Development, characterization, and evaluation of withaferin-A and artesunate-loaded pH-responsive acetal-dextran polymeric nanoparticles for the management of malaria.

Artemisinin and its derivatives have been commonly used to treat malaria. However, the emergence of resistance against artemisinin derivatives has posed a critical challenge in malaria management. In the present study, we have proposed a combinatorial approach, utilizing pH-responsive acetal-dextran nanoparticles (Ac-Dex NPs) as carriers for the delivery of withaferin-A (WS-3) and artesunate (Art) to improve treatment efficacy of malaria. The optimized WS-3 and Art Ac-Dex NPs demonstrated enhanced pH-responsive release profiles under parasitophorous mimetic conditions (pH 5.5). Computational molecular modeling reveals that Ac-Dex's polymeric backbone strongly interacts with merozoite surface protein-1 (MSP-1), preventing erythrocyte invasion. In-vitro antimalarial activity of drug-loaded Ac-Dex NPs reveals a 1-1.5-fold reduction in IC50 values compared to pure drug against the 3D7 strain of Plasmodium falciparum. Treatment with WS-3 Ac-Dex NPs (100 mg/kg) and Art Ac-Dex NPs (30 mg/kg) to Plasmodium berghei-infected mice resulted in 78.11 % and 100 % inhibition of parasitemia. Notably, the combination therapy comprised of Art and WS-3 Ac-Dex NPs achieved complete inhibition of parasitemia even at a half dose of Art, indicating the synergistic potential of the combinations. However, further investigations are necessary to confirm the safety and effectiveness of WS-3 and Art Ac-Dex NPs for their successful clinical implications.

A Comparative Evaluation of Performance of Sysmex XN 3000 and Horiba Yumizen H2500 Automated Complete Blood Count Analysers.

Modern automated laboratory haematology analysers use various methods to measure different haematological parameters. These parameters are useful in the diagnostic and clinical interpretation of patient symptoms. So, it is very important to compare the performance of different analysers measuring the same parameter. Hence, a comparison of complete blood counts analysed by Sysmex XN 3000 and Horiba Yumizen H2500 was performed. Total 296 EDTA anti-coagulated blood samples were processed in both the analysers in duplicate within 4 h of collection. The white blood cell count, red blood cell count, erythrocyte indices, differential leukocyte count, platelet count and platelet indices and reticulocyte count were compared. A good level of correlation and agreement between different parameters were obtained. A strong correlation was observed (r > 0.9) between Sysmex XN 3000 and Yumizen H2500 for WBC (0.997), RBC (0.997), Haemoglobin (0.999), haematocrit (0.974), MCV (0.902), MCH (0.99),, platelet count by impedance (0.989), mean platelet volume (0.954), plateletcrit (0.971), platelet distribution width (PDW) (0.916), neutrophils (0.997), lymphocytes (0.989), monocytes (0.943), and eosinophils (0.991) counts. A moderate correlation was observed for RDW-CV (0.75). The basophils count showed poor correlation (r < 0.5) possibly because of sample selection with mostly low basophils count. An acceptable bias was observed for most of the parameters like WBC, RBC, Haemoglobin, Haematocrit, platelet counts, neutrophils, lymphocytes, eosinophils and monocytes. The studied instruments ensured satisfactory interchangeability except for few parameters, thus facilitate substitution of one analyser by another without affecting the clinical decision making.

Identification of starvation-mimetic bioactive phytocomponent from Withania somnifera using in-silico molecular modelling and flow cytometry-based analysis for the management of malaria.

Multidrug resistance episodes in malaria increased from 3.9% to 20% from 2015 to 2019. Synchronizing the clinical manifestation in chronological sequence led to a unique impression on glucose demand (increased up to 100-fold) by the parasite-infected RBCs. Hence, restriction in the glucose uptake to parasite-infected RBCs could be an alternative approach to conquer the global burden of malaria to a greater extent. A C28 steroidal lactone Withaferin A (WS-3) isolated from Withania somnifera leave extract shows better thermodynamically stable interactions with the glucose transporters (GLUT-1 and PfHT) to standard drugs metformin and lopinavir. MD simulations for a trajectory period of 100 ns reflect stable interactions with the interactive amino acid residues such as Pro141, Gln161, Gln282, Gln283, Trp388, Phe389, and Phe40, Asn48, Phe85, His168, Gln169, Asn311 which potentiating inhibitory activity of WS-3 against GLUT-1 and PfHT respectively. WS-3 was non-hemotoxic (%hemolysis <5%) for a high concentration of up to 1 mg/ml in the physiological milieu. However, the %hemolysis significantly increased up to 30.55 ± 0.929% in a parasitophorous simulated environment (pH 5.0). Increased hemolysis of WS-3 could be due to the production of ROS in an acidic environment. Further, the inhibitory activity of WS-3 against both glucose transporters was supported with flow cytometry-based analysis of parasite-infected RBCs. Results show that WS-3 has low mean fluorescence intensities for both target proteins compared to conventional drugs, suggesting a potential sugar transporter inhibitor against GLUT-1 and PfHT for managing malaria. Communicated by Ramaswamy H. Sarma.

