Reevaluating Patient Eligibility for Inotuzumab Ozogamicin Based on CD22 Expression: Is Dim Expression Sufficient?

Salvage options for patients with relapsed B-cell acute lymphoblastic leukemia (B-ALL) include inotuzumab ozogamicin (InO), a recombinant, humanized anti-CD22 monoclonal antibody conjugated to the cytotoxic antibiotic calicheamicin. However, the benefit of InO in patients with dim CD22 expression remains unclear. We present a case of a patient with B-ALL who responded to InO despite only dim surface expression of CD22 by flow cytometry, achieving a survival benefit concordant with that reported in the literature and maintaining a good quality of life as a transfusion-independent outpatient. Our observation has broad relevance to clinicians who manage patients with B-ALL who are candidates for InO.     

Evaluation of Cortactin and HS1 Genes Expression: New Players in Adult B-Cell Acute Lymphoblastic leukemia

Objectives: This study aimed to assess the prognostic value of cortactin and HS1 genes expression in adult B-cell acute lymphoblastic leukemia. Methods: The study included a cohort of 74 adult B-ALL patients and 76 controls. Cortactin and HS1 genes expression were quantified by real time PCR. Results: The expression of cortactin and HS1 were significantly higher in B-ALL patients at diagnosis as compared to post induction levels (p     

Viruses and cancers: possible role of hepatitis C virus

Oncogenesis is a multifactorial process in which environmental, genetic and infectious factors are variably involved. A possible role of specific viruses has been suggested in at least 15% of human cancers. Hepatitis C virus (HCV), which is both hepato- and lymphotropic, is responsible for various liver disorders, i.e. chronic hepatitis, cirrhosis and hepatocellular carcinoma, as well as for a constellation of extrahepatic immune-mediated manifestations, among which is mixed cryoglobulinaemia. This is a systemic disorder secondary to a chronic, benign B-lymphocyte proliferation, which in some subjects may evolve to a malignant non-Hodgkin's lymphoma (NHL). Interestingly, recent studies reported the appearance of malignant B-cell neoplasias in patients with type C chronic hepatitis; moreover, in a significant number (from 22% to 50%) of 'idiopathic' NHLs, the presence of HCV infection has been demonstrated. The presence of a geographical etherogeneity in the prevalence of HCV-positive NHL suggests that other co-factors, i.e. genetic and environmental, could be involved in the lymphomagenesis. HCV may exert its oncogenic potential in two different directions, leading to liver cancer or B-cell lymphoma.     

Transfusion patterns in pediatric open heart surgery

BACKGROUND: Transfusions in pediatric open heart surgery were analyzed to determine the percentage of patients transfused, the types and volumes of blood components used, and the relationships among transfusions, patient characteristics, surgeon, and surgical procedure. STUDY DESIGN and METHODS: In a 9-month period, 122 patients, aged 12 or less (median, 1 year; 31% <4 months old), underwent 126 procedures (37 routine, 60 complex, 29 repeat operations). Bypass circuit size and priming solution, target intraoperative hematocrit, heparinization, protamine reversal, and transfusion indications and doses were standardized. The number of full "adult" units of packed red cells (RBCs), units of fresh-frozen plasma (FFP), and platelet concentrates (PCs) transfused in the operating room through postoperative Day 3 were tabulated. RESULTS: RBCs, FFPs, and PCs were transfused in 98, 54, and 58 percent of cases, respectively. Twenty-two percent of components were transfused postoperatively. The average numbers of components transfused for complex procedures (3.4 RBCs, 6.1 total) and repeat operations (4.0 RBCs, 8.1 total) were greater than those for routine procedures (1.8 RBCs, 2.1 total) (p<0.01). The average total number of components transfused did not correlate with surgeon or patient age; patients <4 months old used the largest mean numbers of RBCs and total components of all types. For four procedures, preoperative crossmatch and directed-donation collection orders that would be expected to produce acceptable utilization rates and a <15-percent chance of needing additional components were determined. CONCLUSION: Blood order protocols for pediatric open heart surgery can be procedure-specific, should address the use of non-red cell components, and should cover early postoperative transfusions.     

Transforming growth factor-β1 and phosphatases modulate COX-2 protein expression and TAU phosphorylation in cultured immortalized podocytes

Objective and design

The aim of this study is to elucidate TGF-β1 signaling pathways involved in COX-2 protein induction and modulation of TAU protein phosphorylation in cultured podocytes.

