QIs for key processes of the TTP were identified, including all the pre-, intra- and post-analytic steps. The overall aim of the project is to create a common reporting system for clinical laboratories based on standardized data collection, and to define state-of-the-art and Quality Specifications (QSs) for each QI independent of: a) the size of organization and type of activities; b) the complexity of processes undertaken; and c) different degree of knowledge and ability of the staff. The aim of the present paper is to report the results collected from participating laboratories from February 2008 to December 2009 and to identify preliminary QSs. RESULTS AND CONCLUSIONS: The results demonstrate that a Model of Quality Indicators managed as an External Quality Assurance Program can serve as a tool to monitor and control the pre-, intra- and post-analytical activities. It might also allow clinical laboratories to identify risks that lead to errors resulting in patient harm: identification and design of practices that eliminate medical errors; the sharing of information and education of clinical and laboratory teams on practices that reduce or prevent errors; the monitoring and evaluation of improvement activities.