Laboratory and Clinical Implications of Incidental and Secondary Germline Findings During Tumor Testing.

CONTEXT.­: Next-generation sequencing is a powerful clinical tool for cancer management but can produce incidental/secondary findings that require special consideration. OBJECTIVE.­: To discuss clinical and laboratory issues related to incidental or secondary germline findings in the clinical setting of tumor testing and inform future guidelines in this area. DESIGN.­: A College of American Pathologists workgroup including representation from the American Society of Clinical Oncology, the Association for Molecular Pathology, and the American College of Medical Genetics and Genomics created a review of items that should be considered when developing guidelines for incidental or secondary findings when performing clinical tumor testing. RESULTS.­: Testing recommendations should be cognizant of the differences among anticipated incidental, unanticipated incidental, and secondary findings, and whether normal tissue is also tested. In addition to defining which variants will be reported, robust recommendations must also take into account test design and validation, reimbursement, cost, infrastructure, impact on reflex testing, and maintenance of proficiency. Care providers need to consider the potential of a test to uncover incidental or secondary findings, the recommendation of upfront counseling, the need for consent, the timing of testing and counseling, and that the exact significance of a finding may not be clear. CONCLUSIONS.­: As clinical oncology testing panels have become a mainstay of clinical cancer care, guidelines addressing the unique aspects of incidental and secondary findings in oncology testing are needed. This paper highlights clinical and laboratory considerations with regard to incidental/secondary findings and is a clarion call to create recommendations.

Will pathogen reduction of blood components harm more people than it helps in developed countries?

BACKGROUND: Blood-borne infectious diseases are a major impediment to the provision of safe blood. Pathogen reduction (PR) technologies have been approved for the treatment of plasma and platelet (PLT) concentrates to reduce infectious complications and graft-versus-host disease but product potency is adversely affected STUDY DESIGN AND METHODS: We reviewed published data describing PR technology for estimates of treated blood component physical and functional loss. These physical and functional losses were summed and projected onto measured effects of plasma and PLT dose in trauma resuscitation. The net benefits estimated as reduced infectious disease deaths were compared to net losses estimated as increased deaths from uncontrolled hemorrhage. RESULTS: Transfusion-transmitted infectious diseases caused five or fewer acute deaths each year from 2009 through 2014 in the United States according to the Food and Drug Administration. In-hospital deaths from uncontrolled hemorrhage after trauma number more than 10,000 yearly and are reduced by 4% to 15% with concentrated blood product resuscitation. The loss of 20% of plasma potency and 30% of PLT potency to PR is likely to be associated with 400 extra trauma deaths each year. Trauma represents a small fraction, perhaps 15%, of all massively transfused individuals. CONCLUSIONS: Resuscitation of massive hemorrhage may be limited by blood component potency as shown in our literature review and analysis. The safety-versus-potency trade involved with current blood plasma and PLT PR technology is likely to result in a net loss of life. Hemorrhagic risk from reduced blood product potency for patients with trauma and other indications for massive transfusion is an important consideration in risk-based decision making for implementing PR.

Rapid HIV Testing in a New Orleans Emergency Department is Effective in Identifying New HIV Diagnoses and in Linking Patients to Care.

A retrospective chart review of patients who agreed to a rapid HIV test in the emergency department in the initial year of institution of the rapid test was conducted. Out of 8,204 patients, 99 were newly diagnosed with HIV in the first year of the institution of the rapid HIV test (1.2%). Eighty-five (86%) had a documented referral to the infectious disease clinic, and 59 (60%) were linked to care within one year of diagnosis. The majority (58%) of the patients with a new diagnosis of HIV had been seen in the Interim Louisiana State University Public Hospital (ILPH) healthcare system in the five years prior to their diagnosis. Forty-nine percent of the patients met diagnostic criteria of AIDS at diagnosis. Rapid HIV testing in the emergency department is an effective way to find previously undiagnosed patients and link them to subspecialty care.

Better hemostatic profiles of never-frozen liquid plasma compared with thawed fresh frozen plasma.

