Cloning and Over Expression of an Aspergillus niger XP Phytase Gene (phyA) in Pichia pastoris

A. niger XP isolated from Vietnam produces very low amount of acidic phytase with optimal pH at 2.5 and 5.5. The phytase production of this strain was successfully improved through gene cloning and expression. A 1.4 - kb DNA fragment containing the coding region of the phyA gene was amplified by PCR and inserted into the expression vector pPICZαA with a signal peptide α- factor, under the control of AOX1 promoter. The recombined plasmid was transformed into the host strain P. pastoris KM71H and X33 by electroporation. Both host strains could efficiently express and secret phytase. The multicopy strains were screened for over expression of phytase. All the selected multicopy strains of P. pastoris X33 were examined for phytase activity, the maximum phytase yield of 1329 IU/ml was obtained after 4 days of incubation in medium BMM. The recombinant protein with MW of 97.4 KW showed to be the only one protein secreted in the culture broth. Multicopy transformant P. pastoris X33 supposed to be potential candidate for producing the commercial preparation of phytase.

Left Ventricular Model to Study the Combined Viscoelastic, Heart Rate, and Size Effects

It is known that the heart interacts with and adapts to its venous and arterial loading conditions. Various experimental studies and modeling approaches have been developed to investigate the underlying mechanisms. This paper presents a model of the left ventricle derived based on nonlinear stress-length myocardial characteristics integrated over truncated ellipsoidal geometry, and second-order dynamic mechanism for the excitation-contraction coupling system. The results of the model presented here describe the effects of the viscoelastic damping element of the electromechanical coupling system on the hemodynamic response. Different heart rates are considered to study the pacing effects on the performance of the left-ventricle against constant preload and afterload conditions under various damping conditions. The results indicate that the pacing process of the left ventricle has to take into account, among other things, the viscoelastic damping conditions of the myofilament excitation-contraction process. The effects of left ventricular dimensions on the hemdynamic response have been examined. These effects are found to be different at different viscoelastic and pacing conditions.

Salicylhydroxamic Acid Inhibits the Growth of Candida albicans

Candida spp. are common and aggressive pathogens. Because of the growing resistance of Candida spp. to current antifungals, novel targets, found in Candida spp. but not in humans or other flora, have to be identified. The alternative oxidase (AOX) is one such possibility. This enzyme is insensitive to cyanide, but is sensitive to compounds such as salicylhydroxamic acid (SHAM), disulfiram and n-alkyl gallates. The growth Candida albicans was inhibited by SHAM (Ki = 9-15 mM) and cyanide (Ki = 2-4 mM), albeit to differing extents. The rate of O2 uptake was inhibited by less than 10% by 25 mM SHAM and by about 90% by 250 μM KCN. Although SHAM substantially inhibited the growth of C. albicans, it is unlikely that the inhibition of AOX was the cause. Salicylhydroxamic acid is used therapeutically in the treatment of urinary tract infections and urolithiasis, but it also has some potential in the treatment of C. albicans infection.

Acoustic Finite Element Analysis of a Slit Model with Consideration of Air Viscosity

In very narrow pathways, the speed of sound propagation and the phase of sound waves change due to the air viscosity. We have developed a new finite element method (FEM) that includes the effects of air viscosity for modeling a narrow sound pathway. This method is developed as an extension of the existing FEM for porous sound-absorbing materials. The numerical calculation results for several three-dimensional slit models using the proposed FEM are validated against existing calculation methods.

Cloning of a β-Glucosidase Gene (BGL1) from Traditional Starter Yeast Saccharomycopsis fibuligera BMQ 908 and Expression in Pichia pastoris

β-Glucosidase is an important enzyme for production of ethanol from lignocellulose. With hydrolytic activity on cellooligosaccharides, especially cellobiose, β-glucosidase removes product inhibitory effect on cellulases and forms fermentable sugars. In this study, β-glucosidase encoding gene (BGL1) from traditional starter yeast Saccharomycosis fibuligera BMQ908 was cloned and expressed in Pichia pastoris. BGL1 of S. fibuligera BMQ 908 shared 98% nucleotide homology with the closest GenBank sequence (M22475) but identity in amino-acid sequences of catalytic domains. Recombinant plasmid pPICZαA/BGL1 containing the sequence encoding BGL1 mature protein and α-factor secretion signal was constructed and transformed into methylotrophic yeast P. pastoris by electroporation. The recombinant strain produced single extracellular protein with molecular weight of 120 kDa and cellobiase activity of 60 IU/ml. The optimum pH of the recombinant β-glucosidase was 5.0 and the optimum temperature was 50°C.

