Chronic Lymphocytic Leukemia/Small Lymphocytic Lymphoma, Version 4.2020, NCCN Clinical Practice Guidelines in Oncology.

Chronic lymphocytic leukemia (CLL) and small lymphocytic lymphoma (SLL) are characterized by a progressive accumulation of leukemic cells in the peripheral blood, bone marrow, and lymphoid tissues. Treatment of CLL/SLL has evolved significantly in recent years because of the improved understanding of the disease biology and the development of novel targeted therapies. In patients with indications for initiating treatment, the selection of treatment should be based on the disease stage, patient's age and overall fitness (performance status and comorbid conditions), and cytogenetic abnormalities. This manuscript discusses the recommendations outlined in the NCCN Guidelines for the diagnosis and management of patients with CLL/SLL.

NCCN Guidelines Insights: Chronic Lymphocytic Leukemia/Small Lymphocytic Lymphoma, Version 2.2019.

Chronic lymphocytic leukemia (CLL) is generally characterized by an indolent disease course. Histologic transformation (also known as Richter's transformation) to more aggressive lymphomas, such as diffuse large B-cell lymphoma or Hodgkin lymphoma, occurs in approximately 2% to 10% of patients and is associated with a poor prognosis. These NCCN Guidelines Insights discuss the recommendations for the diagnosis and management of patients with histologic transformation.

Hairy Cell Leukemia, Version 2.2018, NCCN Clinical Practice Guidelines in Oncology.

Hairy cell leukemia (HCL) is a rare type of indolent B-cell leukemia, characterized by symptoms of fatigue and weakness, organomegaly, pancytopenia, and recurrent opportunistic infections. Classic HCL should be considered a distinct clinical entity separate from HCLvariant (HCLv), which is associated with a more aggressive disease course and may not respond to standard HCL therapies. Somatic hypermutation in the IGHV gene is present in most patients with HCL. The BRAF V600E mutation has been reported in most patients with classic HCL but not in those with other B-cell leukemias or lymphomas. Therefore, it is necessary to distinguish HCLv from classic HCL. This manuscript discusses the recommendations outlined in the NCCN Guidelines for the diagnosis and management of classic HCL.

Clinical outcomes of amniotic membrane transplantation in the management of acute ocular chemical injury.

BACKGROUND: Amniotic membrane transplantation (AMT) has been used in the management of acute ocular chemical burns to promote epithelialisation, reduce inflammation and restore ocular surface integrity. The aim of this study is to analyse the morphological and functional outcomes of patients receiving AMT after ocular chemical burn. METHODS: We performed a retrospective analysis of all patients treated for acute ocular chemical burn between 1998 and 2008 in two participating centres (University of Duisburg-Essen, Germany and Royal Victoria Infirmary, Department of Ophthalmology, Newcastle University, UK). Ocular chemical burns were classified by Roper-Hall and Dua classifications. RESULTS: 72 eyes of 54 consecutive patients aged 37.3 years (±SD 11.6 years) were included in this cohort study. 7 chemical burns were acid burns, 61 were alkaline and 4 were of unknown origin. In 37 eyes (51.4%), AMT was applied within the first 6 days after injury. Mean follow-up time was 36.4 months (median 18.5; 1.3-117.3  months). Overall, 29 eyes (40.3%) achieved a best-corrected visual acuity of LogMAR 0.2 (0.63 decimal) or better at final visit. Complete 360° limbal stem cell deficiency (LSCD) occurred in 33 eyes (45.8%), while partial LSCD occurred in 21 eyes (29.2%). CONCLUSION: AMT is an effective adjunctive treatment in the management of acute ocular chemical burns to support epithelial healing and restore ocular surface integrity with potential to improve vision. However, long-term debilitated vision remained in those with severe burns complicated by LSCD.

Analysis of RegA, a pathway-specific regulator of the friulimicin biosynthesis in Actinoplanes friuliensis.

The rare actinomycete Actinoplanes friuliensis is the producer of the lipopeptide antibiotic friulimicin, which is active against a broad range of Gram-positive bacteria such as methicillin-resistant Enterococcus spec. and Staphylococcus aureus (MRE, MRSA) strains. Friulimicin consists of a decapeptide core and an acyl residue linked to an exocyclic amino acid. The complete biosynthetic gene cluster consisting of 24 open reading frames was characterized by sequence analysis and the transcription units were subsequently determined by RT-PCR experiments. In addition to several genes for biosynthesis, self-resistance and transport four different regulatory genes (regA, regB, regC and regD) were identified within the cluster. To analyse the role of the pathway-specific regulatory protein RegA in the friulimicin biosynthesis, the corresponding gene was inactivated resulting in friulimicin non-producing mutants. Furthermore, several protein-binding sites within the friulimicin gene cluster were identified by gel retardation assays. By real-time RT-PCR experiments, it was shown that the majority of the friulimicin biosynthetic genes is positively regulated by RegA.

Use of flow-cytometric analysis to optimize cell banking strategies for production of biopharmaceuticals from mammalian cells.

Production of biopharmaceuticals from mammalian cells requires generation of master, working and post-production cell banks of high quality under GMP conditions. An optimal cryopreservation strategy is needed for each new production cell line, particularly with regard to establishing production processes that are completely devoid of serum or even any animal components and to ensuring robust thaw performance for reliable production. Here, we describe a novel strategy employing flow-cytometric (FC) analysis of Annexin V-stained cells for high-throughput characterization of cell banks. Our data show that this method enables predictive evaluation of a cryopreservation strategy as early as 6h after thawing of cells. Furthermore, a broad study is presented characterizing various factors that may influence the quality of serum-free production cell banks from NSO and CHO cell lines. These results demonstrate how FC-based analysis can be used for development of future state-of-the-art cryopreservation strategies.