Synthesis, biological evaluation and docking studies of (4-aryl-3-methyl-4,5-dihydropyrazolo[3,4-c]pyrazol-1(3aH)-yl)(3-hydroxyquinoxalin-2-yl)methanones

862-866An efficient synthesis of (4-aryl-3-methyl-4,5-dihydropyrazolo[3,4-c]pyrazol-1(3aH)-yl)(3-hydroxyquinoxalin-2-yl) methanones is described via reactions of ethyl acetoacetate with 3-hydroxyquinoxaline-2-carbohydrazide. The structures of the compounds prepared have been determined by spectral analyses

Synthesis, biological evaluation and docking studies of (4-aryl-3-methyl-4,5-dihydropyrazolo[3,4-c]pyrazol-1(3aH)-yl)(3-hydroxyquinoxalin-2-yl)methanones

An efficient synthesis of (4-aryl-3-methyl-4,5-dihydropyrazolo[3,4-c]pyrazol-1(3aH)-yl)(3-hydroxyquinoxalin-2-yl) methanones is described via reactions of ethyl acetoacetate with 3-hydroxyquinoxaline-2-carbohydrazide. The structures of the compounds prepared have been determined by spectral analyses

Fungal Chitin Dampens Inflammation through IL-10 Induction Mediated by NOD2 and TLR9 Activation

Funding: JW and NARG thank the Wellcome Trust (080088, 086827, 075470), The Wellcome Trust Strategic Award in Medical Mycology and Fungal Immunology (097377) and the European Union ALLFUN (FP7/2007 2013, HEALTH-2010-260338) for funding. MGN was supported by a Vici grant of the Netherlands Organisation for Scientific Research. AJPB and DMM were funded by STRIFE, ERC-2009-AdG-249793 and AJPB additionally by FINSysB, PITN-GA-2008-214004 and the BBSRC [BB/F00513X/1]. MDL was supported by the MRC (MR/J008230/1). GDB and SV were funded by the Wellcome Trust (086558) and TB and MK were funded by the Deutsche Forschungsgemeinschaft (Bi 696/3-1; Bi 696/5-2; Bi 696/10-1). MS was supported by the Deutsche Forschungsgemeinschaft (Sch 897/1-3) and the National Institute of Dental and Craniofacial Research (R01 DE017514-01). TDK and RKSM were funded by the National Institute of Health (AR056296, AI101935) and the American Lebanese Syrian Associated Charities (ALSAC). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Peer reviewedPublisher PD

DFT Analysis, ADME, antibacterial activity and molecular docking studies of 2-(3-aryl-1,2,4-oxadiazol-5-yl)-n-phenylacetamide derivatives

Quantum computational study based on density functional theory (DFT/B3LYP) using basis set 6-311G (d,p) a number of global and local reactivity descriptors have been computed to predict the reactivity and the reactive sites on the 2-(3-aryl-1,2,4-oxadiazol-5-yl)-n-phenylacetamideoxadiazole derivatives. The molecular geometry and the electronic properties such as frontier molecular orbital (HOMO and LUMO), ionization potential (I) and electron affinity (A) are investigated to get a better insight of the molecular properties. Molecular electrostatic potential (MEP) for all compounds were determined to check their electrophilic or nucleophilic reactivity. The in silico pharmacokinetics showed that nearly all derivatives obeyed Lipinski rule of 5 with low toxicity and metabolic stability. The antibacterial activity was carried out against B. subtilis, S. aureus, P.aeruginosa and E. coli, displaying considerable inhibition. MurE ligases, (PDB: 7b6k) participating in the intracellular steps of bacterial peptidoglycan biosynthesis, are taken as targets for molecular docking studies using Flare GUI software. The docking outcome revealed that these 1,2,4-oxadiazole analogues have highest LF rank score in the range -12.9 to -6.0 which shows that they act as potent antibacterial agents

Isolation and Characterization of Two PHA Producing Strains Isolated from Soil Sample of Nalgonda District

