Roles of the C-terminal domains of topoisomerase IIα and topoisomerase IIß in regulation of the decatenation checkpoint.

Topoisomerase (topo) IIα and IIß maintain genome stability and are targets for anti-tumor drugs. In this study, we demonstrate that the decatenation checkpoint is regulated, not only by topo IIα, as previously reported, but also by topo IIß. The decatenation checkpoint is most efficient when both isoforms are present. Regulation of this checkpoint and sensitivity to topo II-targeted drugs is influenced by the C-terminal domain (CTD) of the topo II isoforms and by a conserved non-catalytic tyrosine, Y640 in topo IIα and Y656 in topo IIß. Deletion of most of the CTD of topo IIα, while preserving the nuclear localization signal (NLS), enhances the decatenation checkpoint and sensitivity to topo II-targeted drugs. In contrast, deletion of most of the CTD of topo IIß, while preserving the NLS, and mutation of Y640 in topo IIα and Y656 in topo IIß inhibits these activities. Structural studies suggest that the differential impact of the CTD on topo IIα and topo IIß function may be due to differences in CTD charge distribution and differential alignment of the CTD with reference to transport DNA. Together these results suggest that topo IIα and topo IIß cooperate to maintain genome stability, which may be distinctly modulated by their CTDs.

Accreditation of Individualized Quality Control Plans by the College of American Pathologists.

The Laboratory Accreditation Program of the College of American Pathologists (CAP) began in 2015 to allow accredited laboratories to devise their own strategies for quality control of laboratory testing. Participants now have the option to implement individualized quality control plans (IQCPs). Only nonwaived testing that features an internal control (built-in, electronic, or procedural) is eligible for IQCP accreditation. The accreditation checklists that detail the requirements have been peer-reviewed by content experts on CAP's scientific resource committees and by a panel of accreditation participants. Training and communication have been key to the successful introduction of the new IQCP requirements.

College of American Pathologists' laboratory standards for next-generation sequencing clinical tests.

CONTEXT: The higher throughput and lower per-base cost of next-generation sequencing (NGS) as compared to Sanger sequencing has led to its rapid adoption in clinical testing. The number of laboratories offering NGS-based tests has also grown considerably in the past few years, despite the fact that specific Clinical Laboratory Improvement Amendments of 1988/College of American Pathologists (CAP) laboratory standards had not yet been developed to regulate this technology. OBJECTIVE: To develop a checklist for clinical testing using NGS technology that sets standards for the analytic wet bench process and for bioinformatics or "dry bench" analyses. As NGS-based clinical tests are new to diagnostic testing and are of much greater complexity than traditional Sanger sequencing-based tests, there is an urgent need to develop new regulatory standards for laboratories offering these tests. DESIGN: To develop the necessary regulatory framework for NGS and to facilitate appropriate adoption of this technology for clinical testing, CAP formed a committee in 2011, the NGS Work Group, to deliberate upon the contents to be included in the checklist. Results . -A total of 18 laboratory accreditation checklist requirements for the analytic wet bench process and bioinformatics analysis processes have been included within CAP's molecular pathology checklist (MOL). CONCLUSIONS: This report describes the important issues considered by the CAP committee during the development of the new checklist requirements, which address documentation, validation, quality assurance, confirmatory testing, exception logs, monitoring of upgrades, variant interpretation and reporting, incidental findings, data storage, version traceability, and data transfer confidentiality.

Blood storage duration and biochemical recurrence of cancer after radical prostatectomy.

OBJECTIVE: To test the hypothesis that perioperative transfusion of allogeneic and autologous red blood cells (RBCs) stored for a prolonged period speeds biochemical recurrence of prostate cancer after prostatectomy. PATIENTS AND METHODS: We evaluated biochemical prostate cancer recurrence in men who had undergone radical prostatectomy and perioperative blood transfusions from July 6, 1998, through December 27, 2007. Those who received allogeneic blood transfusions were assigned to nonoverlapping "younger," "middle," and "older" RBC storage duration groups. Those who received autologous RBC transfusions were analyzed using the maximum storage duration as the primary exposure. We evaluated the association between RBC storage duration and biochemical recurrence using multivariable Cox proportional hazards regression. RESULTS: A total of 405 patients received allogeneic transfusions. At 5 years, the biochemical recurrence-free survival rate was 74%, 71%, and 76% for patients who received younger, middle, and older RBCs, respectively; our Cox model indicated no significant differences in biochemical recurrence rates between the groups (P=.82; Wald test). Among patients who received autologous transfusions (n=350), maximum RBC age was not significantly associated with biochemical cancer recurrence (P=.95). At 5 years, the biochemical recurrence-free survival rate was 85% and 81% for patients who received younger and older than 21-day-old RBCs, respectively. CONCLUSION: In patients undergoing radical prostatectomy who require RBC transfusion, recurrence risk does not appear to be independently associated with blood storage duration.

