Structural study of crystalline auromomycin: Preliminary X-ray diffraction study of auromomycin and macromomycin

Auromomycin and macromomycin from the organism Streptomyces macromomyceticus have been crystallized. The X-ray diffraction pattern of crystals of each molecule is consistent with space group P2₁2₁2 with cell parameters $\text{a = 46.45}\text{A}\text{?}\text{, b = 54.34}\text{A}\text{?}\text{and}\text{c = 42.03}\text{A}\text{?}$ for auromomycin, and $\text{a = 46.45}\text{A}\text{?}\text{, b = 54.52}\text{A}\text{?}\text{and}\text{c = 41.54}\text{A}\text{?}$ for macromomycin. Diffraction analysis of auromomycin is in progress.     

Physicochemical and biological properties of glycoproteins isolated from the plasma of different species of animals

1. Glycoproteins were isolated from the plasma of sheep, goat, cow, buffalo and monkey. They were homogeneous by electrophoresis; on ultracentrifugation, a faster-sedimenting fraction, to an extent of 5–8% only, was observed in each case. 2. Similar physical properties were exhibited by these glycoproteins and they each have a molecular weight of about 105000. 3. In chemical composition, differences have been observed and the glycoproteins can be classified into three groups: (a) sheep and goat glycoproteins; (b) cow and buffalo glycoproteins; (c) monkey glycoprotein. Glucose, galactosamine and N-terminal amino acid were absent from these proteins. 4. These glycoproteins were trypsin inhibitors and prolonged the clotting time of plasma.     

Les ostracodes neogenes du bassin de Savigne-sur-Lathan(Faluns de Touraine) I. Biostratigraphie et paleoecologie

The Neogen formations from the basin of Savigné-su-Lathan (Indre-et-Loire) have yielded 53 species of ostracods.The connections with the best known fauna from the Aquitaine basin and the Rhône basin are discussed.     

Neocarzinostatin: an antitumor protein a preliminary x-ray diffraction study.

The antitumor protein, neocarzinostatin, has been crystallized and examined by X-ray diffraction. Crystals of this globular protein are of space group P2₁2₁2₁ with cell parameters a = 27.4Å, b = 33.9Åand c = 102.0Å. There is one molecule of approximately 27Ådiameter per asymmetric unit. Crystals soaked in a K₂HgI₄ solution give diffraction patterns significantly different from native crystal diffraction patterns.     

Multiple targets of miR-302 and miR-372 promote reprogramming of human fibroblasts to induced pluripotent stem cells

The embryonic stem cell-specific cell cycle-regulating (ESCC) family of microRNAs (miRNAs) enhances reprogramming of mouse embryonic fibroblasts to induced pluripotent stem cells. Here we show that the human ESCC miRNA orthologs hsa-miR-302b and hsa-miR-372 promote human somatic cell reprogramming. Furthermore, these miRNAs repress multiple target genes, with downregulation of individual targets only partially recapitulating the total miRNA effects. These targets regulate various cellular processes, including cell cycle, epithelial-mesenchymal transition (EMT), epigenetic regulation and vesicular transport. ESCC miRNAs have a known role in regulating the unique embryonic stem cell cycle. We show that they also increase the kinetics of mesenchymal-epithelial transition during reprogramming and block TGFβ-induced EMT of human epithelial cells. These results demonstrate that the ESCC miRNAs promote dedifferentiation by acting on multiple downstream pathways. We propose that individual miRNAs generally act through numerous pathways that synergize to regulate and enforce cell fate decisions.     

Prevalence,antimicrobial resistance profile,and characterization of multi-drug resistant bacteria from various infected wounds in North Egypt

