Northern lights assay: a versatile method for comprehensive detection of DNA damage.

To access publisher's full text version of this article, please click on the hyperlink in Additional Links field or click on the hyperlink at the top of the page marked DownloadDNA damage assays have various limitations in types of lesions detected, sensitivity, specificity and samples that can be analyzed. The Northern Lights Assay (NLA) is based on 2D Strandness-Dependent Electrophoresis (2D-SDE), a technique that separates nucleic acids based on length, strandness, structure and conformation changes induced by damage. NLA is run on a microgel platform in 20-25 min. Each specimen is analyzed in pairs of non-digested DNA to detect single- and double-stranded breaks (DSBs) and Mbo I-digested DNA to detect other lesions. We used NLA to evaluate DNA in solution and isolated from human cells treated with various genotoxic agents. NLA detected and distinguished between single- and DSBs, interstrand and intrastrand DNA crosslinks, and denatured single-stranded DNA. NLA was sufficiently sensitive to detect biologically relevant amount of DNA damage. NLA is a versatile, sensitive and simple method for comprehensive and simultaneous analysis of multiple types of damage, both in purified DNA and in DNA isolated from cells and body fluids. NLA can be used to evaluate DNA quality in biosamples, monitor complex molecular procedures, assess genotoxicity, diagnose genome instability, facilitate cancer theranostics and in basic nucleic acids research.University of Iceland Research Fund Landspitali University Hospital Research Fund Icelandic Center for Research Funds Lifeind ehf. University of Iceland Research Fun

Red blood cell alloimmunization in pregnancy during the years 1996-2015 in Iceland: a nation-wide population study

To access publisher's full text version of this article click on the hyperlink belowBACKGROUND: Red blood cell (RBC) alloimmunization during pregnancy is still a major problem. Historically, anti-D antibodies are most likely to cause severe hemolysis, but other antibodies are also important. In Iceland, postnatal RhIg prophylaxis was implemented in 1969, universal RBC antibody screening was implemented in 1978, but antenatal RhIg prophylaxis is not yet routine. STUDY DESIGN AND METHODS: This nation-wide population study gathered data on alloimmunized pregnancies in Iceland between 1996 and 2015. Blood bank alloimmunization data were linked to Icelandic Medical Birth Registry data. RBC antibodies were classified as either clinically significant or clinically nonsignificant. RESULTS: In total, 912 positive antibody screens from 87,437 births were identified (1.04% prevalence). The most frequent antibodies were anti-M (19.4%), anti-E (19.0%), and anti-D (12.5%). Anti-D prevalence among D-negative mothers was 1.1%. Icelandic Medical Birth Registry data were available for 881 (96.6%) pregnancies. In the clinically significant group (n = 474), anti-E (27%) and anti-D (20%) were most common, whereas anti-M was most frequent (53%) in the clinically nonsignificant group (n = 407). Mothers in the clinically significant group were older, more often multigravidae, had more abortions and stillbirths, and had shorter gestational length. Newborns in the clinically significant group were less healthy, had lower weight and Apgar scores, and required more treatment. Among specificities in the clinically significant group, anti-D antibodies were most strongly associated with severe hemolysis. CONCLUSION: In this study, the prevalence of alloimmunization was similar to that in previous reports. Of all clinically significant antibodies, anti-D was most strongly associated with severe hemolysis, requiring phototherapy or exchange transfusions. Our data emphasize the importance of implementing an antenatal prophylactic RhIg program in Iceland in the near future

Red blood cell utilization and transfusion triggers in patients diagnosed with chronic lymphocytic leukaemia in Iceland 2003-2016.

