Ultracellular Imaging of Bronchoalveolar Lavage from Young COVID-19 Patients with Comorbidities Showed Greater SARS-COV-2 Infection but Lesser Ultrastructural Damage Than the Older Patients.

In this study, we examined the cellular infectivity and ultrastructural changes due to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in the various cells of bronchoalveolar fluid (BALF) from intubated patients of different age groups (≥60 years and <60 years) and with common comorbidities such as diabetes, liver and kidney diseases, and malignancies. BALF of 79 patients (38 cases >60 and 41 cases <60 years) were studied by light microscopy, immunofluorescence, scanning, and transmission electron microscopy to evaluate the ultrastructural changes in the ciliated epithelium, type II pneumocytes, macrophages, neutrophils, eosinophils, lymphocytes, and anucleated granulocytes. This study demonstrated relatively a greater infection and better preservation of subcellular structures in these cells from BALF of younger patients (<60 years compared with the older patients (≥60 years). The different cells of BALF from the patients without comorbidities showed higher viral load compared with the patients with comorbidities. Diabetic patients showed maximum ultrastructural damage in BALF cells in the comorbid group. This study highlights the comparative effect of SARS-CoV-2 infection on the different airway and inflammatory cells of BALF at the subcellular levels among older and younger patients and in patients with comorbid conditions.

One-Pot Crystallization of 2D and 3D Cobalt-Based Metal-Organic Frameworks and Their High-Performance Electrocatalytic Oxygen Evolution.

Developing highly competent and low-cost earth-plentiful metal-based oxygen evolution reaction (OER) electrocatalysts is critical for future electrochemical conversion technologies and renewable energy systems. Herein, two cobalt-based metal-organic frameworks (Co-MOFs), [Co6(btc)2(DMF)6(HCOO)6] (2D, MOF 1) and [Co3(CHOO)9{DMA}3] (3D, MOF 2), where btc = 1,3,5-benzenetricarboxylic acid, DMF = N,N-dimethylformamide, and DMA = dimethylamine, have been crystallized under hydrothermal conditions, from a single reaction. MOF 1 shows an extraordinary OER performance with 175 mV overpotential to attain 10 mA cm-2 current density with a low Tafel slope value of 80 mV dec-1, whereas MOF 2 achieves 10 mA cm-2 current density at 389 mV overpotential. Two different architecture-based MOFs have been synthesized from a single solution for the very first time. Also, the OER activity of MOF 1 overpowers the commercially used RuO2 and surpasses most of the reported OER electrocatalysts. Post OER characterization of MOF 1 revealed the in situ formation of Co(OH)2 and CoOOH, acting as active sites for the OER process.

A polyoxomolybdate-based hybrid nano capsule as an antineoplastic agent.

Polyoxometalates (POMs) are versatile anionic clusters which have attracted a lot of attention in biomedical investigations. To counteract the increasing resistance effect of cancer cells and the high toxicity of chemotherapeutic treatments, POM-based metallodrugs can be strategically synthesized by adjusting the stereochemical and physicochemical features of POMs. In the present report a polyoxomolybdate (POMo) based organic-inorganic hybrid solid (C6H16N)(C6H15N)2[Mo8O26]·3H2O, solid 1, has been synthesized and its antitumoral activities have been investigated against three cancer cell lines namely, A549 (Lung cancer), HepG2 (Liver cancer), and MCF-7 (Breast cancer) with IC50 values 56.2 µmol L-1, 57.3 µmol L-1, and 55.2 µmol L-1 respectively. The structural characterization revealed that solid 1 consists of an octa molybdate-type cluster connected by three triethylamine molecules via hydrogen bonding interactions. The electron microscopy analysis suggests the nanocapsule-like morphology of solid 1 in the size range of 50-70 nm. The UV-vis absorption spectra were used to assess the binding ability of synthesized POM-based solid 1 to calf thymus DNA (ctDNA), which further explained the binding interaction between POMo and ctDNA and the binding constant was calculated to be 2.246 × 103 giving evidence of groove binding.