Materials, treatment and methods

In vitro cultured immortalized podocytes were stimulated with TGF-β1 in presence and absence of pharmacologic inhibitors for various signaling pathways and phosphatases. Then, COX-2 protein expression, as well as P38MAPK, AKT and TAU phosphorylation levels were evaluated by western blot analysis.

Results

TGF-β1 induction of COX-2 protein levels was completely blocked by pharmacologic inhibitors of phosphatases, P38 MAPK, or NF-?B pathways. Time course experiments showed that TGF-β1 activated p38 MAPK after 5 min of stimulation. Interestingly, podocyte co-incubated with TGF-β1, high glucose and/or PGE2 showed strong increase in p38 MAPK and AKT phosphorylation as well as COX- 2 protein expression levels. Levels of phosphorylated AKT were further reduced and levels of phosphorylated p38 were increased when PGE2 was added to the culture media. Interestingly, selective phosphatases inhibitors completely abrogated PGE2-induced P38 MAPK and TAU phosphorylation. Also, inhibition of phosphatases reversed TGF-β1–induced COX-2 protein expression either alone or when incubated with high glucose or PGE2.

Conclusion

These data suggest TGF-β1 mediates its effect in podocyte through novel signaling mechanisms including phosphatases and TAU protein phosphorylation.

     

Evaluating the impact of in silico predictors on clinical variant classification

PurposeAccording to the American College of Medical Genetics and Genomics/Association of Medical Pathology (ACMG/AMP) guidelines, in silico evidence is applied at the supporting strength level for pathogenic (PP3) and benign (BP4) evidence. Although PP3 is commonly used, less is known about the effect of these criteria on variant classification outcomes.MethodsA total of 727 missense variants curated by Clinical Genome Resource expert groups were analyzed to determine how often PP3 and BP4 were applied and their impact on variant classification. The ACMG/AMP categorical system of variant classification was compared with a quantitative point-based system. The pathogenicity likelihood ratios of REVEL, VEST, FATHMM, and MPC were calibrated using a gold standard set of 237 pathogenic and benign variants (classified independent of the PP3/BP4 criteria).ResultsThe PP3 and BP4 criteria were applied by Variant Curation Expert Panels to 55% of missense variants. Application of those criteria changed the classification of 15% of missense variants for which either criterion was applied. The point-based system resolved borderline classifications. REVEL and VEST performed best at a strength level consistent with moderate evidence.ConclusionWe show that in silico criteria are commonly applied and often affect the final variant classifications. When appropriate thresholds for in silico predictors are established, our results show that PP3 and BP4 can be used at a moderate strength.     

Radionuclide quantification of mitral and aortic regurgitation

Abstract. Gated equilibrium bloodpool scintigraphy was used to obtain the ratio of left and right ventricular stroke counts (end-diastolic minus end-systolic counts within ventricular areas of interest), the radionuclide equivalent of stroke volumes. This ratio or stroke count index (SCI) should be unity in normal subjects and increased in patients with aortic or mitral regurgitation, when left ventricular stroke output rises to compensate for regurgitant flow. Results of this non-invasive method were compared with semiquantitative angiographic grading of mitral (1 to 4+) or aortic (1 to 3+) regurgitation in ninety-seven patients. We found a SCI of 1.15 ± 0.18 (SD) in thirty-six control subjects without evidence of mitral or aortic regurgitation at cardiac catheterization. Subsequently, a ratio of 1.50 was chosen as the upper limit of normal for the analysis of thirty-seven patients with mitral regurgitation and twenty-four patients with aortic regurgitation. Clearly, elevated SCI values were obtained in the presence of grade 3 and 4 mitral regurgitation (eighteen out of twenty patients) and of grade 2 and 3 aortic regurgitation (seventeen out of eighteen patients). Only two out of seventeen subjects with grade 1 or 2 mitral incompetence had an elevated SCI, while none of six subjects with grade I aortic regurgitation had an abnormal SCI. If these lesser degrees of valvular incompetence are considered of minor significance, overall sensitivity of the radionuclide method in our patient population was 92%, specificity 95%. Occasional discrepancies between SCI and angiographic severity of left-sided valvular regurgitation are probably a result of methodological limitations. We could not demonstrate any relation with global left ventricular function as measured from the radionuclide ejection fraction. We conclude that the SCI may be used as a non-invasive tool for diagnosis and management of patients with valvular heart disease, both before and after interventions.