Characterization and Evaluation of the Activity of Dipeptidyl Peptidase IV from the Black-Bellied Hornet Vespa basalis

Characterization and evaluation of the activity of Vespa basalis DPP-IV, which expressed in Spodoptera frugiperda 21 cells. The expression of rDPP-IV was confirmed by SDS–PAGE, Western blot analyses, LC-MS/MS and measurement of its peptidase specificity. One-step purification by Ni-NTA affinity chromatography and the total amount of rDPP-IV recovered was approximately 6.4mg per liter from infected culture medium; an equivalent amount would be produced by 1x109 infected Sf21 insect cells. Through the affinity purification led to highly stable rDPP-IV enzyme was recovered and with significant peptidase activity. The rDPP-IV exhibited classical Michaelis–Menten kinetics, with kcat/Km in the range of 10-500 mM-1×S-1 for the five synthetic substrates and optimum substrate is Ala-Pro-pNA. As expected in inhibition assay, the enzymatic activity of rDPP-IV was significantly reduced by 80 or 60% in the presence of sitagliptin (a DPP-IV inhibitor) or PMSF (a serine protease inhibitor), but was not apparently affected by iodoacetamide (a cysteine protease inhibitor).

Quantification of Peptides based on Isotope Dilution Surface Enhanced Raman Scattering

This study aims to demonstrate the quantification of peptides based on isotope dilution surface enhanced Raman scattering (IDSERS). SERS spectra of phenylalanine (Phe), leucine (Leu) and two peptide sequences TGQIFK (T13) and YSFLQNPQTSLCFSESIPTPSNR (T6) as part of the 22-kDa human growth hormone (hGH) were obtained on Ag-nanoparticle covered substrates. On the basis of the dominant Phe and Leu vibrational modes, precise partial least squares (PLS) prediction models were built enabling the determination of unknown T13 and T6 concentrations. Detection of hGH in its physiological concentration in order to investigate the possibility of protein quantification has been achieved.

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.

Ecosystems of Lake Sevan Basin-s Rivers in Armenia

Taking into account the importance of Lake Sevan and Lake Sevan basin-s rivers for Armenian economy, the main goals of our investigations were the documentation of water quality and the biodiversity of invertebrates developed in Lake Sevan basin-s rivers and selected tributaries. Moderately satisfied ecological condition for the biodiversity of Lake Sevan basin-s rivers has been established, and the changes in species- composition of zoobenthos in Lake Sevan were detected. A growing tendency of antibiotic resistance among E. coli isolates in water resources has been shown.

Therapeutic Product Preparation Bioprocess Modeling

An immunomodulator bioproduct is prepared in a batch bioprocess with a modified bacterium Pseudomonas aeruginosa. The bioprocess is performed in 100 L Bioengineering bioreactor with 42 L cultivation medium made of peptone, meat extract and sodium chloride. The optimal bioprocess parameters were determined: temperature – 37 0C, agitation speed - 300 rpm, aeration rate – 40 L/min, pressure – 0.5 bar, Dow Corning Antifoam M-max. 4 % of the medium volume, duration - 6 hours. This kind of bioprocesses are appreciated as difficult to control because their dynamic behavior is highly nonlinear and time varying. The aim of the paper is to present (by comparison) different models based on experimental data. The analysis criteria were modeling error and convergence rate. The estimated values and the modeling analysis were done by using the Table Curve 2D. The preliminary conclusions indicate Andrews-s model with a maximum specific growth rate of the bacterium in the range of 0.8 h-1.

A Report on Occurrence and Parasite-Host of Ligula intestinalis in Sattarkhan Lake(East Azerbaijan-Iran)

Ligula intestinalis is a three-host life-cycle Pseudophyllidean Cestode which in its plerocercoid stage infests a range of fresh water species. The objective of the present study was the worm occurrence within planctonic copepods, fishes and piscivorous birds and examine of parasite-hosts samples in the Lake of Sattarkhan Dam (near the city of Ahar, East Azerbaijan, Iran). Fish sample were collected with fyke and gill nets and the abdominal cavity was examined for the presence of ligula. Zooplanktons were captured using a planktonic net and occurrence of parasitic larval form in the body cavity was determined. Piscivorous birds were selected by telescope, they hunted and dissected for presence of parasite eggs in their gut. Results indicated that prevalence of infection was 16% for cyclopid copepoda and majority of infected cyclopid were female Cyclops. Investigation of 310 fishes specimens were indicated to infection of five species of cyprinid fishes. In addition, results indicated to manipulation of six species of migratory aquatic and semi aquatic birds by ligula. Obtained results are in agreement by previous studies. Its definite in this study that all of fishes in Sattarkhan Lake capable to infection, its important for health because they capture by native people and it is documented that ligula can be introduce as a zoonose. It's seemed that to prevent from disperses of parasite and restricted of infection, biological elimination can be effective and it's necessary to inform native people about sanitation.

