The replacement of five consecutive amino acids in the cyt1a protein of bacillus thuringiensis enhances its cytotoxic activity against lung epithelial cancer cells

Cyt1A protein is a cytolytic protein encoded by the cyt gene of Bacillus thuringiensis subsp. israelensis (Bti) as part of the parasporal crystal proteins produced during the sporulation. Cyt1A protein is unique compared to the other endotoxins present in these parasporal crystals. Unlike ?-endotoxins, Cyt1A protein does not require receptors to bind to the target cell and activate the toxicity. It has the ability to affect a broad range of cell types and organisms, due to this characteristic. Cyt1A has been recognized to not only target the insect cells directly, but also recruit other endotoxins by acting as receptors. Due to these mode of actions, Cyt1A has been studied for its cytolytic activity against human cancer cell lines, although not extensively. In this study, we report a novel Cyt1A protein produced by a Bti strain QBT229 isolated from Qatar. When tested for its cytotoxicity against lung cancer cells, this local strain showed considerably higher activity compared to that of the reference Bti and other strains tested. The possible reasons for such enhanced activity were explored at the gene and protein levels. It was evidenced that five consecutive amino acid replacements in the ?8 sheet of the Cyt1A protein enhanced the cytotoxicity against the lung epithelial cancer cells. Such novel Cyt1A protein with high cytotoxicity against lung cancer cells has been characterized and reported through this study. -2018 by the authors. Licensee MDPI, Basel, Switzerland.Scopu

Greensporone A, a fungal secondary metabolite suppressed constitutively activated AKT via ROS generation and induced apoptosis in leukemic cell lines

Greensporone A is a fungal secondary metabolite that has exhibited potential in vitro for anti-proliferative activity in vitro. We studied the anticancer activity of greensporone A in a panel of leukemic cell lines. Greensporone A-mediated inhibition of proliferation is found to be associated with the induction of apoptotic cell death. Greensporone A treatment of leukemic cells causes inactivation of constitutively activated AKT and its downstream targets, including members GSK3 and FOXO1, and causes downregulation of antiapoptotic genes such as Inhibitor of Apoptosis (IAPs) and Bcl-2. Furthermore, Bax, a proapoptotic member of the Bcl-2 family, was found to be upregulated in leukemic cell lines treated with greensporone A. Interestingly, gene silencing of AKT using AKT specific siRNA suppressed the expression of Bcl-2 with enhanced expression of Bax. Greensporone A-mediated increase in Bax/Bcl-2 ratio causes permeabilization of the mitochondrial membrane leading to the accumulation of cytochrome c in the cytoplasm. Greensporone A-induced cytochrome c accumulation causes the activation of caspase cascade and cleavage of its effector, poly(ADP-ribose) polymerase (PARP), leading to apoptosis. Greensporone A-mediated apoptosis in leukemic cells occurs through the generation of reactive oxygen species (ROS) due to depletion of glutathione (GSH) levels. Finally, greensporone A potentiated the anticancer activity of imatinib in leukemic cells. In summary, our study showed that greensporone A suppressed the growth of leukemic cells via induction of apoptotic cell death. The apoptotic cell death occurs by inhibition of AKT signaling and activation of the intrinsic apoptotic/caspase pathways. These results raise the possibility that greensporone A could be developed as a therapeutic agent for the treatment of leukemia and other hematological malignancies.Qatar University , University of North Carolina, National Palliative Care Research Center, Jordan University of Science and Technolog

