A Computational Framework for Comprehensive Genomic Profiling in Solid Cancers: The Analytical Performance of a High-Throughput Assay for Small and Copy Number Variants.

In January 2022, our institution launched a comprehensive cancer genome profiling program on 10 cancer types using a non-IVD solution named the TruSight Oncology 500 Assay provided by Illumina®. The assay analyzes both DNA and RNA, identifying Single-Nucleotide Variants (SNV)s and Insertion-Deletion (InDel) in 523 genes, as well as known and unknown fusions and splicing variants in 55 genes and Copy Number Alterations (CNVs), Mutational Tumor Burden (MTB) and Microsatellite Instability (MSI). According to the current European IVD Directive 98/79/EC, an internal validation was performed before running the test. A dedicated open-source bioinformatics pipeline was developed for data postprocessing, panel assessment and embedding in high-performance computing framework using the container technology to ensure scalability and reproducibility. Our protocols, applied to 71 DNA and 64 RNA samples, showed full agreement between the TruSight Oncology 500 assay and standard approaches, with only minor limitations, allowing to routinely perform our protocol in patient screening.

Integrating a Comprehensive Cancer Genome Profiling into Clinical Practice: A Blueprint in an Italian Referral Center.

The implementation of cancer molecular characterization in clinical practice has improved prognostic re-definition, extending the eligibility to a continuously increasing number of targeted treatments. Broad molecular profiling technologies better than organ-based approaches are believed to serve such dynamic purposes. We here present the workflow our institution adopted to run a comprehensive cancer genome profiling in clinical practice. This article describes the workflow designed to make a comprehensive cancer genome profiling program feasible and sustainable in a large-volume referral hospital.

Solvent influence on manufacturability, phase behavior and morphology of amorphous solid dispersions prepared via bead coating.

Bead coating or fluid-bed coating serves as an auspicious solvent-based amorphous solid dispersion (ASD) manufacturing technique in respect of minimization of potential physical stability issues. However, the impact of solvent selection on the bead coating process and its resulting pellet formulation is, to the best of our knowledge, never investigated before. This study therefore aims to investigate the influence of the solvent on the bead coating process itself (i.e. manufacturability) and on solid-state characteristics of the resulting ASDs coated onto beads. For this purpose, the drug-polymer system felodipine (FEL)-poly(vinylpyrrolidone-co-vinyl acetate) (PVP-VA) was coated onto microcrystalline cellulose (MCC) beads from acetonitrile (ACN), methanol (MeOH), ethanol (EtOH), acetone (Ac), 2-propanol (PrOH), dichloromethane (DCM) and ethyl acetate (EthAc). A drug loading screening approach with bead coating revealed analogous ability to manufacture high drug-loaded ASDs from the different organic solvents. The results show no correlation with crystallization tendency or with equilibrium solubility of the drug in the different solvents, nor with the solvent-dependent drug-polymer miscibility obtained from film casting experiments. Distinct coating morphologies were however observed for PVP-VA and FEL-PVP-VA ASDs deposited onto beads from the various solvents, which is attributed to differences in solvent evaporation kinetics.

Estafilococcia em paciente com tricoleucemia: relato de caso / Staphylococci in patient with hairy cell leukemia. Case report

JUSTIFICATIVA E OBJETIVOS: A leucemia de células pilosas (LCP) é caracterizada clinicamente por esplenomegalia, pancitopenia e raramente acomete linfonodos. Nos casos mais graves de leucopenia a predisposição às infecções é mais acentuada. Apesar dos métodos diagnósticos para a LCP estarem bem estabelecidos, o objetivo deste estudo foi alertar para o fato de que infecções persistentes podem indicar imunossupressão, devendo portanto, lembrar da LCP como hipótese diagnóstica. RELATO DO CASO: Paciente do sexo masculino, 47 anos, deu entrada no pronto-socorro com quadro clínico de estafilococcia. A cultura do material da punção mostrou a presença de Staphylococcusaureus. Foi realizado mielograma, cujo aspirado mostrou hipocelularidade global, a biópsia concluiu aplasia de medula ósseaque em associação com a imuno-histoquímica mostrou umprocesso linfoproliferativo de células B, que sugeriu continuar ainvestigação. Posteriormente foi realizado a imunofenotipagemem painel proliferativo de sangue periférico com perfil positivopara: CD20**, cadeia leve lambda*, FMC-7, CD19, CD79b,IgM, IgD, CD11c**, CD25, CD200, CD22, CD103, CD123,CD45 e bcl-2, compatível com LCP.CONCLUSÃO: Pacientes com neoplasias hematológicas são acometidos frequentemente por infecções bacterianas na corrente sanguínea, sendo o Staphylococcus aureus amplamente isoladonas hemoculturas.

BACKGROUND AND OBJECTIVES: Hairy cell leukemia(HCL) is characterized clinically by splenomegaly, pancytopenia and the lymphonodes are rarely palpable. In more sever cases of leukopenia, predisposition to infections is more pronounced. Although the diagnostic methods for the HCL are well established, the objective of this study was to draw attention to the fact that persistent infections may indicate immunosuppression, and therefore remember the HCL as a diagnostic hypothesis. CASE REPORT: Male patient, 47 year-old, was admitted to the hospital with clinical staphylococci. Material culture punctures howed the presence of Staphylococcus aureus. Myelogram was performed, which showed global hypocellular aspirate, the bone marrow biopsy concluded for bone marrow aplasia that inassociation with immunohistochemistry showed a lymphoproliferative process of B cell, which suggested keep the investigation. After that was performed a immunophenotyping panel of prolife rative peripheral blood that showed a positive profile for: CD20**, lambda light chain**, FMC7, CD19, CD79b, IgM,IgD, D11c **, CD25, CD200, CD22, CD103, CD123, CD45 and bcl2, consistent with HCL. CONCLUSION: Patients with hematological malignancies are often affected by bacterial infections in the bloodstream, where Staphylococcus aureus was largely isolated in blood cultures.