Prognosticators in thigh soft tissue sarcomas.

BACKGROUND: In sarcoma patients the roles of smoking history, family cancer history, and leukoreduced blood transfusions have not been studied and the effect of preoperative radiation on blood loss has not been examined. METHODS: Seventy-seven patients with non-metastatic and non-recurrent thigh sarcomas surgically treated at the Cleveland Clinic were identified. Among patient variables studied were: close family history of cancer, perioperative transfusion history, smoking history, and radiation history. Median follow-up for the survivors was 3.2 years. RESULTS: We found that tumor grade, transfusion >3 U (P = 0.022), and pre- or post-operative radiation therapy (P = 0.041) were risk factors for distant metastasis. Tumor grade (P = 0.008), positive smoking history (P = 0.039), and >3 U of non-leukoreduced blood transfused (P = 0.037) were risk factors for death of any-cause. Close family history of cancer correlated with having a grade 3 sarcoma (P = 0.044). Neoadjuvant radiotherapy correlated with >3 U of blood transfused (P = 0.001) and biopsy performed at the treating institution led to a significant decrease in rate of recurrence (P = 0.016). CONCLUSIONS: We present novel findings in terms of transfusions, family history of cancer and site of initial biopsy in sarcoma patients.

Laboratory standardization from the patient's point of view.

Optimal patient care is best defined in terms of outcome. Reliable laboratory test results are important to patient safety. The laboratory must use the tools available to it to minimize the uncertainty of measurement. Three sources of error contribute to uncertainty: Intermethod bias, which is minimized and trueness of measure maximized when laboratories use calibrators and methods traceable to higher order, reference standards; imprecision inherent in the analysis, which is seen as small differences between replicate tests; interference from factors external to the test itself, which are seen as erroneous values markedly deviant from trueness. Although improbable, such contributions to total analytical error may be the most misleading. Risk is best managed by identifying the sources of error and controlling for those sources most likely to contribute to total analytical error. Comprehensive control of error requires the laboratory scientist and physicians caring for patients to work together to ensure interpretability of results. Practice guidelines are available from the Clinical and Laboratory Standards Institute to address each of these factors.

Duration of red-cell storage and complications after cardiac surgery.

BACKGROUND: Stored red cells undergo progressive structural and functional changes over time. We tested the hypothesis that serious complications and mortality after cardiac surgery are increased when transfused red cells are stored for more than 2 weeks. METHODS: We examined data from patients given red-cell transfusions during coronary-artery bypass grafting, heart-valve surgery, or both between June 30, 1998, and January 30, 2006. A total of 2872 patients received 8802 units of blood that had been stored for 14 days or less ("newer blood"), and 3130 patients received 10,782 units of blood that had been stored for more than 14 days ("older blood"). Multivariable logistic regression with propensity-score methods was used to examine the effect of the duration of storage on outcomes. Survival was estimated by the Kaplan-Meier method and Blackstone's decomposition method. RESULTS: The median duration of storage was 11 days for newer blood and 20 days for older blood. Patients who were given older units had higher rates of in-hospital mortality (2.8% vs. 1.7%, P=0.004), intubation beyond 72 hours (9.7% vs. 5.6%, P<0.001), renal failure (2.7% vs. 1.6%, P=0.003), and sepsis or septicemia (4.0% vs. 2.8%, P=0.01). A composite of complications was more common in patients given older blood (25.9% vs. 22.4%, P=0.001). Similarly, older blood was associated with an increase in the risk-adjusted rate of the composite outcome (P=0.03). At 1 year, mortality was significantly less in patients given newer blood (7.4% vs. 11.0%, P<0.001). CONCLUSIONS: In patients undergoing cardiac surgery, transfusion of red cells that had been stored for more than 2 weeks was associated with a significantly increased risk of postoperative complications as well as reduced short-term and long-term survival.