Understanding the functional relevance of oral neutrophils, phenotype and properties in OSCC.

Neutrophils are the predominant white blood cells (WBC) that are recruited to the sites of inflammation and infection. They are acknowledged to perform dual roles by promoting (pro-tumor) or by exhibiting anti-cancer properties (anti-tumor). Neutrophils are characterized based on the changes in phenotype and functional properties. To this context, circulating polymorphonuclear neutrophils (cPMN) and tumor-associated neutrophils (TANs) in cancer biology has been well explored but limited to oral polymorphonuclear neutrophils (oPMNs) in oral squamous cell carcinoma (OSCC). However, oPMNs are eminent in maintaining the healthy oral ecosystem by neutralizing microorganisms. Neutralization process enhances the expression of cell surface markers (CD11b, CD63, CD66, CD66b, CD66c, and CD66e) and inflammatory cytokines (TNF-α, IFN-γ, GM-CSF, and IL-8) and increases the recruitment of neutrophils. Along with the inflammation, it has been reported that CEACAM1 and chemerin also favors the infiltration of neutrophils to the cancer site. This indicates that oPMN might contribute to the aetiology of OSCC. The main objective of this review is to explore, the production and migration of oPMNs to the oral cavity, their phenotypes and possible role in OSCC.

Phytochemical investigation, structural elucidation, in silico study and anti-psoriatic activity of potent bioactive from Betula utilis.

Psoriasis is a chronic autoimmune pathological condition characterized by hyperactivation of proinflammatory cytokines (IL-6, TNF-α, IL-17, IL-23, etc.). Severe drug-associated toxicities like hepatotoxicity and nephrotoxicity (Methotrexate), teratogenicity (Tazarotene), hypercholesterolemia (Cyclosporine) and hypercalcemia (tacalcitol), are the forefront challenges that demand an alternative approach for the treatment of psoriasis. In the present study, a natural lead molecule 'Betulin' (BE, lup-20(29)-ene-3b,28-diol) was isolated from Betula utilis and subsequently, structure-based molecular docking was employed to identify the molecular target for psoriasis. The computational analysis reflects better affinity of BE towards pro-inflammatory cytokine as compared to standard drugs. Apart from this BE shows a greater affinity towards the overexpressed Glut-1 receptor in comparison to standard drug Metformin (Met). Based on the in silico screening the isolated lead compound was further processed for the evaluation of anti-psoriatic activity via imiquimod (IMQ 5%) induced psoriasis-like skin inflammation model. In vivo screening models were characterized by different parameters (psoriasis area and severity index (PASI) scores, macroscopically and behavioral evaluation, splenomegaly, cytokine levels and histological changes) and compared among the experimental groups. The experimental finding reflects comparable results of PASI score, i.e., 57.14% and 61.9% recovery of test BE-solution (180 mg/kg) and standard Betamethasone di-propionate ointment (BD-oint.0.5 mg/g), respectively. Focusing on other parameters, BE shows relative results such as an enhanced macroscopically with behavioral conditions, reducing the expression of proinflammatory cytokine as well as restoring histological changes with that of BD. These findings suggest that BE-isolated phytoconstituents from Betula utilis could be a potential agent and a step closer to psoriasis treatment. HIGHLIGHTPsoriasis is a multifaceted, immunologically mediated disease consequences production of high levels of proinflammatory mediators and overexpression of Glut-1 transporters that trigger keratinocyte proliferation and inflammatory cascades.A Himalayan silver birch, Betula utilis (Bhojpatra) contains many steroidal terpenes which are responsible for various pharmacological activities that could be exploited in drug development in psoriasis.The computational analysis of BE reflects a better affinity toward the proinflammatory cytokines with their target receptors and indicates a satisfactory range with a slight deviation from Jorgensen and Lipinski's rule and possesses a significant drug choice for psoriasis.Preclinical findings of BE-solution (BE-sol) give a positive response towards IMQ-induced psoriasis-like skin inflammation model.[Figure: see text]Communicated by Ramaswamy H. Sarma.