The Role of Immunogenic Adhesin Vibrio alginolyticus 49 k Da to Molecule Expression of Major Histocompatibility Complex on Receptors of Humpback Grouper Cromileptes altivelis

The purpose of research was to know the role of immunogenic protein of 49 kDa from V.alginolyticus which capable to initiate molecule expression of MHC Class II in receptor of Cromileptes altivelis. The method used was in vivo experimental research through testing of immunogenic protein 49 kDa from V.alginolyticus at Cromileptes altivelis (size of 250 - 300 grams) using 3 times booster by injecting an immunogenic protein in a intramuscular manner. Response of expressed MHC molecule was shown using immunocytochemistry method and SEM. Results indicated that adhesin V.alginolyticus 49 kDa which have immunogenic character could trigger expression of MHC class II on receptor of grouper and has been proven by staining using immunocytochemistry and SEM with labeling using antibody anti MHC (anti mouse). This visible expression based on binding between epitopes antigen and antibody anti MHC in the receptor. Using immunocytochemistry, intracellular response of MHC to in vivo induction of immunogenic adhesin from V.alginolyticus was shown.

Ratio-Dependent Food Chain Models with Three Trophic Levels

In this paper we study a food chain model with three trophic levels and Michaelis-Menten type ratio-dependent functional response. Distinctive feature of this model is the sensitive dependence of the dynamical behavior on the initial populations and parameters of the real world. The stability of the equilibrium points are also investigated.

The Dynamics of Oil Bodies in A. thaliana Seeds: A Mathematical Model of Biogenesis and Coalescence

The subcellular organelles called oil bodies (OBs) are lipid-filled quasi-spherical droplets produced from the endoplasmic reticulum (ER) and then released into the cytoplasm during seed development. It is believed that an OB grows by coalescence with other OBs and that its stability depends on the composition of oleosins, major proteins inserted in the hemi membrane that covers OBs. In this study, we measured the OB-volume distribution from different genotypes of A. thaliana after 7, 8, 9, 10 and 11 days of seed development. In order to test the hypothesis of OBs dynamics, we developed a simple mathematical model using non-linear differential equations inspired from the theory of coagulation. The model describes the evolution of OB-volume distribution during the first steps of seed development by taking into consideration the production of OBs, the increase of triacylglycerol volume to be stored, and the growth by coalescence of OBs. Fitted parameters values show an increase in the OB production and coalescence rates in A. thaliana oleosin mutants compared to wild type.

Leatherback Turtle (Dermochelys coriacea) after Incubation Eggshell in Andaman Sea, Thailand Study: Microanalysis on Ultrastructure and Elemental Composition

There are few studies on eggshell of leatherback turtle which is endangered species in Thailand. This study was focusing on the ultrastructure and elemental composition of leatherback turtle eggshells collected from Andaman Sea Shore, Thailand during the nesting season using scanning electron microscope (SEM). Three eggshell layers of leatherback turtle; the outer cuticle layer or calcareous layer, the middle layer or middle multistrata layer and the inner fibrous layer were recognized. The outer calcareous layer was thick and porosity which consisted of loose nodular units of various crystal shapes and sizes. The loose attachment between these units resulted in numerous spaces and openings. The middle layer was compact thick with several multistrata and contained numerous openings connecting to both outer cuticle layer and inner fibrous layer. The inner fibrous layer was compact and thin, and composed of numerous reticular fibers. Energy dispersive X-ray microanalysis detector revealed energy spectrum of X-rays character emitted from all elements on each layer. The percentages of all elements were found in the following order: carbon (C) > oxygen (O) > calcium (Ca) > sulfur (S) > potassium (K) > aluminum (Al) > iodine (I) > silicon (Si) > chlorine (Cl) > sodium (Na) > fluorine (F) > phosphorus (P) > magnesium (Mg). Each layer consisted of high percentage of CaCO3 (approximately 98%) implying that it was essential for turtle embryonic development. A significant difference was found in the percentages of Ca and Mo in the 3layers. Moreover, transition metal, metal and toxic non-metal contaminations were found in leatherback turtle eggshell samples. These were palladium (Pd), molybdenum (Mo), copper (Cu), aluminum (Al), lead (Pb), and bromine (Br). The contamination elements were seen in the outer layers except for Mo. All elements were readily observed and mapped using Smiling program. X-ray images which mapped the location of all elements were showed. Calcium containing in the eggshell appeared in high contents and was widely distributing in clusters of the outer cuticle layer to form CaCO3 structure. Moreover, the accumulation of Na and Cl was observed to form NaCl which was widely distributing in 3 eggshell layers. The results from this study would be valuable on assessing the emergent success in this endangered species.