Frequencies and characterization of cytogenetically unrelated clones in various hematologic malignancies: seven years of experiences in a single institution.

Cytogenetically unrelated clones are uncommon findings in hematologic disorders. In the present study, among the 1,110 various hematologic malignancies analyzed during the past seven years, 27 (2.4%) patients had karyotypically unrelated clones: 3.5% (7/202) of acute myeloid leukemias, 5.3% (11/206) of myelodysplastic syndromes, none of 40 acute lymphoblastic leukemias, 0.4% (1/233) of myeloproliferative disorders, and 2.6% (8/306) lymphoproliferative disorders with clonal chromosomal abnormalities. Twenty-five patients showed two unrelated clones and two had three unrelated clones. The most consistent chromosome abnormalities were del(5q) (seven cases), +8 (six cases), del(20q) (five cases), and del(7q), +12, +21, and -22 (three cases each). Of interest, the high frequency of different numeric or structural abnormalities affecting the same chromosomes in such clones supports the hypothesis that these karyotypically unrelated clones originate from the common malignant clone through submicroscopic molecular genetic changes and evolutionary processes.

Acute myocardial infarction in the hospital and delay in blood transfusion / Infarto agudo do miocárdio e sua relação com o retardo transfusional

Peer reviews of hospital transfusion practices rarely screen for under-transfusion, one definition of which is a delay in transfusion that complicates significant myocardial ischemia. A one-year, retrospective review studied the timing of red cell transfusions among medical and surgical patients with secondary diagnoses of acute myocardial infarction. Five possible cases of delayed transfusion were found among 11,197 transfused patients (incidence = 1 in 2,200 transfusions). Delayed transfusion can complicate a patient's difficult clinical course. Hospital transfusion committees should consider surveying for delayed transfusion in addition to the more common surveys for over-transfusion.

Os revisores de prática transfusional hospitalar, raramente avaliOs revisores de prática transfusional hospitalar, raramente avaliam a subindicação transfusional, que propiciam retardo na transfusão possível de complicação isquêmica miocárdica. Um estudo retrospectivo de um ano de indicações de concentrado de glóbulo entre pacientes clínicos e cirúrgicos com diagnóstico secundário de infarto agudo do miocárdio foi realizado. Cinco casos de possível retardo na indicação transfusional foram observados em 11.197 pacientes transfundidos (incidência = 1 em 2.200 transfusões). O retardo na indicação de transfusões pode complicar pacientes em estado grave, sugerindo que os comitês transfusionais devem ficar atentos para o fato, e não só para os casos de excesso de transfusões.

Detection and correction of systematic laboratory problems by analysis of clustered proficiency testing failures.

CONTEXT: The Laboratory Accreditation Program of the College of American Pathologists monitors the performance of its subscribers in proficiency testing (PT). Failure to perform as expected prompts the program to query the laboratory. OBJECTIVE: To determine whether laboratories are correcting apparent problems when contacted by the program about repeatedly unacceptable performance in a diagnostic test. DESIGN: Retrospective analysis of 1 year's records (2002-2003) from the College's Proficiency Testing Exception Summary correspondence, which identifies clusters of PT failures. The analysis focused on those laboratories in which the Proficiency Testing Exception Summary algorithm identified repeated failures over 3 or 4 testing events; PT performance is monitored as a condition of accreditation. During 1 survey year, approximately 6300 accredited laboratories collectively tested approximately 1,205,000 analytes and submitted results to their PT providers on more than 3,500,000 PT challenges. During the period of observation, 14,085 Proficiency Testing Exception Summary reports were mailed to participants. Educational materials were included to help laboratories identify and correct each PT failure. RESULTS: There were only 1304 cases of repeated PT failures after the initial correspondence from the accreditation program (9.3%). Of these, there were only 119 cases of unsatisfactory results on the subsequent PT event (9.1%). All systematic problems were resolved by the conclusion of the third round of correspondence. CONCLUSIONS: Accredited laboratories generally perform well in proficiency testing. Identification of clusters of PT failures by the accreditation provider can help those laboratories having analytic difficulties to investigate and correct the problems.