Fractal analysis of microCT images of the oviduct during the estrous cycle

It is well known that the oviduct plays a key role in several events deeply related with reproduction, such as sperm storage and capacitation, gametes interactions, fertilization and early embryo development, among others. To better understand some of the interactions and process occurring withing this organ, the study of its morphological modifications is of primordial importance. To that, we adopted a microtomografy (MicroTC) modelling system and the fractal analysis that allow to explore the 3D oviductal functional anatomy, by using eight swine oviducts at different stages of the estrous cycle. MicroCT datasets were acquired by using the high-resolution 3D-imaging system Skyscan 1172G (Bruker, Kontich – Belgium). CT images were analyzed using plugin on ImageJ software (NIH, Bethesda, MD), a box-counting method was applied to calculate the Fractal dimension of the oviduct. Focusing our attention on the utero-tubal junction (involved in sperm selection) and the isthmo-ampullar junction (the fertilization site). We found that by using PCA analysis it was possible to clearly differentiate the oviduct at different cycle stage on the basis of their values for: Db for grid, lacunarity for grid, R2 for Db, Media Db, lacunarity, σ for D for Db, Max for D, Min for D. Lacunarity, Media and Max for Db have a greater influence on the analysis. The results showed that 2 principal components were associated whit the morphological changes. This information, is obtained by a fast nondestructive method, and could be very useful because this innovative approach enables the achievement a 3D model and suggest that using fractal analysis techniques can aid to better understand the modifications of oviduct anatomy that depends on the neuroendocrine axis. This innovative approach could be a start point to design 3D cell culture systems, that could be applied in human and animal assisted reproductive techniques, improving the IVF outcomes

An integrated multi-omic approach demonstrates distinct molecular signatures between human obesity with and without metabolic complications: a case–control study

Objectives: To examine the hypothesis that obesity complicated by the metabolic syndrome, compared to uncomplicated obesity, has distinct molecular signatures and metabolic pathways. Methods: We analyzed a cohort of 39 participants with obesity that included 21 with metabolic syndrome, age-matched to 18 without metabolic complications. We measured in whole blood samples 754 human microRNAs (miRNAs), 704 metabolites using unbiased mass spectrometry metabolomics, and 25,682 transcripts, which include both protein coding genes (PCGs) as well as non-coding transcripts. We then identified differentially expressed miRNAs, PCGs, and metabolites and integrated them using databases such as mirDIP (mapping between miRNA-PCG network), Human Metabolome Database (mapping between metabolite-PCG network) and tools like MetaboAnalyst (mapping between metabolite-metabolic pathway network) to determine dysregulated metabolic pathways in obesity with metabolic complications. Results: We identified 8 significantly enriched metabolic pathways comprising 8 metabolites, 25 protein coding genes and 9 microRNAs which are each differentially expressed between the subjects with obesity and those with obesity and metabolic syndrome. By performing unsupervised hierarchical clustering on the enrichment matrix of the 8 metabolic pathways, we could approximately segregate the uncomplicated obesity strata from that of obesity with metabolic syndrome. Conclusions: The data suggest that at least 8 metabolic pathways, along with their various dysregulated elements, identified via our integrative bioinformatics pipeline, can potentially differentiate those with obesity from those with obesity and metabolic complications

SnoRNAs and miRNAs networks underlying COVID-19 disease severity

There is a lack of predictive markers for early and rapid identification of disease progression in COVID-19 patients. Our study aims at identifying microRNAs (miRNAs)/small nucleolar RNAs (snoRNAs) as potential biomarkers of COVID-19 severity. Using differential expression analysis of microarray data (n = 29), we identified hsa-miR-1246, ACA40, hsa-miR-4532, hsa-miR-145-5p, and ACA18 as the top five differentially expressed transcripts in severe versus asymptomatic, and ACA40, hsa-miR-3609, ENSG00000212378 (SNORD78), hsa-miR-1231, hsa-miR-885-3p as the most significant five in severe versus mild cases. Moreover, we found that white blood cell (WBC) count, absolute neutrophil count (ANC), neutrophil (%), lymphocyte (%), red blood cell (RBC) count, hemoglobin, hematocrit, D-Dimer, and albumin are significantly correlated with the identified differentially expressed miRNAs and snoRNAs. We report a unique miRNA and snoRNA profile that is associated with a higher risk of severity in a cohort of SARS-CoV-2 infected patients. Altogether, we present a differential expression analysis of COVID-19-associated microRNA (miRNA)/small nucleolar RNA (snoRNA) signature, highlighting their importance in SARS-CoV-2 infection

Characteristic MicroRNAs Linked to Dysregulated Metabolic Pathways in Qatari Adult Subjects With Obesity and Metabolic Syndrome