Hydrolysis of Hull-Less Pumpkin Oil Cake Protein Isolate by Pepsin

The present work represents an investigation of the hydrolysis of hull-less pumpkin (Cucurbita Pepo L.) oil cake protein isolate (PuOC PI) by pepsin. To examine the effectiveness and suitability of pepsin towards PuOC PI the kinetic parameters for pepsin on PuOC PI were determined and then, the hydrolysis process was studied using Response Surface Methodology (RSM). The hydrolysis was carried out at temperature of 30°C and pH 3.00. Time and initial enzyme/substrate ratio (E/S) at three levels were selected as the independent parameters. The degree of hydrolysis, DH, was mesuared after 20, 30 and 40 minutes, at initial E/S of 0.7, 1 and 1.3 mA/mg proteins. Since the proposed second-order polynomial model showed good fit with the experimental data (R2 = 0.9822), the obtained mathematical model could be used for monitoring the hydrolysis of PuOC PI by pepsin, under studied experimental conditions, varying the time and initial E/S. To achieve the highest value of DH (39.13 %), the obtained optimum conditions for time and initial E/S were 30 min and 1.024 mA/mg proteins.

Biological Effects of a Carbohydrate-Binding Protein from an Annelid, Perinereis nuntia Against Human and Phytopathogenic Microorganisms

Lectins have a good scope in current clinical microbiology research. In the present study evaluated the antimicrobial activities of a D-galactose binding lectin (PnL) was purified from the annelid, Perinereis nuntia (polychaeta) by affinity chromatography. The molecular mass of the lectin was determined to be 32 kDa as a single polypeptide by SDS-PAGE under both reducing and non-reducing conditions. The hemagglutinating activity of the PnL showed against trypsinized and glutaraldehyde-fixed human erythrocytes was specifically inhibited by D-Gal, GalNAc, Galβ1-4Glc and Galα1-6Glc. PnL was evaluated for in vitro antibacterial screening studies against 11 gram-positive and gram-negative microorganisms. From the screening results, it was revealed that PnL exhibited significant antibacterial activity against gram-positive bacteria. Bacillus megaterium showed the highest growth inhibition by the lectin (250 μg/disc). However, PnL did not inhibit the growth of gram-negative bacteria such as Vibrio cholerae and Pseudomonas sp. PnL was also examined for in vitro antifungal activity against six fungal phytopathogens. PnL (100 μg/mL) inhibited the mycelial growth of Alternaria alternata (24.4%). These results indicate that future findings of lectin applications obtained from annelids may be of importance to life sciences.

Extended Study on Removing Gaussian Noise in Mechanical Engineering Drawing Images using Median Filters

In this paper, an extended study is performed on the effect of different factors on the quality of vector data based on a previous study. In the noise factor, one kind of noise that appears in document images namely Gaussian noise is studied while the previous study involved only salt-and-pepper noise. High and low levels of noise are studied. For the noise cleaning methods, algorithms that were not covered in the previous study are used namely Median filters and its variants. For the vectorization factor, one of the best available commercial raster to vector software namely VPstudio is used to convert raster images into vector format. The performance of line detection will be judged based on objective performance evaluation method. The output of the performance evaluation is then analyzed statistically to highlight the factors that affect vector quality.

Molecular Mechanism of Amino Acid Discrimination for the Editing Reaction of E.coli Leucyl-tRNA Synthetase

Certain tRNA synthetases have developed highly accurate molecular machinery to discriminate their cognate amino acids. Those aaRSs achieve their goal via editing reaction in the Connective Polypeptide 1 (CP1). Recently mutagenesis studies have revealed the critical importance of residues in the CP1 domain for editing activity and X-ray structures have shown binding mode of noncognate amino acids in the editing domain. To pursue molecular mechanism for amino acid discrimination, molecular modeling studies were performed. Our results suggest that aaRS bind the noncognate amino acid more tightly than the cognate one. Finally, by comparing binding conformations of the amino acids in three systems, the amino acid binding mode was elucidated and a discrimination mechanism proposed. The results strongly reveal that the conserved threonines are responsible for amino acid discrimination. This is achieved through side chain interactions between T252 and T247/T248 as well as between those threonines and the incoming amino acids.