Two whitish-colored gram-positive strains producing Polyhydroxyalkanoates (PHAs) were isolated from a soil sample from Nalgonda district in the Telangana state of India. With the help of Sudan black B staining, ten isolates with lipids, which showed bluish black color were selected from twenty-six bacterial strains which were selected randomly and purified from the serial diluted plate. Among ten isolates, 3D1 and 3D10 isolates were confirmed with Nile blue A and Nile Red staining for their PHA granules producing capacity. These two isolates grew optimally at a temperature of 37°C and a pH of 9. Furthermore, these strains were able to resist NaCl up to 10%, whereas, optimum NaCl required for the growth of 3D1 was 2%, but optimum NaCl required for the growth of 3D10 was shown to be 4%. PHAs produced by the two strains, 3D1 and 3D10, were extracted and quantified, which produced 68% PHA with a polymer concentration of 4.902 g/L and 61% PHA with a polymer concentration of 4.023 g/L, respectively. Biochemical, Morphological and Molecular characterization were performed on these two isolates. These two strains, 3D1 and 3D10, were closely related to Bacillus sonorensis with similarity of 99.51% and Bacillus safensis subsp. safensis with similarity of 99.66%, respectively. The 16S rDNA gene sequences of these two isolates were submitted to the NCBI Gene bank and the accession numbers were also sought

Aflatoxins B1 in different grades of chillies (Capsicum annum L.) in India as determined by indirect competitive-ELISA

Samples of the three grades of chilli pod (grades 1 to 3) were collected during surveys in 1998 and 1999 from the principal market yards and cold storage facilities of the major chilli-growing areas of Andhra Pradesh (AP), India. Chilli powders were collected from different supermarkets in Hyderabad, AP. They were analysed for aflatoxin B1 (AFB1) content by an indirect competitive ELISA. To avoid the influence of interfering substances present in chilli extracts, it was necessary to prepare the aflatoxin standards in methanol extracts of chillies free from aflatoxins. For nine representative samples there was good agreement between ELISA and HPLC estimations of AFB1 and the results suggested that the ELISA procedure adopted was dependable. Of the 182 chilli samples tested, 59% of the samples were contaminated with AFB1 and 18% contained the toxin at non-permissible levels. The highest AFB1 concentration of 969 µg/kg was found in one sample representing grade 3. Overall the maximum percentage of chilli pods showing AFB1 levels higher than 30 μg/kg (non-permissible levels) was in grade 3. Chilli pods stored in refrigerated rooms showed the lowest proportion of samples containing aflatoxin. Nearly 9% of the chilli powders sold in supermarkets contained non-permissible aflatoxin levels. This report highlights the importance of using grade 1 chilli pods to minimize aflatoxin contamination

Natural occurrence of ochratoxin A contamination in commercial black and white pepper products.

The concentration of ochratoxin A (OTA) in 120 commercial pepper (84 pre-packed and 36 bulk samples), which consist of local and imported white and black pepper in powder and seed form in Malaysia were determined. The objective of the study was to investigate and compare OTA concentration in black pepper and white pepper being commercialized in Malaysia. Determination method was based on HPLC with fluorescence detection coupled with immunoaffinity column clean-up step. Mobile phase consisted of acetonitrile-water-acetic acid (49.5:49.5:1.0, v/v/v), and flow rate was 1 ml/min. The LOD was 0.02 ng/g, and the average recovery values of OTA ranged from 79.5 to 92.0% in black pepper and 81.2-90.3% in white pepper. A total of 57 samples (47.5%) were contaminated with OTA ranging from 0.15 to 13.58 ng/g. The results showed that there was a significant difference between type of pepper and brands. OTA concentration in black pepper was significantly higher than white pepper (p < 0.05). The highest concentration of ochratoxin, 13.58 ng/g, was detected in a sample of black pepper seed followed by 12.64 ng/g in a sample of black pepper powder, both were bulk samples purchased from open market

Deletion of hematopoietic Dectin-2 or CARD9 does not protect against atherosclerotic plaque formation in hyperlipidemic mice