Identification and Analysis of Binding Site Residues in Protein-Protein Complexes

We have developed an energy based approach for identifying the binding sites and important residues for binding in protein-protein complexes. We found that the residues and residuepairs with charged and aromatic side chains are important for binding. These residues influence to form cation-¤Ç, electrostatic and aromatic interactions. Our observation has been verified with the experimental binding specificity of protein-protein complexes and found good agreement with experiments. The analysis on surrounding hydrophobicity reveals that the binding residues are less hydrophobic than non-binding sites, which suggests that the hydrophobic core are important for folding and stability whereas the surface seeking residues play a critical role in binding. Further, the propensity of residues in the binding sites of receptors and ligands, number of medium and long-range contacts, and influence of neighboring residues will be discussed.

Mammogram Image Size Reduction Using 16-8 bit Conversion Technique

Two algorithms are proposed to reduce the storage requirements for mammogram images. The input image goes through a shrinking process that converts the 16-bit images to 8-bits by using pixel-depth conversion algorithm followed by enhancement process. The performance of the algorithms is evaluated objectively and subjectively. A 50% reduction in size is obtained with no loss of significant data at the breast region.

The Effect of Guanidine Hydrochloride on Phase Diagram of PEG- Phosphate Aqueous Two-Phase System

This report focus on phase behavior of polyethylene glycol (PEG)4000/ phosphate/ guanidine hydrochloride/ water system at different guanidine hydrochloride concentrations and pH. The binodal of the systems was displaced toward higher concentrations of the components with increasing guanidine hydrochloride concentrations. The partition coefficient of guanidine hydrochloride was near unity and increased with decreasing pH and increasing PEG/salt (%w/w) ratio.

How the Kinematic Swimming of European Eel Anguilla Anguilla Changes from Axial to Non-axial Velocity Flow

The aim of this study is to investigate the kinematics of undulatory elongated fish swimming against a velocity flow. We perform the experiments on European eel Anguilla Anguilla swimming in a hydrodynamic re-circulating tank with the velocity flow fixed at 0.2 m/s. We find that the undulating shape of overall eel body changes when it swims slantwise from the flow direction, by comparison to axial undulation shape. We examine this kinematics and we propose a general equation describing the lateral position of undulation body taking into account the direction of the eel-s swimming.

Burst on Hurst Algorithm for Detecting Activity Patterns in Networks of Cortical Neurons

Electrophysiological signals were recorded from primary cultures of dissociated rat cortical neurons coupled to Micro-Electrode Arrays (MEAs). The neuronal discharge patterns may change under varying physiological and pathological conditions. For this reason, we developed a new burst detection method able to identify bursts with peculiar features in different experimental conditions (i.e. spontaneous activity and under the effect of specific drugs). The main feature of our algorithm (i.e. Burst On Hurst), based on the auto-similarity or fractal property of the recorded signal, is the independence from the chosen spike detection method since it works directly on the raw data.

Antioxydant and Antibacterial Activity of Alkaloids and Terpenes Extracts from Euphorbia granulata

In order to enhance the knowledge of certain phytochemical Algerian plants that are widely used in traditional medicine and to exploit their therapeutic potential in modern medicine, we have done a specific extraction of terpenes and alkaloids from the leaves of Euphorbia granulata to evaluate the antioxidant and antibacterial activity of this extracts. After the extraction it was found that the terpene extract gave the highest yield 59.72% compared with alkaloids extracts. The disc diffusion method was used to determine the antibacterial activity against different bacterial strains: Escherichia coli (ATCC25922), Pseudomonas aeruginosa (ATCC27853) and Staphylococcus aureus (ATCC25923). All extracts have shown inhibition of growth bacteria. The different zones of inhibition have varied from (7 -10 mm) according to the concentrations of extract used. Testing the antiradical activity on DPPH-TLC plates indicated the presence of substances that have potent anti-free radical. As against, the BC-TLC revealed that only terpenes extract which was reacted positively. These results can validate the importance of Euphorbia granulata in traditional medicine.

The Identification of Anuran Glial Cells

Attempts were made to identify anuran glial cells. They were found as nervous tissue resident. Having stage dependent morphotype changes, whereby, appeared as an ovoid to oval in resting state and amoeboid mrophotypes in activated state, stained fairly with methylene blue and take up Pelikane blue 10% aqueous solution, as well as having the ability to phagocytize heat killed Staphylococcus aureus. They were delineated from the migrating peripheral monocytes by morphotypic and morphometeric differences. Such criteria were consistence with glial cells. Thus, the anuran glial cells are being identified in the frog Rana ridibunda Pallas 1771 and this animal can be of use as a simple model for the immunobiology of glial cells.