Italians- Social and Emotional Loneliness: The Results of Five Studies

Subjective loneliness describes people who feel a disagreeable or unacceptable lack of meaningful social relationships, both at the quantitative and qualitative level. The studies to be presented tested an Italian 18-items self-report loneliness measure, that included items adapted from scales previously developed, namely a short version of the UCLA (Russell, Peplau and Cutrona, 1980), and the 11-items Loneliness scale by De Jong-Gierveld & Kamphuis (JGLS; 1985). The studies aimed at testing the developed scale and at verifying whether loneliness is better conceptualized as a unidimensional (so-called 'general loneliness') or a bidimensional construct, namely comprising the distinct facets of social and emotional loneliness. The loneliness questionnaire included 2 singleitem criterion measures of sad mood, and social contact, and asked participants to supply information on a number of socio-demographic variables. Factorial analyses of responses obtained in two preliminary studies, with 59 and 143 Italian participants respectively, showed good factor loadings and subscale reliability and confirmed that perceived loneliness has clearly two components, a social and an emotional one, the latter measured by two subscales, a 7-item 'general' loneliness subscale derived from UCLA, and a 6–item 'emotional' scale included in the JGLS. Results further showed that type and amount of loneliness are related, negatively, to frequency of social contacts, and, positively, to sad mood. In a third study data were obtained from a nation-wide sample of 9.097 Italian subjects, 12 to about 70 year-olds, who filled the test on-line, on the Italian web site of a large-audience magazine, Focus. The results again confirmed the reliability of the component subscales, namely social, emotional, and 'general' loneliness, and showed that they were highly correlated with each other, especially the latter two. Loneliness scores were significantly predicted by sex, age, education level, sad mood and social contact, and, less so, by other variables – e.g., geographical area and profession. The scale validity was confirmed by the results of a fourth study, with elderly men and women (N 105) living at home or in residential care units. The three subscales were significantly related, among others, to depression, and to various measures of the extension of, and satisfaction with, social contacts with relatives and friends. Finally, a fifth study with 315 career-starters showed that social and emotional loneliness correlate with life satisfaction, and with measures of emotional intelligence. Altogether the results showed a good validity and reliability in the tested samples of the entire scale, and of its components.

Effect of Crude Extract from Bacillus Subtilis LB5 Cultivated Broth on Conidial Germination of Colletotrichum Gloeosporioides

Bacillus subtilis strain LB5 produced lipopeptide antibiotic iturin A-2 in liquid medium. Crude extract from cell-free supernatant of B. subtilis cultivated broth extracted with n-butanol showed antifungal activity to conidial germination of Colletotrichum gloeosporioides. The germination of conidia was completely inhibited by crude extract. The ultrastructure of conidia after treated with crude extract was found an accumulation of vesiclelike material between cell wall and plasma membrane while this accumulation was not observed in untreated and germinated conidia. Besides, the cell wall was not affected by crude extract.

Robust Statistics Based Algorithm to Remove Salt and Pepper Noise in Images

In this paper, a robust statistics based filter to remove salt and pepper noise in digital images is presented. The function of the algorithm is to detect the corrupted pixels first since the impulse noise only affect certain pixels in the image and the remaining pixels are uncorrupted. The corrupted pixels are replaced by an estimated value using the proposed robust statistics based filter. The proposed method perform well in removing low to medium density impulse noise with detail preservation upto a noise density of 70% compared to standard median filter, weighted median filter, recursive weighted median filter, progressive switching median filter, signal dependent rank ordered mean filter, adaptive median filter and recently proposed decision based algorithm. The visual and quantitative results show the proposed algorithm outperforms in restoring the original image with superior preservation of edges and better suppression of impulse noise

Mass Casualty Acute Pepper Spray Inhalation Respiratory Effect Severity

Pepper spray use has gained momentum since 1992 and although the active ingredient is readily available, it is considered a weapon with restricted use in many regions, including The Bahamas. In light of controversy in the literature regarding the severity of presenting respiration complaints among individuals postacute exposure of pepper spray inhalation, this descriptive case series study was conducted to assess the respiratory status of persons evaluated during a mass casualty in The Bahamas. Parameters noted were patients- demographics and respiration severity determined via clinical examination findings, disposition and follow-up review of the 20 persons. Their most common complaint was difficulty breathing post exposure. Two required admission and stayed for

In Vitro Antibacterial and Antifungal Effects of a 30 kDa D-Galactoside-Specific Lectin from the Demosponge, Halichondria okadai

The present study has been taken to explore the screening of in vitro antimicrobial activities of D-galactose-binding sponge lectin (HOL-30). HOL-30 was purified from the marine demosponge Halichondria okadai by affinity chromatography. The molecular mass of the lectin was determined to be 30 kDa with a single polypeptide by SDS-PAGE under non-reducing and reducing conditions. HOL-30 agglutinated trypsinized and glutaraldehydefixed rabbit and human erythrocytes with preference for type O erythrocytes. The lectin was subjected to evaluation for inhibition of microbial growth by the disc diffusion method against eleven human pathogenic gram-positive and gram-negative bacteria. The lectin exhibited strong antibacterial activity against gram-positive bacteria, such as Bacillus megaterium and Bacillus subtilis. However, it did not affect against gram-negative bacteria such as Salmonella typhi and Escherichia coli. The largest zone of inhibition was recorded of Bacillus megaterium (12 in diameter) and Bacillus subtilis (10 mm in diameter) at a concentration of the lectin (250 μg/disc). On the other hand, the antifungal activity of the lectin was investigated against six phytopathogenic fungi based on food poisoning technique. The lectin has shown maximum inhibition (22.83%) of mycelial growth of Botrydiplodia theobromae at a concentration of 100 μg/mL media. These findings indicate that the lectin may be of importance to clinical microbiology and have therapeutic applications.