To access publisher's full text version of this article click on the hyperlink belowBACKGROUND AND OBJECTIVES: Revised Icelandic guidelines proposed a restrictive haemoglobin (Hb) threshold of 70 g/l for red blood cell (RBC) transfusions in general, but 100 g/l for malignancies/bone marrow suppression. Chronic lymphocytic leukaemia (CLL) is frequently complicated by anaemia. The objective was to investigate RBC transfusion practices in CLL. MATERIALS AND METHODS: This retrospective nation-wide study utilized an Icelandic registry of CLL patients diagnosed between 2003 and 2016. Medical records were reviewed and haemoglobin transfusion triggers compared for two periods: Earlier (2003-2012) and latter (2013-2017). RESULTS: Two hundred and thirteen patients were diagnosed with CLL over the period whereof 77 (36·2%) received RBC transfusion(s). Median time from diagnosis to first transfusion was 2·2 years. Higher age, Rai stage 3/4 at diagnosis (P < 0·05) and chemotherapy (P < 0·001) were associated with increased odds of transfusions. Shorter time to first transfusion correlated with higher age (P < 0·001) and Rai stage (P = 0·02) at diagnosis. The mean Hb trigger was 90·4 and 81·2 in the earlier and latter period respectively (P = 0·01). This difference in Hb triggers was most pronounced in patients without documented bone marrow involvement, or 80·5 g/l compared to 93·5 g/l (P = 0·004). The median time from diagnosis to transfusion was longer in the latter period (2·9 years vs. 1·6 years, P = 0·01). After RBC transfusions the survival decreased significantly (P < 0·001). CONCLUSION: One-third of CLL patients received RBC transfusions but few were heavily transfused. Older age, Rai stage, and chemotherapy predicted RBC use. The Hb transfusion trigger decreased over time while time to first RBC transfusion increased. RBC transfusions predict poor survival.Harold-Gunson foundatio

To Wash or Not to Wash? Comparison of Patient Outcome after Infusion of Cryopreserved Autologous Hematopoietic Stem Cells before and after the Replacement of Manual Washing by Bedside Thawing.

To access publisher's full text version of this article click on the hyperlink belowPrior to infusion, cryopreserved autologous peripheral blood stem cell (auto-PBSC) grafts can either be thawed at the bedside or thawed and washed at the laboratory. At our center, manual washing of grafts prior to infusion was discontinued in April 2012 and bedside thawing was implemented. This study compares the outcomes of two patient groups who received auto-PBSC either after post-thaw washing (n = 84) or bedside thawing (n = 83). No life-threatening infusion-related side effects were reported in either group. There was no significant difference in the mean CD34+ cells/kg dose of infused auto-PBSC in the two groups (p = 0.41), nor in the number of days to neutrophils > 0.5 × 109/L (p = 0.14), days to platelets > 20 × 109/L (p = 0.64), or days to platelets > 50 × 109/L (p = 0.62) after transplant. There was also no difference in the number of days on total parenteral nutrition (p = 0.69), days on G-CSF therapy (p = 0.48), or days with fever (p = 0.73). Finally, there was no significant difference in the number of red cell units transfused (p = 0.32), or platelet units transfused (p = 0.94) after the transplant. One-hundred-day mortality was identical in the two groups (2.4%). Both thawing procedures are safe and result in acceptable engraftment and patient outcomes

Noninvasive fetal RHD genotyping to guide targeted anti-D prophylaxis-an external quality assessment workshop.

To access publisher's full text version of this article click on the hyperlink belowBACKGROUND AND OBJECTIVES: Fetal RHD genotyping of cell-free fetal DNA from RhD-negative pregnant women can be used to guide targeted antenatal and postnatal anti-D prophylaxis for the prevention of RhD immunization. To assure the quality of clinical testing, we conducted an external quality assessment workshop with the participation of 28 laboratories. MATERIALS AND METHODS: Aliquots of pooled maternal plasma were sent to each laboratory. One sample was positive, and the second sample was negative for fetal RHD, verified by pre-workshop testing using quantitative real-time PCR (qPCR) analysis of RHD exons 4, 5, 7 and 10. Plasma samples were shipped at room temperature. A reporting scheme was supplied for data collection, including questions regarding the methodological setup, results and clinical recommendations. Different methodological approaches were used, all employing qPCR with a total of eight different combinations of RHD exon targets. The samples were tested blindly. RESULTS: Fetal RHD genotyping was performed with no false-negative and no false-positive results. One inconclusive result was reported for the RHD-positive sample, and four inconclusive results were reported for the RHD-negative sample. All clinical conclusions were satisfactory. CONCLUSION: This external quality assessment workshop demonstrates that despite the different approaches taken to perform the clinical assays, fetal RHD genotyping is a reliable laboratory assay to guide targeted use of Rh prophylaxis in a clinical setting