Impact of prophylactic hydroxychloroquine on ultrastructural impairment and cellular SARS-CoV-2 infection in different cells of bronchoalveolar lavage fluids of COVID-19 patients.

Many drugs were recommended as antiviral agents for infection control and effective therapy to reduce the mortality rate for COVID-19 patients. Hydroxychloroquine (HCQ), an antimalarial drug, has been controversially recommended for prophylactic use in many countries, including India, to control SARS-CoV-2 infections. We have explored the effect of prophylactic HCQ from the cells of bronchoalveolar lavage fluids from COVID-19-induced acute respiratory distress syndrome patients to determine the level of infection and ultrastructural alterations in the ciliated epithelium, type II pneumocytes, alveolar macrophages, neutrophils, and enucleated granulocytes. Ultrastructural investigation of ciliated epithelium and type II pneumocytes showed lesser infections and cellular impairment in the prophylactic HCQ+ group than HCQ- group. However, macrophages and neutrophils displayed similar infection and ultrastructural alterations in both patient groups. The enucleated fragments of granulocytes showed phagocytosis of the matured virus in HCQ+ groups. The present report unveils the ultrastructural proof to complement the paradox regarding the role of prophylactic HCQ in COVID-19 patients.

A rare polyoxometalate cluster [NiW12O44]14- based solid as a pre-catalyst for efficient and long-term oxygen evolution.

Polyoxometalates (POMs) are an eminent class of metal oxide anionic clusters of early transition metals with huge structural diversity. Herein, a [NiW12O44]14- cluster based solid, (C5H7N2)6[NiW12O44], has been reported (PS-78). The [NiW12O44]14- cluster bridges the missing gap of 1 : 12 hetero-POMs of Keggin and Silverton together with a coordination number of 8 of the central heteroatom (Ni). Furthermore PS-78 has been explored as an efficient and highly sustained oxygen evolution pre-catalyst in alkaline medium with an overpotential of 347 mV to attain a current density of 10 mA cm-2 and long-term stability up to 96 hours. Furthermore, mechanistic investigation showed that in situ generated NiO and WO x (x = 1, 2) species act as active species for the oxygen evolution reaction. This study will open up new avenues for exploring POMs' new topologies and the potential of POMs as effective pre-catalysts in electrocatalytic applications.

An inhibitor of RORγ for chronic pulmonary obstructive disease treatment.

The role of RORγ as a transcription factor for Th17 cell differentiation and thereby regulation of IL-17 levels is well known. Increased RORγ expression along with IL-17A levels was observed in animal models, immune cells and BAL fluid of COPD patients. Increased IL-17A levels in severe COPD patients are positively correlated with decreased lung functions and increased severity symptoms and emphysema, supporting an urgency to develop novel therapies modulating IL-17 or RORγ for COPD treatment. We identified a potent RORγ inhibitor, PCCR-1 using hit to lead identification followed by extensive lead optimization by structure-activity relationship. PCCR-1 resulted in RORγ inhibition with a high degree of specificity in a biochemical assay, with > 300-fold selectivity over other isoforms of ROR. Our data suggest promising potency for IL-17A inhibition in human and canine PBMCs and mouse splenocytes with no significant impact on Th1 and Th2 cytokines. In vivo, PCCR-1 exhibited significant efficacy in the acute CS model with dose-dependent inhibition of the PD biomarkers that correlated well with the drug concentration in lung and BAL fluid, demonstrating an acceptable safety profile. This inhibitor effectively inhibited IL-17A release in whole blood and BALf samples from COPD patients. Overall, we identified a selective inhibitor of RORγ to pursue further development of novel scaffolds for COPD treatment.

A Mo(VI) based coordination polymer as an antiproliferative agent against cancer cells.