The cost of patient safety.

To ensure patient safety, it costs an organization time, money, and commitment. The clinical laboratory may promote patient safety or may contribute to medical error. Laboratory errors put a patient at risk at any point along the path of workflow. The cost of an initiative in patient safety may be considered from four perspectives: compliance, feasibility, present risk, and financial. Three examples are offered to illustrate the use of these approaches. Most patient-safety strategies in laboratory medicine are not expensive. They are affordable with a structured outlay of existing resources and a willingness to follow defined work practices without exception. More extensive projects, especially those that cross jurisdictional lines within an organization, do require comprehensive project management. Management ofboth kinds of initiatives is addressed by NCCLS' documents on quality practice.

A case of fatal West Nile virus meningoencephalitis associated with receipt of blood transfusions after open heart surgery.

First identified in the United States in 1999, West Nile virus caused approximately 3,500 infections in the late summer and fall of 2002. The virus is predominantly transmitted by mosquitoes, and the risk of infection through blood product transfusion is believed to be low. We present a case of West Nile virus encephalitis transmitted by red blood cell transfusion at the time of coronary artery bypass grafting that resulted in the patient's death. Individuals undergoing procedures with high blood product transfusion requirements, such as cardiac surgery or organ transplantation, may be at higher risk of this nosocomial infection during epidemics.

A pulmonary syndrome in patients with acute myelomonocytic leukemia and inversion of chromosome 16.

Different subtypes of acute myelogenous leukemia have distinct clinical presentations and courses. The specific clinical and molecular aspects of these leukemias have helped modify and create specific strategies for their management. We observed an increased incidence of pulmonary complications in patients with acute myelomonocytic leukemias (AMML) with inversion of chromosome 16 [inv(16)] irrespective of the presence of hyperleukocytosis. We reviewed patient records available over a period of 12 years at The Cleveland Clinic Foundation of patients with AMML with inv(16) and compared the incidence of pulmonary complications to a matched control group of patients with AMML but without inv(16). We found an increased incidence of pulmonary complications in the AMML with inv(16)group when compared to the control group. Two of these patients demonstrated brochiolitis obliterans with organizing pneumonia (BOOP) on lung biopsy. No specific etiology for the pulmonary complications was identified. These findings represent the first observation of an association between WHO-AMML with inv(16) [FAB-AML M4 with inv(16)] with a pulmonary syndrome at presentation. BOOP should be suspected in these cases. A larger prospective study to evaluate this association is warranted.

Allergic transfusion reactions: an evaluation of 273 consecutive reactions.

CONTEXT: Allergic reaction to transfusion is common. However, the review of a large series of allergic transfusion reactions has not been performed. OBJECTIVE: To review a large series of allergic transfusion reactions. DESIGN: A retrospective review of all reported and evaluated transfusion reactions during a 9-year period at 1 institution was performed. Associated clinical signs and symptoms were evaluated. SETTING: Large, tertiary-care teaching hospital. RESULTS: A total of 1613 adverse reactions to transfusion were evaluated. Allergic transfusion reactions accounted for 17% (273 of 1613) of the transfusion reactions. Severe allergic reactions (anaphylaxis, anaphylactoid signs and symptoms, and/or hypotension) were observed in 21 patients (7.7% of allergic reactions, or 1.3% of all transfusion reactions). Serum tryptase, a marker for anaphylaxis, was measured in 1 patient and determined to be borderline elevated. Five patients experienced allergic transfusion reactions to autologous red cell transfusions. One patient experienced hives during the transfusion of a major ABO mismatched red blood cell. A wide variety of skin manifestations were observed, but 26 (9.5%) patients did not have skin manifestations. Allergic transfusion reactions were estimated to occur in approximately 1 in 4124 blood components transfused, or 1 in 2338 transfusion episodes. Severe allergic reactions occurred in approximately 1 in 30,281 transfusions. No deaths directly attributable to transfusion were observed in this patient group. CONCLUSIONS: The clinical presentation of allergic transfusion reactions was quite variable, and the pathophysiology remains unclear. Recommendations for clinical evaluation and therapy remain problematic and often empirical.