The Glucatell (1r3)-b-D-glucan (BG) detection assay (Associates of Cape Cod) was studied as a diagnostic adjunct for invasive fungal infections (IFIs). On the basis of findings from a preliminary study of 30 candidemic subjects and 30 healthy adults, a serum BG level of у60 pg/mL was chosen as the cutoff. Testing was performed with serial serum samples obtained from 283 subjects with acute myeloid leukemia or myelodysplastic syndrome who were receiving antifungal prophylaxis. At least 1 serum sample was positive for BG at a median of 10 days before the clinical diagnosis in 100% of subjects with a proven or probable IFI. IFIs included candidiasis, fusariosis, trichosporonosis, and aspergillosis. Absence of a positive BG finding had a 100% negative predictive value, and the specificity of the test was 90% for a single positive test result and у96% for у2 sequential positive results. The Glucatell serum BG detection assay is highly sensitive and specific as a diagnostic adjunct for IFI.The mortality rate for invasive fungal infections in neutropenic subjects is 50% for subjects with Candida infection [1,2] and may approach 100% for those with invasive aspergillosis [3,4], fusariosis [5], or trichosporonosis [6]. Early diagnosis of invasive fungal infection in neutropenic subjects has the potential to increase antifungal therapeutic response, but meaningful diagnostic tests have proven to be elusive. Histopathologic demonstration of organisms in tissue specimens or growth of fungal agents in culture media is still the

A B S T R A C T PurposeThe current classification systems of myelodysplastic syndromes (MDS), including the International Prognostic Scoring System (IPSS), do not fully reflect the molecular heterogeneity of the disease. Molecular characterization may predict clinical outcome and help stratify patients for targeted therapies. Epigenetic therapy using decitabine, a DNA hypomethylating agent, is clinically effective for the treatment of MDS. Therefore, we investigated the association between DNA methylation and clinical outcome in MDS. Patients and MethodsWe screened 24 patients with MDS for promoter CpG island methylation of 24 genes and identified aberrant hypermethylation at 10 genes. We then performed quantitative methylation analyses by bisulfite pyrosequencing of the identified genes in 317 patient samples from three independent studies and assessed relations between methylation and clinical outcome. ResultsIn an initial training cohort of 89 patients with MDS, methylation frequencies of individual genes ranged from 7% to 70% and were highly concordant. Therefore, we defined a methylation z score based on all genes for each patient. We found that patients with higher levels of methylation, compared with patients with lower levels, had a shorter median overall survival (12.3 v 17.5 months, respectively; P ϭ .04) and shorter median progression-free survival (6.4 v 14.9 months, respectively; P ϭ .009). This methylation prognostic model was independent of age, sex, and IPSS group. Applied to two validation cohorts (228 patients), this model was confirmed as an independent prognostic predictor for survival. Although methylation at baseline did not correlate with clinical response to decitabine, we observed a significant correlation between reduced methylation over time and clinical responses. ConclusionDNA methylation predicts overall and progression-free survival in MDS. J Clin

A dults with relapsed/refractory acute lymphoblastic leukemia have an unfavourable prognosis, which is influenced by disease and patient characteristics. To further evaluate these characteristics, a retrospective analysis of 1,706 adult patients with Ph-negative relapsed/refractory B-precursor acute lymphoblastic leukemia diagnosed between 1990-2013 was conducted using data reflecting the standard of care from 11 study groups and large centers in Europe and the United States. Outcomes included complete remission, overall survival, and realization of stem cell transplantation after salvage treatment. The overall complete remission rate after first salvage was 40%, ranging from 35%-41% across disease status categories (primary refractory, relapsed with or without prior transplant), and was lower after second (21%) and third or greater (11%) salvage. The overall complete remission rate was higher for patients diagnosed from 2005 onward (45%, 95% CI: 39%-50%). One-and three-year survival rates after first, second, and third or greater salvage were 26% and 11%, 18% and 6%, and 15% and 4%, respectively, and rates were 2%-5% higher among patients diagnosed from 2005. Prognostic factors included younger age, longer duration of first remission, and lower white blood cell counts at primary diagnosis. This large dataset can provide detailed reference outcomes for patients with relapsed/refractory Ph-negative B-precursor acute lymphoblastic leukemia. clinicaltrials.gov identifier: 02003612 IntroductionOverall prognosis among adult acute lymphoblastic leukemia (ALL) patients has improved by optimisation of front-line therapy, 1 but outcomes remain poor for patients who relapse or are refractory to initial treatment. Reported rates of complete remission (CR) after salvage treatment range from 18%-45%, and median survival times range from 2-8 months, with less than 10% survival after 5 years in most studies.2-7 Achievement of CR and subsequent HSCT is the only curative option in relapsed adult ALL; 2,3,6 however, this can only be achieved in a subgroup For these reasons, new treatment options are needed for adult patients with relapsed/refractory (r/r) ALL, and a number of promising compounds are currently under clinical evaluation. 8,9 The rarity of adult r/r ALL, 10 combined with the very poor outcomes and non-standardised approaches of salvage therapies, make it difficult to conduct randomised trials of new compounds. Approvals of new treatments are therefore often based on evidence from phase 2 single-arm trials. 11-14To attain a more precise estimate of clinical practice outcomes in adult r/r ALL, and to evaluate important patient subgroups, we pooled data from major national study groups and large individual sites treating adult ALL from Europe and the United States to create the most extensive clinical dataset available in this population.The analysis aimed to describe patient characteristics and outcome parameters (achievement of CR, overall survival [OS] and realization of allogeneic HSCT) among adult patients with P...