An event of Evans even in HIV.

Autoimmune cytopenias may be the initial presentation in patients with HIV infection or can develop while on treatment with antiretroviral therapy (ART). These cytopenias usually resolve after initiation of ART. We report a rare case of HIV who presented with Evans syndrome on ART, being refractory to steroids and rituximab but with response to splenectomy.

Challenges in Detection of Adolescent Anaemia: Validation of Point-of-Care Device (Mission® plus) for Haemoglobin Measurement among Tribal Residential School Children of Selected Districts of Odisha, India.

BACKGROUND: Screening for anemia among tribal school children has been a challenge. OBJECTIVES: To validate a point-of-care (POC) device (mission® plus hemoglobinometer) to the gold standard method, spectrophotometry. STUDY DESIGN: Cross-sectional study. PARTICIPANTS: The representative sample of 953 tribal adolescents from the residential schools of Odisha. METHODS: Hemoglobin was measured simultaneously by the POC and gold standard method during January to July 2019. The validity of the POC device was measured by sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). The area under the curve was calculated using receiver operating characteristic (ROC) analysis. Concordance of the POC device with the gold standard method was determined by the Bland-Altman plot. The intraclass correlation coefficient (ICC), precision (⍴), a bias correction factor (Cb), and the concordance correlation coefficient were also calculated. Deming regression analysis was performed, and a linear equation was established. RESULTS: The mean age of the study participants was 13.07 (±1.48) years. The prevalence of anemia was 45.54% by the gold standard method. The sensitivity and specificity of the POC device were 94.9% and 56.1%, respectively. PPV and NPVs were 64.4% and 93.0%, respectively. The area under the ROC curve was found to be 0.856. The ICC was 0.887 (95% confidence interval: 0.872-0.901). CONCLUSIONS: Very good reliability/absolute agreement for hemoglobin measurements existed between the POC device and the gold standard method making it suitable as a screening device.

Flow Cytometry Based MRD and Its Impact on Survival Outcome in Children and Young Adults with ALL: A Prospective Study from a Tertiary Cancer Centre in Southern India.

Presence of minimal residual disease (MRD) following induction chemotherapy is a well-recognized risk factor to predict relapse in acute lymphoblastic leukemia (ALL). There is paucity of data on MRD and outcome in ALL from India. We share our experience in establishing a flow cytometry-based MRD assay for ALL with emphasis on determination of the number of patients who had MRD on day 35 of induction therapy and its correlation with outcome and other prognostic factors. We prospectively studied MRD in patients with ALL less than 25 years who achieved morphological complete remission with induction therapy. The initial series consisted of 104 patients with ALL. Ninety-two patients had bone marrow samples collected on day 35 of remission induction chemotherapy that was adequate for MRD. Strategy of monitoring MRD was based on flow cytometry using six color staining according the leukemia associated immunophenotype found at diagnosis. Data analysis was done using Fisher exact test. The median age was 8.5 years (range 0.9-22 years). Thirty-seven out of ninety-two patients (40.2%) had MRD at end of induction. MRD on day 35 was between 0.01 and 0.1% in 18.9% of patients, between 0.1 and 1% in 59.5% and more than 1% in 21.6% patients. Among the patients who had MRD, 16.7% had favourable cytogenetics, 60% had intermediate and 13.3% had high-risk cytogenetics. The presence or absence of residual leukemia by flow cytometry at day 35 was not significantly related to age (p = 1.0), male gender (p = 0.08) hyperleukocytosis (p = 0.25) or day 8 blast clearance (p = 0.21). However, T cell phenotype (p < 0.001) was significantly associated with MRD. The 5-year event free survival (EFS) for patients who had MRD versus those who did not was 69% and 61.1% respectively (p = 0.41). The 5-year overall survival (OS) for patients who had MRD versus those who did not was 72.5% and 61.1% respectively (p = 0.33). Flow cytometric techniques can be applied to monitor MRD in patients of ALL undergoing induction therapy. Our results suggest MRD correlates with certain known prognostic factors. Though the EFS and OS was lower in MRD positive patients, the results were not statistically significant probably because of the small sample size.