BACKGROUND: Immediate use of thawed fresh frozen plasma (FFP) when resuscitating hemorrhagic shock patients has become more common. According to the AABB (formerly known as American Association of Blood Banks), FFP is the preferred product that can be used up to 5 days after thawing. However, limited data exist on the clinical use and hemostatic profiles of Food and Drug Administration-approved liquid plasma (LQP), which can be stored at 1 °C to 6 °C for up to 26 days. We characterized changes in LQP hemostatic potential during 26 days of cold storage. METHODS: Ten FFP and 10 LQP single-donor units, matched by sex and blood group, were analyzed. FFP was thawed and kept refrigerated for 5 days and LQP for 26 days. Plasma samples were evaluated at Days 0 and 5 for thawed plasma (TP) and 0, 5, 10, 20, and 26 for LQP, by thrombelastography, thrombogram, platelet counts, platelet microparticles, clotting factors, and natural coagulation inhibitors. RESULTS: LQP had a better capacity to form a clot and generate thrombin compared with TP. LQP's hemostatic potential, expressed as endogenous thrombin potential (total amount of thrombin [nM] formed over time [minute]), initially exceeded that of TP (1,425 vs. 1,184, p < 0.05) but decreased to levels similar to TP by Day 26 (1,201 vs. 1,103, p = 0.15). Significantly higher platelet microparticles were found in LQP on Day 26 compared with those in LQP on Day 0 (23.6 x 10(9)/L vs. 3 x 10(9)/L, p < 0.001) or those in TP on Day 5 (2.8 x 10(9)/L). By Day 26, the majority of clotting factors and inhibitors retained more than 88% of their initial activities in LQP, with the few exceptions of factors well known to be unstable. CONCLUSION: The hemostatic profiles of LQP were better and sustained five times longer than the more commonly used TP, indicating that never-frozen plasma can be considered for use in the United States in trauma patients requiring immediate plasma resuscitation.

Pathology case of the month. Swamp-related wound infection. Aeromonas hydrophila.

A 38 year-old woman with no significant past medical history was brought to the emergency department following a motor vehicle rollover and submersion into swampy waters alongside a Louisiana interstate. In addition to multiple blunt force injuries, she suffered a laceration injury over the right lower extremity, which initially required irrigation and sterile dressing. On hospital day two, the wound became grossly infected. The wound was swabbed for culture and Gram stain, and the patient was empirically started on intravenous Piperacillin/tazobactam. Results from the Gram stain showed few white blood cells and numerous Gram negative rods. The following day, the wound continued to drain purulent material but with a stable zone of erythema. The wound was drained and debrided. On hospital day four, the blood agar plate, previously inoculated with the wound culture, grew the colonies shown in the below photograph.

Damage control resuscitation: from emergency department to the operating room.

Damage control surgery emphasizes limited operations with control of bleeding and contamination. Traditional management centered upon correction of acidosis and hypotension with crystalloids. Damage control resuscitation (DCR) is permissive hypotension and early hemostatic resuscitation combined identified and corrects coagulopathy with fresh-frozen plasma (FFP), restricting use of crystalloids. We hypothesize a survival advantage in patients managed with DCR when compared with a historical cohort of patients. During the 2-year retrospective review, a 1-year period after institution of DCR was compared with a historical control. Resuscitation strategies were analyzed and stratified into emergency department (ED) resuscitation and intraoperative resuscitation. Univariate analysis of continuous data was done with Student's t test followed by multiple logistic regression. Fifty-seven and 61 patients were managed during the NonDCR and DCR periods respectively. Baseline demographic patient characteristics and physiologic variables were similar between groups. ED DCR patients received less crystalloids: 1.1 versus 4.7 liters (P = 0.0001), more FFP: 1.8 versus 0.5 (P = 0.001). NonDCR had a lower initial systolic pressure in the operating room when compared with DCR: 81 mm Hg versus 95 mm Hg (P = 0.03). DCR patients received less intraoperative crystalloids: 5.7 versus 15.8 liters (P = 0.0001) and more FFP: 15.1 versus 6.2 (P = 0.0001). DCR conveyed a survival benefit (Odds Ratio; 95% confidence interval: 0.40 (0.18-0.90), P = 0.024). NonDCR group had 13.2 days longer hospital length of stay. Damage control resuscitation, beginning in the ED, used more packed red blood cells and FFP minimizing crystalloids. DCR was associated with a survival advantage and shorter length of stay in patients with severe hemorrhage.