We conducted this study to determine the feasibility and safety of cladribine followed by rituximab in patients with hairy cell leukemia including the variant form (HCLv). Cladribine 5.6 mg/m 2 given IV over 2 hours daily for 5 days was followed ϳ 1 month later with rituximab 375 mg/m 2 IV weekly for 8 weeks.Responses were recorded and BM minimal residual disease (MRD) was evaluated after the completion of rituximab. Thirty-six patients have been treated in- IntroductionOver the past quarter century, we have witnessed remarkable progress in the treatment of patients with hairy cell leukemia (HCL). 1,2 The nucleoside analogs cladribine and pentostatin have made a significant impact in the treatment of HCL producing complete response (CR) rates of 80% to 90%. 3,4 Long-term follow-up of patients treated with these agents have shown that about one-fourth to one-third of patients relapse after a follow-up of 3 to 4 years with little difference between the 2 drugs, in terms of durability of response. [5][6][7] Previous reports have suggested that the quality of the initial response can be predictive of the outcome, with a longer disease-free survival in those achieving a CR after initial therapy as opposed to those with lesser responses. [7][8][9] This has led to the suggestion by some authors to persist with therapy, in the absence of toxicity, until a CR is attained, 7 and to efforts to eradicate minimal residual disease (MRD) after the initial therapy. This is particularly important as the median age of patients with HCL is in the 50s, and subsequent CRs may be shorter in duration with successive therapy. 5,10,11 Determination of the risk of relapse based on the persistence of MRD determined by immunohistochemistry (IHC) using anti-CD45RO, anti-CD20, and DBA.44 in paraffin-embedded BM sections was first reported by the group from Northwestern University. 12 More recently, more precise methods using immunophenotyping by flow cytometry, and consensus primer or clonespecific PCR analysis of Ig receptors, have been used to detect MRD in patients with HCL. [13][14][15] Whether eradication of MRD should be a goal of therapy in the initial management of patients with HCL remains a subject of discussion. 16 Sigal et al have reported that among 19 patients with HCL in continuous CR after 1 course of cladribine (median time from therapy, 16 years, [range, 11-21 years]) in whom BM samples were available, 7 (37%) had MRD, and 3 (16%) had morphologic evidence of HCL suggesting that patients with MRD or even morphologic disease can live many years without a hematologic relapse. 17 We have previously reported on the efficacy of rituximab in eradicating MRD in patients with HCL treated with cladribine. 14 Among the 13 patients, reported MRD assessed by consensus primer PCR or flow cytometry was eradicated in 92% at 3 months. We have reported a brief summary of this phase 2 trial including patients with relapsed disease recently for a collective report of a meeting of hairy cell leukemia experts at the National Institutes of Heal...

The coronavirus disease 2019 (COVID-19) pandemic poses several challenges to the management of patients with leukemia. The biology of each leukemia and its corresponding treatment with conventional intensive chemotherapy, with or without targeted therapies (venetoclax, FLT3 inhibitors, IDH1/2 inhibitors, Bruton's tyrosine kinase inhibitors), introduce additional layers of complexity during COVID-19 highrisk periods. The knowledge about COVID-19 is accumulating rapidly. An important distinction is the prevalence of "exposure" versus "clinical infectivity," which determine the risk versus benefit of modifying potentially highly curative therapies in leukemia. At present, the rate of clinical infection is < 1-2% worldwide. With a mortality rate of 1-5% in CO-VID-19 patients in the general population and potentially of > 30% in patients with cancer, careful consideration should be given to the risk of COVID-19 in leukemia. Instead of reducing patient access to specialized cancer centers and modifying therapies to ones with unproven curative benefit, there is more rationale for less intensive, yet effective therapies that may require fewer clinic visits or hospitalizations. Here, we offer recommendations on the optimization of leukemia management during high-risk COVID-19 periods.

