Inositol‐requiring enzyme‐1 regulates phosphoinositide signaling lipids and macrophage growth

The ER‐bound kinase/endoribonuclease (RNase), inositol‐requiring enzyme‐1 (IRE1), regulates the phylogenetically most conserved arm of the unfolded protein response (UPR). However, the complex biology and pathology regulated by mammalian IRE1 cannot be fully explained by IRE1’s one known, specific RNA target, X box‐binding protein‐1 (XBP1) or the RNA substrates of IRE1‐dependent RNA degradation (RIDD) activity. Investigating other specific substrates of IRE1 kinase and RNase activities may illuminate how it performs these diverse functions in mammalian cells. We report that macrophage IRE1 plays an unprecedented role in regulating phosphatidylinositide‐derived signaling lipid metabolites and has profound impact on the downstream signaling mediated by the mammalian target of rapamycin (mTOR). This cross‐talk between UPR and mTOR pathways occurs through the unconventional maturation of microRNA (miR) 2137 by IRE1’s RNase activity. Furthermore, phosphatidylinositol (3,4,5) phosphate (PI(3,4,5)P₃) 5‐phosphatase‐2 (INPPL1) is a direct target of miR‐2137, which controls PI(3,4,5)P₃ levels in macrophages. The modulation of cellular PI(3,4,5)P₃/PIP₂ ratio and anabolic mTOR signaling by the IRE1‐induced miR‐2137 demonstrates how the ER can provide a critical input into cell growth decisions.     

Gene expression analysis of intestinal IL-8, IL-17 A and IL-10 in patients with celiac and inflammatory bowel diseases

Molecular Biology Reports - Celiac disease (CeD) and inflammatory bowel disease (IBD) are accompanied by impaired immune responses. To study the immune regulation of these diseases, we evaluated...     

Oligo-riboprobes. Tools for in situ hybridisation

In situ hybridisation detection of mRNAs using riboprobes has become a widely used technique. However, the identification of cells producing closely-related yet distinct mRNAs is difficult with the usual size probes. Moreover, it is not always easy to obtain the required cDNA essential for cRNA probe synthesis. To avoid these problems, we have used synthetic oligodeoxynucleotides to generate short, single stranded RNA probes ("oligo-riboprobes"). These probes can be labelled to very high (10(9) cpm/micrograms) specific activity and can be prepared for any published nucleotide sequence. We have used these probes to localise beta (preprotachykinin) PPT mRNA producing neurons in rat hypothalamus and bowel. The results were compared to that obtained with cRNA probes generated from beta preprotachykinin cDNA.     

Mitigating effect of nano-zerovalent iron,iron sulfate and EDTA against oxidative stress induced by chromium in <Emphasis Type="Italic">Helianthus annuus</Emphasis> L.

Due to its wide industrial application, chromium (Cr) is known to be a critical environmental pollutant. Contamination of water and agricultural soil by Cr inhibits crop productivity and their physiological and biochemical processes. The objective of the current work was to investigate the effects of appropriate reducing agents such as EDTA, iron sulfate (Fe²⁺), and zerovalent nano iron (Fe⁰ nanoparticles) on growth and physiology of sunflower plants under Cr(VI) stress. Results showed that the Cr uptake increased by increasing the amount of EDTA, leading to a significant reduction in morphological and physiological parameters except for MDA and H₂O₂ contents. Treatment with Fe⁰ nanoparticles and Fe²⁺ reduced Cr concentration in root and shoot, increased root and shoot dry weight, plastid pigments (chlorophyll and carotenoids) and proline contents; however, the level of MDA and H₂O₂ decreased significantly. All parameters were affected by Fe²⁺ during the first week of sampling; however, Fe⁰ nanoparticles affected all traits until the end of the third sampling stage. A statistically significant and positive correlation was found between root Cr concentration and MDA and H₂O₂ seedlings treated with EDTA, Fe²⁺, and Fe⁰ grown under Cr stress. From the result of this study, it can be concluded that sunflower has the potential for accumulation of Cr as a heavy metal, and treatment with Fe⁰ nanoparticles to prevent Cr uptake is more effective than other employed treatments.     