Histological Study of Postmortem Juvenile Green Sea Turtle (Chelonia mydas) from Royal Thai Navy Sea Turtle Nursery, Phang-nga, Thailand

The problem on the conservation programme of the Royal Thai Navy Sea Turtle Nursery, Phang-nga Province, Thailand is high mortality rate of juvenile green sea turtle (Cheloniamydas) on nursing period. So, during May to October 2012, postmortem examinations of juvenile green sea turtle were performed to determine the causes of dead. Fresh tissues of postmortem of 15 juvenile green sea turtles (1-3 months old) were investigated using paraffin section technique. The results showed normal ultrastructure of all tissue organs. These instances reviewed the health and stability of the environments in which juvenile green sea turtles live and concern for the survival rate. The present article also provides guidance for a review of the biology, guidelines for appropriate postmortem tissue, normal histology and sampling collection and procedures. The data also provides information for conservation of this endangered species in term of acknowledging and encouraging people to protect the animals and their habitats in nature.

Statistics of Exon Lengths in Animals, Plants, Fungi, and Protists

Eukaryotic protein-coding genes are interrupted by spliceosomal introns, which are removed from the RNA transcripts before translation into a protein. The exon-intron structures of different eukaryotic species are quite different from each other, and the evolution of such structures raises many questions. We try to address some of these questions using statistical analysis of whole genomes. We go through all the protein-coding genes in a genome and study correlations between the net length of all the exons in a gene, the number of the exons, and the average length of an exon. We also take average values of these features for each chromosome and study correlations between those averages on the chromosomal level. Our data show universal features of exon-intron structures common to animals, plants, and protists (specifically, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster, Cryptococcus neoformans, Homo sapiens, Mus musculus, Oryza sativa, and Plasmodium falciparum). We have verified linear correlation between the number of exons in a gene and the length of a protein coded by the gene, while the protein length increases in proportion to the number of exons. On the other hand, the average length of an exon always decreases with the number of exons. Finally, chromosome clustering based on average chromosome properties and parameters of linear regression between the number of exons in a gene and the net length of those exons demonstrates that these average chromosome properties are genome-specific features.

Utilization of 3-N-trimethylamino-1-propanol by Rhodococcus sp. strain A4 isolated from Natural Soil

The aim of this study was to screen for microorganism that able to utilize 3-N-trimethylamino-1-propanol (homocholine) as a sole source of carbon and nitrogen. The aerobic degradation of homocholine has been found by a gram-positive Rhodococcus sp. bacterium isolated from soil. The isolate was identified as Rhodococcus sp. strain A4 based on the phenotypic features, physiologic and biochemical characteristics, and phylogenetic analysis. The cells of the isolated strain grown on both basal-TMAP and nutrient agar medium displayed elementary branching mycelia fragmented into irregular rod and coccoid elements. Comparative 16S rDNA sequencing studies indicated that the strain A4 falls into the Rhodococcus erythropolis subclade and forms a monophyletic group with the type-strains of R. opacus, and R. wratislaviensis. Metabolites analysis by capillary electrophoresis, fast atom bombardment-mass spectrometry, and gas chromatography- mass spectrometry, showed trimethylamine (TMA) as the major metabolite beside β-alanine betaine and trimethylaminopropionaldehyde. Therefore, the possible degradation pathway of trimethylamino propanol in the isolated strain is through consequence oxidation of alcohol group (-OH) to aldehyde (-CHO) and acid (-COOH), and thereafter the cleavage of β-alanine betaine C-N bonds yielded trimethylamine and alkyl chain.

Recent Outbreaks of Highly Pathogenic Avian Influenza Virus in Chickens and Ducks in Egypt: Pathological Study

Clinically, chickens showed progressively developed clinical signs represented by sever cyanosis of the comb and wattles with hemorrhage at the shanks, depression, and ruffling feathers with loss of appetite and high daily mortalities. The ducks showed severe neurological signs as torticollus, seizures and inability to stand with mild signs of diarrhea and depression. Grossly, chickens showed hemorrhages and congestion in most of the organs particularly lung, liver, spleen, trachea and kidney. The examined ducks showed multiple petechial hemorrhages, multifocal hemorrhagic necrosis in the pancreas, pulmonary edema, congestion and hemorrhage in meninges and congestion in the skeletal muscles. Histopathology revealed severe congestion and hemorrhages in most of the organs particularly lung, liver and kidney. Microscopic erosive tracheitis, sever pulmonary congestion and perivascular oedema and lymphogranulocytic pneumonia were constant. The liver showed hepatocyts necrosis and lympho-granulocytic infiltration. The kidney showed renal tubular necrosis and diffuse congestion. Multifocal, neuronal necrosis, hemorrhages, multifocal glial nodules, lympho- histiocytic perivascular cuffing, and occasional neuronophagia were observed in the cerebrum. Other organs showed moderate changes.