Damage control resuscitation in combination with damage control laparotomy: a survival advantage.

BACKGROUND: Damage control laparotomy (DCL) improves outcomes when used in patients with severe hemorrhage. Correction of coagulopathy with close ratio resuscitation while limiting crystalloid forms a new methodology known as damage control resuscitation (DCR). We hypothesize a survival advantage in DCL patients managed with DCR when compared with DCL patients managed with conventional resuscitation efforts (CRE). METHODS: This study is a 4-year retrospective study of all DCL patients who required >or=10 units of packed red blood cells (PRBC) during surgery. A 2-year period after institution of DCR (DCL and DCR) was compared with the preceding 2 years (DCL and CRE). Univariate analysis of continuous data was done with Student's t test followed by multiple logistic regression. RESULTS: One Hundred twenty-four and 72 patients were managed during the DCL and CRE and DCL and DCR time periods, respectively. Baseline patient characteristics of age, Injury Severity Score, % penetrating, blood pressure, hemoglobin, base deficit, and INR were similar between groups. There was no difference in quantity of intraoperative PRBC utilization between DCL and CRE and DCL and DCR study periods: 21.7 units versus 25.5 units (p = 0.53); however, when compared with DCL and CRE group, patients in the DCL and DCR group received less intraoperative crystalloids, 4.7 L versus 14.2 L (p = 0.009); more fresh frozen plasma (FFP), 18.2 versus 6.4 (p = 0.002); a closer FFP to PRBC ratio, 1 to 1.2 versus 1 to 4.2 (p = 0.002); platelets to PRBC ratio, 1:2.3 versus 1:5.9 (0.002); shorter mean trauma intensive care unit length of stay, 11 days versus 20 days (p = 0.01); and greater 30-day survival, 73.6% versus 54.8% (p < 0.009). The addition of DCR to DCL conveyed a survival benefit (odds ratio; 95% confidence interval: 0.19 (0.05-0.33), p = 0.005). CONCLUSION: This is the first civilian study that analyses the impact of DCR in patients managed with DCL. During the DCL and DCR study period more PRBC, FFP, and platelets with less crystalloid solution was used intraoperatively. DCL and DCR were associated with a survival advantage and shorter trauma intensive care unit length of stay in patients with severe hemorrhage when compared with DCL and CRE.

Hemostatic resuscitation during surgery improves survival in patients with traumatic-induced coagulopathy.

BACKGROUND: Although hemostatic resuscitation with a 1:1 ratio of fresh-frozen plasma (FFP) to packed red blood cells (PRBC) after severe hemorrhage has been shown to improve survival, its benefit in patients with traumatic-induced coagulopathy (TIC) after >10 units of PRBC during operation has not been elucidated. We hypothesized that a survival benefit would occur when early hemostatic resuscitation was used intraoperatively after injury in patients with TIC. METHODS: A 7-year retrospective study of patients with emergency department diagnosis of TIC after transfusion of >10 units of PRBC in the operating room. TIC was defined as initial emergency department international normalized ratio > 1.2, prothrombin time > 16 seconds, and partial thromboplastin time > 50 seconds. Patients were divided into FFP:PRBC ratios of 1:1, 1:2, 1:3, and 1:4. Patients with diagnosis of TIC who received transfusion of both FFP and PRBC during surgery were included. Other variables evaluated included age, gender, mechanism of injury, initial base deficit, mean operative time, trauma intensive care unit length of stay (TICU LOS) and Injury Severity Score. The primary outcome measure evaluated was the impact of the early FFP:PRBC ratio on mortality. RESULTS: Four hundred thirty-five patients underwent emergency operations postinjury and received FFP with >10 units of PRBC in the operating room; 135 (31.0%) of these patients had TIC and 53 died (39.5% mortality). Mean operative time was 137 minutes (SD +/- 49). There were no differences with regard to age, gender, mechanism of injury, initial base deficit, or Injury Severity Score among all groups. A significant difference in mortality was found in patients who received >10 units of PRBC when FFP:PRBC ratio was 1:1 versus 1:4 (28.2% vs. 51.1%, p = 0.03). Intermediate mortality rates were noted in patients with 1:2 and 1:3 ratios (38% and 40%, respectively). From a linear regression model, 13 days of increased TICU LOS was observed among 1:4 group compared with 1:1 group (p < 0.01). CONCLUSION: TIC is common after severe injury and is associated with a high mortality in patients transfused with >10 units of PRBC during surgery. Early hemostatic resuscitation during first hours after injury improves survival with shorter TICU LOS in patients with TIC.