Generalized Baum-Welch Algorithm Based on the Similarity between Sequences

The profile hidden Markov model (PHMM) is widely used to assign the protein sequences to their respective families. A major limitation of a PHMM is the assumption that given states the observations (amino acids) are independent. To overcome this limitation, the dependency between amino acids in a multiple sequence alignment (MSA) which is the representative of a PHMM can be appended to the PHMM. Due to the fact that with a MSA, the sequences of amino acids are biologically related, the one-by-one dependency between two amino acids can be considered. In other words, based on the MSA, the dependency between an amino acid and its corresponding amino acid located above can be combined with the PHMM. For this purpose, the new emission probability matrix which considers the one-by-one dependencies between amino acids is constructed. The parameters of a PHMM are of two types; transition and emission probabilities which are usually estimated using an EM algorithm called the Baum-Welch algorithm. We have generalized the Baum-Welch algorithm using similarity emission matrix constructed by integrating the new emission probability matrix with the common emission probability matrix. Then, the performance of similarity emission is discussed by applying it to the top twenty protein families in the Pfam database. We show that using the similarity emission in the Baum-Welch algorithm significantly outperforms the common Baum-Welch algorithm in the task of assigning protein sequences to protein families.     

Disruption of tubulin polymerization and cell proliferation by 1-naphthylarsonic acid

Arsenical compounds exhibit a differential toxicity to cancer cells. Microtubules are a primary target of a number of anticancer drugs, such as arsenical compounds. The interaction of 1-NAA (1-naphthylarsonic acid) has been investigated on microtubule polymerization under in vitro and cellular conditions. Microtubules were extracted from sheep brain. Transmission electron microscopy was used to show microtubule structure in the presence of 1-NAA. Computational docking method was applied for the discovery of ligand-binding sites on the microtubular proteins. Proliferation of HeLa cells and HF2 (human foreskin fibroblasts) was measured by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide] assay method following their incubation with 1-NAA. Fluorescence microscopic labelling was done with the help of α-tubulin monoclonal antibody and Tunel kit was used to investigate the apoptotic effects of 1-NAA on the HeLa cells. 1-NAA inhibits the tubulin polymerization by the formation of abnormal polymers having high affinity to the inner cell wall.     

The toxicity study of a systemic fungicide: artea 330 EC on the physiology and the respiratory metabolism of the tadpole (Rana saharica)

The intensification of the cereal cultures accompanied by the apparition of damaging illnesses for these cultures. These illnesses are Largely imputed to mushrooms micro and macroscopic chatty of important damages at wheat, the barley and of none targeted other animal species. The products used against these illnesses are called: Fungicides. In our work, we are interested in the survey of the effects of a systemic fungicide: the ARTEA 330 EC introduces newly in Algeria, on some physiological and metabolic parameters of a biologic model: the tadpole The exhibition of the populations of tadpoles to the different concentrations of the ARTEA 330 EC provokes a fall very important of the middleweights of the tadpoles treated. This one is reduced of meadows of 80% to the strongest concentration of ARTEA 330 EC. It is some in the same way for the reduction of the middle size of the tadpoles where one records a reduction of 25%. Concerning the percentage of mortality gotten, we observe that the one is here from 100% to the strongest concentration of ARTEA 330 EC. The breathing of the tadpoles treated by fungicide is disrupted strongly and to the strongest concentration, this one is inhibited completely. The gotten results show that the exhibition of the populations of tadpoles to the different concentrations of fungicide disrupts the physiological parameters strongly and inhibit the respiratory metabolism. A phenomenon of detoxication seems to intervene; it is put in evidence by a stimulation of the synthesis of the proteins.     

Induced in vitro differentiation of neural-like cells from human exfoliated deciduous teeth-derived stem cells

Stem cells from human exfoliated deciduous teeth (SHED) are highly proliferative, clonogenic and multipotent stem cells with a neural crest cell origin. Additionally, they can be collected with minimal invasiveness in comparison with other sources of mesenchymal stem cells (MSCs). Therefore, SHED could be a desirable option for potential therapeutic applications. In this study, SHEDs were established from enzyme-disaggregated deciduous dental pulp obtained from 6 to 9 year-old children. The cells had typical fibroblastoid morphology and expressed antigens characteristic of MSCs, STRO1, CD146, CD45, CD90, CD106 and CD166, but not the hematopoietic and endothelial markers, CD34 and CD31, as assessed by FACS analysis. Differentiation assessment revealed a strong osteogenic and adipogenic potential of SHEDs. In order to further evaluate the in vitro differentiation potential of SHED into neural cells, a simple short time growth factor-mediated induction was used. Immunofluorescence staining and flow cytometric analysis revealed that SHED rapidly expressed nestin and b-III tubulin, and later expressed intermediate neural markers. In addition, the intensity and percentages of nestin and b-III tubulin and mature neural markers (PSA-NCAM, NeuN, Tau, TH, or GFAP) increased significantly following treatment. Moreover, RT-PCR and Western blot analyses showed that the neural markers were strongly up-regulated after induction. In conclusion, these results provide evidence that SHED can differentiate into neural cells by the expression of a comprehensive set of genes and proteins that define neural-like cells in vitro. SHED cells might be considered as new candidates for the autologous transplantation of a wide variety of neurological diseases and neurotraumatic injuries.