BackgroundObesity-associated dysglycemia is associated with metabolic disorders. MicroRNAs (miRNAs) are known regulators of metabolic homeostasis. We aimed to assess the relationship of circulating miRNAs with clinical features in obese Qatari individuals.MethodsWe analyzed a dataset of 39 age-matched patients that includes 18 subjects with obesity only (OBO) and 21 subjects with obesity and metabolic syndrome (OBM). We measured 754 well-characterized human microRNAs (miRNAs) and identified differentially expressed miRNAs along with their significant associations with clinical markers in these patients.ResultsA total of 64 miRNAs were differentially expressed between metabolically healthy obese (OBO) versus metabolically unhealthy obese (OBM) patients. Thirteen out of 64 miRNAs significantly correlated with at least one clinical trait of the metabolic syndrome. Six out of the thirteen demonstrated significant association with HbA1c levels; miR-331-3p, miR-452-3p, and miR-485-5p were over-expressed, whereas miR-153-3p, miR-182-5p, and miR-433-3p were under-expressed in the OBM patients with elevated HbA1c levels. We also identified, miR-106b-3p, miR-652-3p, and miR-93-5p that showed a significant association with creatinine; miR-130b-5p, miR-363-3p, and miR-636 were significantly associated with cholesterol, whereas miR-130a-3p was significantly associated with LDL. Additionally, miR-652-3p’s differential expression correlated significantly with HDL and creatinine.ConclusionsMicroRNAs associated with metabolic syndrome in obese subjects may have a pathophysiologic role and can serve as markers for obese individuals predisposed to various metabolic diseases like diabetes

Abyssal circulation in the Indian Ocean from a 1 / 12 ∘ resolution global hindcast

Abyssal pathways and volume transports in the Indian Ocean obtained from a 1 / 12 ∘ global data assimilative hindcast for the years 2004–2006 are presented. The known features of bottom water circulation such as boundary currents and inter-basin exchanges are well represented in the hindcast solution. Further, the hindcast solution reveals a basin-wide, anti-cyclonic deep circulation pattern with the main inflow located in the east, in the Perth Basin, and outflow in the west along the African continent and Madagascar. Inter-basin westward transport at 10 ∘ S across the Ninetyeast Ridge and along the Mascarene Plateau connect the inflow and outflow locations. The model develops a net bottom water inflow of 10 ± 8 Sv ( mean ± standard deviation ) across 32 ∘ S , below approximately 3500 m of which 4 ± 7 Sv are returned south across the same latitude in the deep layers between 2000 and 3500 m. Significant variability on intra-seasonal, semi-annual and annual time scales is present in the abyssal flow fields. The vertical mixing algorithm, based on resolved vertical shear, suggests enhanced diapycnal upwelling in the vicinity of the Central Indian Ridge

Wind driven general circulation of the Mediterranean Sea simulated with a Spectral Element Ocean Model

This work is an attempt to simulate the Mediterranean Sea general circulation with a Spectral Finite Element Model. This numerical technique associates the geometrical flexibility of the finite elements for the proper coastline definition with the precision offered by spectral methods. The model is reduced gravity and we study the wind-driven ocean response in order to explain the large scale sub-basin gyres and their variability. The study period goes from January 1987 to December 1993 and two forcing data sets are used. The effect of wind variability in space and time is analyzed and the relationship between wind stress curl and ocean response is stressed. Some of the main permanent structures of the general circulation (Gulf of Lions cyclonic gyre, Rhodes gyre, Gulf of Syrte anticylone) are shown to be induced by permanent wind stress curl structures. The magnitude and spatial variability of the wind is important in determining the appearance or disappearance of some gyres (Tyrrhenian anticyclonic gyre, Balearic anticyclonic gyre, Ionian cyclonic gyre). An EOF analysis of the seasonal variability indicates that the weakening and strengthening of the Levantine basin boundary currents is a major component of the seasonal cycle in the basin. The important discovery is that seasonal and interannual variability peak at the same spatial scales in the ocean response and that the interannual variability includes the change in amplitude and phase of the seasonal cycle in the sub-basin scale gyres and boundary currents. The Coriolis term in the vorticity balance seems to be responsible for the weakening of anticyclonic structures and their total disappearance when they are close to a boundary. The process of adjustment to winds produces a train of coastally trapped gravity waves which travel around the eastern and western basins, respectively in approximately 6 months. This corresponds to a phase velocity for the wave of about 1 m/s, comparable to an average velocity of an internal Kelvin wave in the area