In the past few years, our improved knowledge of acute myeloid leukemia (AML) pathogenesis has led to the accelerated discovery of new drugs and the development of innovative therapeutic approaches. The role of the immune system in AML development, growth and recurrence has gained increasing interest. A better understanding of immunological escape and systemic tolerance induced by AML blasts has been achieved. The extraordinary successes of immune therapies that harness the power of T cells in solid tumors and certain hematological malignancies have provided new stimuli in this area of research. Accordingly, major efforts have been made to develop immune therapies for the treatment of AML patients. The persistence of leukemia stem cells, representing the most relevant cause of relapse, even after allogeneic stem cell transplant (allo-SCT), remains a major hurdle in the path to cure for AML patients. Several clinical trials with immune-based therapies are currently ongoing in the frontline, relapsed/refractory, post-allo-SCT and minimal residual disease/maintenance setting, with the aim to improve survival of AML patients. This review summarizes the available data with immune-based therapeutic modalities such as monoclonal antibodies (naked and conjugated), T cell engagers, adoptive T-cell therapy, adoptive-NK therapy, checkpoint blockade via PD-1/PD-L1, CTLA4, TIM3 and macrophage checkpoint blockade via the CD47/SIRPa axis, and leukemia vaccines. Combining clinical results with biological immunological findings, possibly coupled with the discovery of biomarkers predictive for response, will hopefully allow us to determine the best approaches to immunotherapy in AML.

In a seminal paper, Pakula and Sauer (Nature, 1990, 344, 363–364) demonstrated that the increase in side‐chain hydrophobicity has a reverse relationship with protein stability. We have addressed this problem with several examples of mutants that span at different locations in protein structure based on secondary structure and solvent accessibility. We confirmed that the stability change upon single coil mutation at exposed region is reversely correlated with hydrophobicity with a single exception. In addition, we found the existence of such relationship in partially buried coil mutants. The stability of exposed helical mutants is governed by conformational properties. In buried and partially buried helical and strand mutants properties reflecting hydrophobicity have direct relationship with stability, whereas an opposite relationship was obtained with entropy and flexibility. The structural analysis of partially buried/exposed mutants showed that the surrounding residues are important for the stability change upon mutation. These results provide insights to understand the general behavior for the stability of proteins upon amino acid substitutions. © 2009 Wiley Periodicals, Inc. Biopolymers 91: 591–599, 2009. This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com

Background The novel bispecific ligand-directed toxin (BLT) DT2219 consists of a recombinant fusion between the catalytic and translocation enhancing domain of diphtheria toxin (DT) and bispecific single chain variable fragments (scFV) of antibodies targeting human CD19 and CD22. We conducted a phase 1 dose escalation study to assess the safety, maximum tolerated dose (MTD), and preliminary efficacy of DT2219 in patients with relapsed/refractory B cell lymphoma or leukemia. Methods DT2219 was administered intravenously over 2 hours every other day for 4 total doses. Dose was escalated from 0.5 μg/kg/day to 80 μg/kg/day in nine dose cohorts until a dose limiting toxicity (DLT) was observed. Results Twenty-five patients with mature or precursor B-cell lymphoid malignancies expressing CD19 and/or CD22 enrolled to the study. Patients received median 3 prior lines of chemotherapy and 8 failed hematopoietic transplantation. All patients received a single course of DT2219; one patient was retreated. The most common adverse events (AE) including, weight gain, low albumin, transaminitis and fevers were transient grade 1-2 and occurred in patients in higher dose cohorts (≥40 μg/kg/day). Two subjects experienced DLT at dose levels 40 and 60 μg/kg. Durable objective responses occurred in 2 patients; one was complete remission after 2 cycles. Correlative studies showed a surprisingly low incidence of neutralizating antibody (30%). Conclusions We have determined the safety of a novel immunotoxin DT2219 and established it's biologically active dose between 40-80 μg/kg/day ×4. A phase II study exploring repetitive courses of DT2219 is planned.