Multi-drug resistant (MDR) bacteria associated with wounds are extremely escalating. This study aims to survey different wounds in Alexandria hospitals, North Egypt, to explore the prevalence and characteristics of MDR bacteria for future utilization in antibacterial wound dressing designs. Among various bacterial isolates, we determined 22 MDR bacteria could resist different classes of antibiotics. The collected samples exhibited the prevalence of mono-bacterial infections (60%), while 40% included poly-bacterial species due to previous antibiotic administration. Moreover, Gram-negative bacteria showed dominance with a ratio of 63.6%, while Gram-positive bacteria reported 36.4%. Subsequently, the five most virulent bacteria were identified following the molecular approach by 16S rRNA and physiological properties using the VITEK 2 automated system. They were deposited in GenBank as Staphylococcus haemolyticus MST1 ({"type":"entrez-nucleotide","attrs":{"text":"KY550377","term_id":"1304487819","term_text":"KY550377"}}KY550377), Pseudomonas aeruginosa MST2 ({"type":"entrez-nucleotide","attrs":{"text":"KY550378","term_id":"1304487820","term_text":"KY550378"}}KY550378), Klebsiella pneumoniae MST3 ({"type":"entrez-nucleotide","attrs":{"text":"KY550379","term_id":"1304487821","term_text":"KY550379"}}KY550379), Escherichia coli MST4 ({"type":"entrez-nucleotide","attrs":{"text":"KY550380","term_id":"1304487822","term_text":"KY550380"}}KY550380), and Escherichia coli MST5 ({"type":"entrez-nucleotide","attrs":{"text":"KY550381","term_id":"1304487823","term_text":"KY550381"}}KY550381). In terms of isolation source, S. haemolyticus MST1 was isolated from a traumatic wound, while P. aeruginosa MST2 and E. coli MST4 were procured from hernia surgical wounds, and K. pneumoniae MST3 and E. coli MST5 were obtained from diabetic foot ulcers. Antibiotic sensitivity tests exposed that K. pneumoniae MST3, E. coli MST4, and E. coli MST5 are extended-spectrum β-lactamases (ESBLs) bacteria. Moreover, S. haemolyticus MST1 belongs to the methicillin-resistant coagulase-negative staphylococcus (MRCoNS), whereas P. aeruginosa MST2 exhibited resistance to common empirical bactericidal antibiotics. Overall, the study provides new insights into the prevalent MDR bacteria in Egypt for further use as specific models in formulating antibacterial wound dressings.     

Fluorescent adducts of wheat calmodulin implicate the amino-terminal region in the activation of skeletal muscle myosin light chain kinase

Considerable attention is being directed toward defining a binding site in the central region of calmodulin that forms a high affinity interaction with certain enzymes and amphiphilic peptides. However, other regions of calmodulin are also known to be involved in the activation of enzymes such as myosin light chain kinase, regions which may not be directly involved in the binding of small peptides, e.g. mastoparan X. We investigated the properties of wheat calmodulin fluorescent derivatives, which were modified chemically in the first calcium binding site at Cys-27, in the activation of rabbit fast skeletal muscle myosin light chain kinase. Unmodified wheat calmodulin stimulated myosin light chain kinase to a greater maximal velocity than wheat calmodulin that was modified at Cys-27 by any of four fluorescent compounds, IAANS (2-[4'-iodoacetamidoanilino]naphthalene-6-sulfonic acid), 5-[2'-[[iodoacetyl]amino]ethyl]aminonaphthalene]-1-sulfonic acid, 5-iodoacetamidofluorescein, and 7-diethylamino-3-[4'-maleimidylphenyl]-4-methylcoumarin; the midpoints for activation of myosin light chain kinase were not significantly different for unmodified wheat calmodulin and three of the four wheat calmodulin derivatives. Myosin light chain kinase, but not mastoparan X, enhanced the fluorescence emission intensity of wheat calmodulin-IAANS. Mastoparan X reversed, in a dose-dependent manner, the changes in fluorescence intensity of a preformed complex of myosin light chain kinase and wheat calmodulin-IANNS. Thus, we propose that the region vicinal to Cys-27 participates in the activation but not the high affinity association of myosin light chain kinase. Lastly, a comparison of mammalian and plant calmodulin showed that the Vmax for the stimulation of myosin light chain kinase was 1.6-fold greater for bovine than wheat calmodulin. The difference between the two calmodulins was more pronounced at lower Ca2+ because less Ca2+ was needed to saturate the kinase rate when stimulated by bovine calmodulin.     

Improvement of Intracellular Traffic System by Overexpression of KDEL Receptor 1 in Antibody‐Producing CHO Cells

The localization of soluble endoplasmic reticulum (ER) chaperones in the cell organelle is mediated by the C‐terminal KDEL (lysine, aspartic acid, glutamic acid and leucine) motif. This motif is recognized by the KDEL receptor, a seven‐transmembrane protein that cycles between the ER and cis‐Golgi to capture missorted KDEL chaperones from post‐ER compartments in a pH‐dependent manner. The KDEL receptor's target chaperones have a substantial role in protein folding and assembly. In this study, the gene expression level of KDEL receptor 1 shows a moderate upregulation during either ER stress or growth of Chinese hamster ovary (CHO) cells in batch culture, while the ER chaperones show higher upregulation. This might indicate the possibility of saturation of the ER retention machinery or at least hindered retention during late stage batch culture in recombinant CHO cells. KDELR1 is overexpressed in a monoclonal antibody‐producing CHO cell line to improve the intracellular chaperone retention rate in the ER. An increase in the specific productivity of IgG1 by 13.2% during the exponential phase, and 23.8% in the deceleration phase of batch culture is observed. This is the first study to focus on the ER retention system as a cell engineering target for enhancing recombinant protein production.