Metal ions being an important part of biological systems are of great interest in the designing of new drugs. Molybdenum is an essential trace element for humans, animals, and plants and naturally present in many enzymes hence its complexes can be expected to serve as potential candidates for biomedical applications. A novel molybdenum-based coordination polymer, [Mo2(µ2-O)O4(2-pyc)2(H2O)], is synthesized by a hydrothermal route and structurally characterized by using single crystal X-Ray diffraction. The structure consists of molybdenum octahedra connected by a bridging oxo ligand and 2-pyc forming a one-dimensional coordination polymer. This Mo coordination polymer was found to show a considerable inhibitory effect with IC50 values of 22.63 µmol L-1, 28.19 µmol L-1, and 20.97 µmol L-1, against HepG2 (human liver cancer), A549 (human lung cancer), and MCF-7 (human breast cancer) cell lines respectively. This is the first attempt at exploring the molybdenum-based coordination polymer for antitumor applications. The cell cytotoxicity analysis revealed that the anti-tumor potential of the compound is governed by arresting of the A549, HepG2, and MCF-7 cancer cells in the S phase of the cell cycle. UV-Visible absorption spectroscopy further revealed the binding interaction between the Mo coordination polymer and ctDNA and the binding constant was found to be 5.9 × 103 L mol-1, which is in agreement with those of well-known groove binders. This binding interaction in turn induces apoptosis and necrosis pathways leading to the death of the cancer cells.

Prenatal Diagnosis by Chromosome Microarray Analysis, An Indian Experience.

BACKGROUND: Karyotyping has been the gold standard for prenatal chromosome analysis. The resolution should be higher by chromosome microarray analysis (CMA). The challenge lies in recognizing benign and pathogenic or clinically significant copy number variations (pCNV) and variations of unknown significance (VOUS). The aim was to evaluate the diagnostic yield and clinical utility of CMA, to stratify the CMA results in various prenatal referral groups and to accumulate Indian data of pCNVs and VOUS for further interpretation to assist defined genetic counseling. METHODS: Karyotyping and CMA were performed on consecutive referrals of 370 prenatal samples of amniotic fluid (n = 274) and chorionic villi (n = 96) from Indian pregnant women with high maternal age (n = 23), biochemical screen positive (n = 61), previous child abnormal (n = 59), abnormal fetal ultrasound (n = 205) and heterozygous parents (n = 22). RESULTS AND CONCLUSION: The overall diagnostic yield of abnormal results was 5.40% by karyotyping and 9.18% by CMA. The highest percentage of pCNVs were found in the group with abnormal fetal ultrasound (5.40%) as compared to other groups, such as women with high maternal age (0.81%), biochemical screen positive (0.54%), previous abnormal offspring (0.81%) or heterozygous parents group (1.62%). Therefore, all women with abnormal fetal ultrasound must undergo CMA test for genotype-phenotype correlation. CMA detects known and rare deletion/duplication syndromes and characterizes marker chromosomes. Accumulation of CNV data will form an Indian Repository and also help to resolve the uncertainty of VOUS. Pretest and posttest genetic counseling is essential to convey benefits and limitations of CMA and help the patients to take informed decisions.

Effective inhibitory activity against MCF-7, A549 and HepG2 cancer cells by a phosphomolybdate based hybrid solid.

A novel Strandberg type polyoxomolybdate based organic-inorganic hybrid solid, [{4,4'-H2bpy}{4,4'-Hbpy}2{H2P2Mo5O23}]·5H2O (1) has been synthesized and structurally characterized by the single crystal X-ray diffraction technique. The structure consists of a discrete type phosphomolybdate cluster, [H2P2Mo5O23]4-, connected with three protonated 4,4'-bipyridine molecules by strong hydrogen bonding interactions. The In vitro anti-tumoral activity of compound (1) was tested against human breast cancer (MCF-7), human lung cancer (A549) and human liver cancer (HepG2) cells. The Strandberg type cluster was used against the MCF-7 and A549 cancer cells for the first time hitherto. It shows considerable inhibitory effect with IC50 values of 33.79 µmol L-1, 25.17 µmol L-1, and 32.11 µmol L-1 against HepG2, A549 and MCF-7 respectively. The anti-tumoral activity of 1 was also found to be comparable with that of a routinely used chemotherapeutic agent, methotrexate (MTX), with an IC50 value of 42.03 µmol L-1 for HepG2, 26.93 µmol L-1 for A549 and 49.79 µmol L-1 for MCF-7. The anti-proliferation activity is mediated by the arrest of the A549 and HepG2 cells in the S phase and MCF-7 in the G2/M phase of the cell cycle as suggested by flow cytometry. Results suggest that apoptosis and necrosis pathways ultimately lead to the death of the cancer cells.

