Laboratory diagnosis of dengue virus (DENV) infection including DENV serotyping requires skilled labor and well-equipped settings. DENV NS1 lateral flow rapid test (LFT) provides simplicity but lacks ability to identify serotype. A simple, economical, point-of-care device for serotyping is still needed. We present a gravity driven, smartphone compatible, microfluidic device using microcapillary film (MCF) to perform multiplex serotype-specific immunoassay detection of dengue virus NS1. A novel device–termed Cygnus–with a stackable design allows analysis of 1 to 12 samples in parallel in 40 minutes. A sandwich enzyme immunoassay was developed to specifically detect NS1 of all four DENV serotypes in one 60-μl plasma sample. This test aims to bridge the gap between rapid LFT and laboratory microplate ELISAs in terms of sensitivity, usability, accessibility and speed. The Cygnus NS1 assay was evaluated with retrospective undiluted plasma samples from 205 DENV infected patients alongside 50 febrile illness negative controls. Against the gold standard RT-PCR, clinical sensitivity for Cygnus was 82% in overall (with 78, 78, 80 and 76% for DENV1-4, respectively), comparable to an in-house serotyping NS1 microplate ELISA (82% vs 83%) but superior to commercial NS1-LFT (82% vs 74%). Specificity of the Cygnus device was 86%, lower than that of NS1-microplate ELISA and NS1-LFT (100% and 98%, respectively). For Cygnus positive samples, identification of DENV serotypes DENV2-4 matched those by RT-PCR by 100%, but for DENV1 capillaries false positives were seen, suggesting an improved DENV1 capture antibody is needed to increase specificity. Overall performance of Cygnus showed substantial agreement to NS1-microplate ELISA (κ = 0.68, 95%CI 0.58–0.77) and NS1-LFT (κ = 0.71, 95%CI 0.63–0.80). Although further refinement for DENV-1 NS1 detection is needed, the advantages of multiplexing and rapid processing time, this Cygnus device could deliver point-of-care NS1 antigen testing including serotyping for timely DENV diagnosis for epidemic surveillance and outbreak prediction.

Background and Aim: Feline lymphomas are categorized based on the location of tumor cells, with anatomical classifications including alimentary, mediastinal, multicentric, and extranodal forms. Accurate diagnosis and classification of feline lymphoma are paramount for enhancing treatment and prognosis. T-cell lymphomas are CD3 positive, while B-cell lymphomas exhibit positive for CD20, CD79α, and paired box 5 (PAX5). The aims of this study were (1) to classify feline lymphoma in each anatomical subtype using the World Health Organization (WHO) classification to provide information on epidemiological findings; (2) to investigate the expression and detection of B-cell lymphoma, various antibodies will be used, with the addition of PAX5, for clearer results; and (3) to gather more extensive information about feline lymphoma in Thailand, particularly in the Bangkok area. Materials and Methods: From 2011 to 2023, 86 sample tissues were submitted for routine pathological examination at the Department of Pathology, Faculty of Veterinary Science, Chulalongkorn University. Immunohistochemistry (IHC) was performed to detect an immunophenotype of PAX5, CD79α, CD20 (B-cell lineage), and CD3 (T-cell lineage). Eighty-six formalin-fixed, paraffin-embedded lymphoma tissues were prepared on silane-coated slides. After IHC, all cases were classified according to the WHO classification. Results: The most common form of lymphoma in this study was extranodal lymphoma at 37.2% (32/86), followed by multicentric lymphoma at 31.3% (27/74), mediastinal lymphoma at 17.4% (15/86), and alimentary lymphoma at 14% (12/86). Most extranodal lymphoma cases were in the nasal region. From the anatomical form, multicentric and extranodal lymphomas were predominantly diffuse large B-cell high-grade, while mediastinal lymphomas were small low-grade B-cell lymphomas. Alimentary lymphomas occur in various types, with most being the B-cell type. Conclusion: This study indicates that extranodal lymphoma and extranodal lymphoma are the most frequent presentations found in cats in Bangkok. Mediastinal and alimentary lymphomas still occur. The utilization of various B-cell markers in combination could aid pathologists in distinguishing between various stages of B-cell maturation, assessing tumor cell heterogeneity, and determining the phenotype in scenarios where there is a loss of common B-cell markers diffuse large B-cell lymphomas is the most prevalent subtype of feline lymphoma. Significantly, relying solely on immunochemistry with one parameter may not be sufficient for a definitive diagnosis of B-cell lymphoma, as another parameter may also be necessary. Keywords: B-cell, feline, immunophenotype, incidence, lymphoma, t-cell.