Inflammatory reactions activated by pattern recognition receptors (PRRs) on the membrane of innate immune cells play an important role in atherosclerosis. Whether the PRRs of the C-type lectin receptor (CLR) family including Dectin-2 may be involved in the pathogenesis of atherosclerosis remains largely unknown. Recently, the CLR-adaptor molecule caspase recruitment domain family member 9 (CARD9) has been suggested to play a role in cardiovascular pathologies as it provides the link between CLR activation and transcription of inflammatory cytokines as well as immune cell recruitment. We therefore evaluated whether hematopoietic deletion of Dectin-2 or CARD9 reduces inflammation and atherosclerosis development. Low-density lipoprotein receptor (Ldlr)- knockout mice were transplanted with bone marrow from wild-type, Dectin-2- or Card9-knockout mice and fed a Western-type diet containing 0.1% (w/w) cholesterol. After 10 weeks, lipid and inflammatory parameters were measured and atherosclerosis development was determined. Deletion of hematopoietic Dectin-2 or CARD9 did not influence plasma triglyceride and cholesterol levels. Deletion of hematopoietic Dectin-2 did not affect atherosclerotic lesion area, immune cell composition, ex vivo cytokine secretion by peritoneal cells or bone marrow derived macrophages. Unexpectedly, deletion of hematopoietic CARD9 increased atherosclerotic lesion formation and lesion severity. Deletion of hematopoietic CARD9 did also not influence circulating immune cell composition and peripheral cytokine secretion. Besides a tendency to a reduced macrophage content within these lesions, plasma MCP-1 levels decreased upon WTD feeding. Deletion of hematopoietic Dectin-2 did not influence atherosclerosis development in hyperlipidemic mice. The absence of CARD9 unexpectedly increased atherosclerotic lesion size and severity, suggesting that the presence of CARD9 may protect against initiation of atherosclerosis development

Identification and characterization of a large source of primary mesenchymal stem cells tightly adhered to bone surfaces of human vertebral body marrow cavities

Background: Therapeutic allogeneic mesenchymal stromal cells (MSCs) are currently in clinical trials to evaluate their effectiveness in treating many different disease indications. Eventual commercialization for broad distribution will require further improvements in manufacturing processes to economically manufacture MSCs at scales sufficient to satisfy projected demands. A key contributor to the present high cost of goods sold for MSC manufacturing is the need to create master cell banks from multiple donors, which leads to variability in large-scale manufacturing runs. Therefore, the availability of large single donor depots of primary MSCs would greatly benefit the cell therapy market by reducing costs associated with manufacturing. Methods: We have discovered that an abundant population of cells possessing all the hallmarks of MSCs is tightly associated with the vertebral body (VB) bone matrix and only liberated by proteolytic digestion. Here we demonstrate that these vertebral bone-adherent (vBA) MSCs possess all the International Society of Cell and Gene Therapy-defined characteristics (e.g., plastic adherence, surface marker expression and trilineage differentiation) of MSCs, and we have therefore termed them vBA-MSCs to distinguish this population from loosely associated MSCs recovered through aspiration or rinsing of the bone marrow compartment. Results: Pilot banking and expansion were performed with vBA-MSCs obtained from 3 deceased donors, and it was demonstrated that bank sizes averaging 2.9 × 108 ± 1.35 × 108 vBA-MSCs at passage 1 were obtainable from only 5 g of digested VB bone fragments. Each bank of cells demonstrated robust proliferation through a total of 9 passages, without significant reduction in population doubling times. The theoretical total cell yield from the entire amount of bone fragments (approximately 300 g) from each donor with limited expansion through 4 passages is 100 trillion (1 × 1014) vBA-MSCs, equating to over 105 doses at 10 × 106 cells/kg for an average 70-kg recipient. Discussion: Thus, we have established a novel and plentiful source of MSCs that will benefit the cell therapy market by overcoming manufacturing and regulatory inefficiencies due to donor-to-donor variability