Evaluation of high-fluorescence body fluid (HF-BF) parameter as a screening tool of malignancy in body fluids

Introduction: Automated body fluid (BF) analysis is gradually replacing the traditional methods of cell counting in all BFs. This study was done to analyze the high-fluorescence (HF)-BF parameter generated on Sysmex XN-1000 and study its correlation with the presence of malignant cells in the body fluids. A correlation between manual and automated differential counts was also done. Materials and Methods: A total of 1985 samples including 797 ascitic fluids (AF), 532 pleural fluids (PF), and 656 cerebrospinal fluids (CSF) were run on Sysmex XN-1000 in BF mode and cytopathology was available for 924 BFs including 389 AF, 379 PF, and 156 CSF. Both manual and automated methods were used for cell differential and cell morphology. Results: Of the 924 samples with corresponding cytopathology, malignancy was found in 59 samples. The HF-BF%/100 WBCs (24.8 ± 72.5) and HF-BF#/?L (329.86 ± 932.35) for malignant BF samples were found to be significantly higher than the nonmalignant samples (4.41 ± 8.1) and (19.57 ± 61.91), respectively. Receiver–operator-characteristic curve cutoffs for all BF for percentage and absolute HF-BF were 2.85%/100 WBCs and >12/?L. A good correlation was found between the manual and automated WBC differential counts in all fluids except CSF with total count <5/?L. Conclusions: BFs can be reliably analyzed on automated analyzers. HF-BF parameter is helpful in identifying malignant samples but cannot be totally relied upon. If HF-BF%/# are above the lab-generated cutoffs, microscopy should be done. A complete validation study on HF-BF parameter in BF mode is desired to set the standards for the analysis of serious effusions.

Platelet function tests: A 5-year audit of platelet function tests done for bleeding disorders in a tertiary care center of a developing country

Introduction: The platelet function disorders remain largely undiagnosed or incompletely diagnosed in developing nations due to lack of availability of tests like lumiaggregometry, granule release assay or molecular testing. We performed a retrospective analysis of all the platelet function test (PFT) carried out in past 5 years by Light transmission aggregometery (LTA) using a panel of agonist. The indications and the test results were analyzed by two hematopathologist with the aim to look into the present diagnostic facilities or lack of it for correct diagnosis. This is essential for better management and genetic counselling. Materials and Methods: The PFT was performed both on patients and healthy unrelated age specific controls by light transmission aggregometry on Chronolog platelet aggregometer using platelet rich plasma. The panel of agonists included ADP (10?m/l and 2.0 ?m/l), epinephrine (10.0 ?m/l), collagen (2?g/ml), arachidonic acid (0.75 mM) and ristocetin (1.25 mg/ml & 0.25 mg/l). Results: The 5 years records of 110 cases were audited, 101 of these were tested for clinical bleeding , 35 adults and 66 children. The adults included 29 women and 6 men, 17 to 82 years of age. The children were 16 years to 3 months of age, 30 girls and 36 boys. Platelet function test abnormality was found in 31.6% (32/101) cases ,a majority remained undiagnosed of these about 21% had clinically significant bleeding.The cases diagnosed included Glanzmann Thromboasthenia-11 , von Willebrand Disease-6, Bernard Soulier'syndrome-1, storage pool disorder-6, mild defect of Epinephrine-3, isolated defect with collagen in1. Conclusion: An epidemiologically large proportion of platelet function disorders amongst people living in developing nations remain undiagnosed. This lacunae needs to be highlighted and addressed on larger scale. The options available are to increase the available armamentarium of tests or international collaboration with a specialized laboratory to aid in complete diagnosis.

Computational studies on PLK1 delta gene in cancer diseases.

Cancer is a group of diseases involving abnormal cell growth with the potential to invade or spread to other parts of the body. Polo-like kinases (Plks) are a family of conserved serine/threonine kinases involved in the regulation of cell cycle progression through G2 and mitosis. One of them being PLK1, it’s over expression leads to a variety of cancers. Plk1 delta, an uncharacterized protein sequence found in UniProt was studied and found to consist of the N-terminal portion of plk1gene. The 3D protein structure of PLK1 delta was modeled and then the predicted structure was validated. Molecular dynamics simulation was performed to find the stability of the protein. The modelled protein was docked to BI 2536, an inhibitor of Plk1 to obtain conformation of least binding energy. The interacting sites between the protein and inhibitor were also analyzed.

Exploring novel drug targets in fatty acid pathway of Plasmodium falciparum.

The malarial parasite Plasmodium falciparum infects humans and proliferates rapidly inside the host before its detection. The proliferation step requires a large amount of lipids for membrane synthesis. Thus fatty acid biosynthesis occurring in the apicoplast plays an important role in causing cerebral malaria. In this study, we explored and analyzed these pathways using stoichiometric matrix, elementary flux modes and robustness analysis. Based on the above analysis, the robustness of this pathway diminished as the result of virtual enzyme knock out indicating four key enzymes, 3-oxoacyl-ACP synthase, 3-oxoacyl-ACP synthase, 3-oxoacyl-ACP synthase and Glycerol-3-phosphate o-acyl transferase. Among the four, the first three are existing drug targets. Subsequently, we also found that a combinatorial double knock out of these enzymes predicts further reduction in overall pathway enzyme activity. Thus, we propose multi drug targeting as a better way to treat brain malaria.

Clinico‑hematological profile and outcome of acute promyelocytic leukemia patients at a tertiary care center in North India.

OBJECTIVES: Acute promyelocytic leukemia (APL) is the only acute leukemia amenable to targeted therapy. However, there is limited Indian data on APL. We aimed to analyze data of APL patients treated with all trans retinoic acid (ATRA) and anthracycline based chemotherapy. MATERIALS AND METHODS: A total of 34 cases of APL were treated at our center over 4 years. Induction chemotherapy consisted of a combination of ATRA and daunorubicin. RESULTS: Most of our patients were intermediate risk (50%) followed by high risk (41.17%). Induction mortality was 14.7%. We observed a high incidence of febrile neutropenia (91%) and 50% of our patients developed ATRA syndrome. Four patients (11.76%) relapsed during follow‑up (median ‑ 15 months, range: 13‑33 months). There was no correlation between risk status and death or relapse or ATRA syndrome. Median event free survival (EFS) duration was not reached however mean duration was 38.45 ± 3.84 months. Median overall survival (OS) duration was also not reached at 53 months of follow‑up. The 4 year OS and EFS were 75.45% and 64.5% respectively. On multivariate analysis, only disseminated intravascular coagulation (DIC) significantly correlated with a poor OS and EFS. DISCUSSION: Our data reflects that APL remains a highly curable malignancy with good response to ATRA plus anthracycline based chemotherapy even with a greater number of high and intermediate risk patients. Only DIC during induction chemotherapy bore an impact on survival in our patients.