Current evidence based guidelines for factor VIIa use in trauma: the good, the bad, and the ugly.

Recombinant factor VII (rFVIIa) has arisen as an option for the control of life-threatening traumatic bleeding unresponsive to other means. The timing of administration, dosage, mortality, units of blood transfusion saved, risk of thrombotic events, and risk/benefits ratio are presently poorly defined. A Medline search from 1995 through March 2008 was conducted. All English language articles containing the terms "trauma" and "factor VII" or its variants were retrieved. Letters to the editor, animal studies, and general reviews were excluded. A total of 19 articles met inclusion criteria. These articles were then reviewed and stratified into three classes of evidence according to the quality assessment instrument developed by the Brain and Trauma Foundation. Levels of recommendation were developed. A total of 118 articles were identified. Only one Class I study was identified. This study demonstrated that three doses of rFVIIa given in blunt traumatic hemorrhage yielded a significant reduction of 2.6 of red blood cells used. These findings were not statistically significant for penetrating trauma patients. There was no reduction in mortality and no increase in thromboembolic events. Four Class II studies were identified; three showed a significant decrease of blood product usage and one demonstrated significant reductions in 24-hour and 30 day death from hemorrhage in patients receiving rFVIIa. The remaining 14 studies were Class III reviews of databases, registries, case series, and case reports. No identified study specifically addressed the cost/benefit analysis of rFVIIa usage in trauma hemorrhage. Utility of rFVIIa in trauma-associated hemorrhage remains controversial. There is Level I supporting the use of rFVIIa for blunt trauma patients only. There is no Class I evidence supporting decreased mortality or differences in thromboembolic events. Minimal effective dosing regimens and cost/benefit analyses have not yet been examined.

Performance characteristics of a quantitative TaqMan hepatitis C virus RNA analyte-specific reagent.

We determined the dynamic range, reproducibility, accuracy, genotype bias, and sensitivity of the TaqMan hepatitis C virus (HCV) analyte-specific reagent (ASR). Serum samples were processed using the MagNA Pure LC instrument and run on the COBAS TaqMan 48 analyzer. The performance characteristics of the ASR were also compared with those of the qualitative AMPLICOR and quantitative AMPLICOR MONITOR HCV tests. The ASR exhibited a >/=6-log(10) linear dynamic range and excellent reproducibility, with a mean coefficient of variation of 14%. HCV RNA concentration measured with the ASR agreed within an average of 0.42 log(10) (2.6-fold) of the labeled concentration with members of a standard reference panel. HCV genotypes 1 to 4 were amplified with similar efficiencies with the ASR. The ASR and AMPLICOR MONITOR viral load results were significantly correlated (r = 0.8898; P < 0.01), but the agreement was poor (mean difference, 0.45 +/- 0.35 log(10)) for 72 HCV RNA-positive clinical samples. However, 98.9% agreement between the ASR and qualitative AMPLICOR test results was found with 60 positive and 29 negative samples. Limiting-dilution experiments demonstrated that the limits of detection for ASR and AMPLICOR tests were 84 and 26 IU/ml, respectively. The performance characteristics of the TaqMan HCV ASR are appropriate for all clinical applications of HCV RNA testing.