In Vitro Anti-tumoral and Anti-bacterial Activity of an Octamolybdate Cluster-Based Hybrid Solid Incorporated with a Copper Picolinate Complex.

Inorganic drugs, especially polyoxometalate-based hybrids, are expected to be developed as promising future metallodrugs. Herein, an organic-inorganic hybrid solid based on pyridine-2-carboxylic acid or picolinic acid (pic), [(Cu(pic)2)2(Mo8O26)]·8H2O (1), was synthesized. A single-crystal structure of a solid possesses a discrete ß-type octamolybdate cluster that supramolecularly aggregates with a {Cu2(pic)4}4- complex and eight lattice water molecules. The study indicates that the solid is stable in aqueous medium and less toxic toward normal cell lines. The in vitro anti-bacterial and anti-tumor properties of the solid 1 were investigated. The results of the anti-tumor action against various human cancer cell lines, namely, lung (A549), breast (MCF-7), and liver (HepG2) cancer cells suggest that this ß-octamolybdate-based solid yielded the lowest IC50 value reported so far among octamolybdate anion-based hybrid solids, i.e., 24.24 µM for MCF-7, 21.56 µM for HepG2, and 25 µM for A549, indicating significant anti-cancer activity. The cell cycle analysis further reveals the observed anti-tumor effect to be governed by the arrest of breast cancer cells in the G2/M phase while that of lung and liver cancer cells in the S phase of the cell cycle. A fluorescence quenching study suggests the binding interaction between solid and ctDNA, which in turn induces apoptosis and necrosis pathways leading to cancer cell death. This is also the first study of {Mo8O26}4- cluster-based solids as an anti-bacterial agent against Escherichia coli, and it was found to be very effective with a minimal inhibitory concentration value of ∼135 µg/mL, which is the lowest so far reported for any octamolybdate-based solid.

Occult cobalamin and folate deficiency in Indians.

BACKGROUND: Our aim was to assess cobalamin and folate levels in normal Indian subjects before undertaking a prospective study of megaloblastic anaemia. METHODS: We took samples from 25 men and 25 women to establish the normal range. The exclusion criteria for subjects were age below 18 years and above 65 years, haemoglobin < 12 g/dl, and those who were pregnant, lactating or on any medication including vitamin supplements. A complete blood count and blood film examination for hypersegmented neutrophils were done. Serum cobalamin and folate assays were performed by a competitive immunoassay. The reference range supplied with the kits for serum cobalamin was 100-700 pg/ml and for serum folate it was 3-22 ng/ml. RESULTS: Since many 'normal' subjects in the sample showed values below the normal reference range, the numbers sampled were increased to 46 men and 50 women. Of all the subjects tested, 46.9% had subnormal values of one of the two vitamins. The normal ranges for serum cobalamin established in this study were--men 100-388 pg/ml and women 105.3-434 pg/ml. Of the 46 men tested, 17 (36.9%) had low cobalamin levels and of the 50 women tested, 23 (46%) had low cobalamin levels. Levels < 50 pg/ml were seen in 46.9% of these subjects. The normal ranges for serum folate in the study were--men 3.1 to > 22 ng/ml, women 3-12.26 ng/ml. In the study group, 8 men (17.3%) and 6 women (12%) had folate deficiency. Eight subjects (17%) had combined deficiency of the two vitamins. The mean corpuscular volume was not informative and was elevated in only 1 subject. Hypersegmentation of neutrophils was present in 75% of deficient subjects. CONCLUSION: We established normal levels for serum cobalamin and folate in our study group. Of the subjects studied, 46.9% had subnormal levels of serum cobalamin or serum folate, cobalamin deficiency being five times more common than folate. Hypersegmentation of neutrophils was a better indicator of occult megaloblastosis than the mean corpuscular volume.