Background and Aim: Hematological and blood chemistry parameters are crucial for evaluating and monitoring canine multicentric lymphoma during chemotherapy. Pre-treatment hematological and blood chemistry parameters can be used as prognostic survival outcomes for this disease. Therefore, this study aimed to investigate the effect of hematological and blood chemistry parameters pre-treatment and 4 weeks post-treatment on the survival outcomes of dogs treated with either a combination of cyclophosphamide, vincristine, and prednisolone (COP) or a combination of COP with L-asparaginase (L-COP) protocols. Materials and Methods: We conducted a retrospective study. Medical records and hematological and blood chemistry parameters of 41 dogs with multicentric lymphoma treated with L-COP (n = 26) and the COP protocols (n = 15) were obtained from the hospital information system. Most cases were classified as high-grade lymphoma based on the Kiel cytological classification. The effects of hematological and blood chemistry parameters on survival outcomes were investigated using the Cox proportional hazard regression model. The median survival time (MST) for each hematological and blood chemistry parameter affecting survival outcome was established and compared using the Kaplan–Meier product limit method with the log-rank test. Results: Dogs with high-grade multicentric lymphoma that were treated with the COP protocol and had monocytosis at pre-treatment had a significantly shorter MST than dogs with normal monocyte counts (p = 0.033). In addition, dogs with azotemia, both pre-treatment and 4 weeks post-treatment, had a significantly shorter MST than dogs with normal serum creatinine levels (p = 0.012). Dogs with high-grade multicentric lymphoma treated with the L-COP protocol who had hypoalbuminemia (serum albumin concentration <2.5 mg/dL) at both pre-treatment and 4 weeks post-treatment had a significantly shorter MST than dogs with normal serum albumin levels (p < 0.001). Furthermore, dogs with leukocytosis at 4 weeks post-treatment had a significantly shorter MST than those with a normal total white blood cell count (p = 0.024). Conclusion: Serum albumin level can serve as a simple negative prognostic indicator of survival outcomes in dogs with high-grade multicentric lymphoma treated with the L-COP protocol. Dogs with hypoalbuminemia pre-treatment and 4 weeks post-treatment tended to have a shorter MST than those with normal serum albumin concentrations. Keywords: anti-cancer, chemotherapy, dogs, hypoalbuminemia, multicentric lymphoma, prognosis, retrospective study, survival outcomes.

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Background and Aim: Specific tumor biomarkers are useful for the early diagnosis of cancer or can predict the recurrence of neoplastic disease in humans and animals. Lymphoma in dogs could be classified into B-, T-, and NK-cell origins. T-cell lymphoma has the worst prognosis with a shorter survival time and disease-free interval. This study aimed to identify the differential serum protein expressions of canine B- and T-cell lymphomas compared with healthy dogs using a tandem mass tag (TMT)-based quantitative proteomics. Materials and Methods: Serum samples were collected from 20 untreated canine lymphomas (14 B-cells and 6 T-cells) and four healthy control dogs. Sera peptides from each sample were processed for TMT 10-plex tagging and analyzed using liquid chromatography-mass spectrometry (MS). Differential proteome profiling was then compared between lymphoma and control. Results: We discovered 20 elevated and 14 decreased serum proteins in the lymphoma group relative to the healthy group. Six candidate increased proteins in canine lymphomas were beta-actin cytoplasmic 1 (ACTB, p=0.04), haptoglobin (p=0.002), beta-2 microglobulin (β2M, p=0.007), beta-2 glycoprotein 1 (APOH, p=0.03), metalloproteinase inhibitor 1 (TIMP-1, p=0.03), and CD44 antigen (p=0.02). When compared between B- and T-cell lymphomas, B-cell phenotypes had upregulated immunoglobulin (Ig) heavy chain V region GOM (p=0.02), clusterin (p=0.01), apolipoprotein C1 (APOC1, p=0.05), and plasminogen (p=0.02). Conclusion: These findings were investigated quantitative serum proteomes between B- and T-cell lymphomas using TMT-based MS. ACTB, β2M, APOH, TIMP-1, CD44 antigen, Ig heavy chain V region GOM, and APOC1 are novel candidate proteins and might serve as a lymphoma biomarker in dogs. However, evaluation with an increased sample size is needed to confirm their diagnostic and prognostic ability.