Adjustment of a PET Scanner for PEPT

Positron emission particle tracking (PEPT) is a technique in which a single radioactive tracer particle can be accurately tracked as it moves. A limitation of PET is that in order to reconstruct a tomographic image it is necessary to acquire a large volume of data (millions of events), so it is difficult to study rapidly changing systems. By considering this fact, PEPT is a very fast process compared with PET. In PEPT detecting both photons defines a line and the annihilation is assumed to have occurred somewhere along this line. The location of the tracer can be determined to within a few mm from coincident detection of a small number of pairs of back-to-back gamma rays and using triangulation. This can be achieved many times per second and the track of a moving particle can be reliably followed. This technique was invented at the University of Birmingham [1]. The attempt in PEPT is not to form an image of the tracer particle but simply to determine its location with time. If this tracer is followed for a long enough period within a closed, circulating system it explores all possible types of motion. The application of PEPT to industrial process systems carried out at the University of Birmingham is categorized in two subjects: the behaviour of granular materials and viscous fluids. Granular materials are processed in industry for example in the manufacture of pharmaceuticals, ceramics, food, polymers and PEPT has been used in a number of ways to study the behaviour of these systems [2]. PEPT allows the possibility of tracking a single particle within the bed [3]. Also PEPT has been used for studying systems such as: fluid flow, viscous fluids in mixers [4], using a neutrally-buoyant tracer particle [5].

Depression and Anxiety Levels in Armenian Crohn's Disease Patients

The Zung self-depression scale and Beck Anxiety Inventory were used to study the depression and anxiety levels of Armenian Crohn's disease patients, as well as to reveal the relation between emotional status and placebo effect of these patients. Despite of registered high levels of depression and anxiety, the high placebo rate during investigations was described. The importance of use of psychotherapies for optimal outcomes during treatments of Crohn's disease is obvious.

Noise Reduction in Image Sequences using an Effective Fuzzy Algorithm

In this paper, we propose a novel spatiotemporal fuzzy based algorithm for noise filtering of image sequences. Our proposed algorithm uses adaptive weights based on a triangular membership functions. In this algorithm median filter is used to suppress noise. Experimental results show when the images are corrupted by highdensity Salt and Pepper noise, our fuzzy based algorithm for noise filtering of image sequences, are much more effective in suppressing noise and preserving edges than the previously reported algorithms such as [1-7]. Indeed, assigned weights to noisy pixels are very adaptive so that they well make use of correlation of pixels. On the other hand, the motion estimation methods are erroneous and in highdensity noise they may degrade the filter performance. Therefore, our proposed fuzzy algorithm doesn-t need any estimation of motion trajectory. The proposed algorithm admissibly removes noise without having any knowledge of Salt and Pepper noise density.

Toward Full Public E-Service Environment in Developing Countries

Changing technology and increased constituent demand for government services derive the need for governmental responsiveness. The government organisations in the developing countries will be under increased pressure to change their bureaucratic systems to be able to respond rapidly to changing and increasing requirements and rapid technology advancements. This paper aims to present a conceptual framework for explaining the main barriers and drivers of public e-service development. Therefore, the framework provides a basic context within which the process and practice of E-Service can be implemented successfully in the public sector organisations. The framework is flexible enough to be adopted by governments at different levels; national or local by developing countries around the world.

Heat and Mass Transfer in a Solar Dryer with Biomass Backup Burner

Majority of pepper farmers in Malaysia are using the open-sun method for drying the pepper berries. This method is time consuming and exposed the berries to rain and contamination. A maintenance-friendly and properly enclosed dryer is therefore desired. A dryer design with a solar collector and a chimney was studied and adapted to suit the needs of small-scale pepper farmers in Malaysia. The dryer will provide an environment with an optimum operating temperature meant for drying pepper berries. The dryer model was evaluated by using commercially available computational fluid dynamic (CFD) software in order to understand the heat and mass transfer inside the dryer. Natural convection was the only mode of heat transportation considered in this study as in accordance to the idea of having a simple and maintenance-friendly design. To accommodate the effect of low buoyancy found in natural convection driers, a biomass burner was integrated into the solar dryer design.