A Heat-Inducible Transgene Expression System for Gene Therapy

Heat-inducible gene expression vectors are useful for hyperthermia-induced cancer gene therapy, because the combination of hyperthermia and gene therapy can considerably improve the therapeutic effects. In the present study, we developed an enhanced heat-inducible transgene expression system in which a heat-shock protein (HSP) promoter and tetracycline-responsive transactivator were combined. When the transactivator plasmid containing the tetracycline-responsive transactivator gene was co-transfected with the reporter gene expression plasmid, a high level of heat-induced gene expression was observed compared with that using the HSP promoter without the transactivator. In vitro evaluation of the therapeutic effect using HeLa cells showed that heat-induced therapeutic gene expression caused cell death in a high percentage of these cells, indicating that this strategy is promising for cancer gene therapy.

Effect of Wheat Flour Extraction Rates on Flour Composition, Farinographic Characteristics and Sensory Perception of Sourdough Naans

The effect of wheat flour extraction rates on flour composition, farinographic characteristics and the quality of sourdough naans was investigated. The results indicated that by increasing the extraction rate, the amount of protein, fiber, fat and ash increased, whereas moisture content decreased. Farinographic characteristic like water absorption and dough development time increased with an increase in flour extraction rate but the dough stabilities and tolerance indices were reduced with an increase in flour extraction rates. Titratable acidity for both sourdough and sourdough naans also increased along with flour extraction rate. The study showed that overall quality of sourdough naans were affected by both flour extraction rate and starter culture used. Sensory analysis of sourdough naans revealed that desirable extraction rate for sourdough naan was 76%.

Exact Pfaffian and N-Soliton Solutions to a (3+1)-Dimensional Generalized Integrable Nonlinear Partial Differential Equations

The objective of this paper is to use the Pfaffian technique to construct different classes of exact Pfaffian solutions and N-soliton solutions to some of the generalized integrable nonlinear partial differential equations in (3+1) dimensions. In this paper, I will show that the Pfaffian solutions to the nonlinear PDEs are nothing but Pfaffian identities. Solitons are among the most beneficial solutions for science and technology, from ocean waves to transmission of information through optical fibers or energy transport along protein molecules. The existence of multi-solitons, especially three-soliton solutions, is essential for information technology: it makes possible undisturbed simultaneous propagation of many pulses in both directions.

Optimization of Fiber Rich Gluten-Free Cookie Formulation by Response Surface Methodology

Most of the commercial gluten free products are nutritionally inferior when compared to gluten containing counterparts as manufacturers most often use the refined flours and starches. So it is possible that people on gluten free diet have low intake of fibre content. The foxtail millet flour and copra meal are gluten free and have high fibre and protein contents. The formulation of fibre rich gluten free cookies was optimized by response surface methodology considering independent process variables as proportion of Foxtail millet (Setaria italica) flour in mixed flour, fat content and guar gum. The sugar, sodium chloride, sodium bicarbonates and water were added in fixed proportion as 60, 1.0, 0.4 and 20% of mixed flour weight, respectively. Optimum formulation obtained for maximum spread ratio, fibre content, surface L-value, overall acceptability and minimum breaking strength were 80% foxtail millet flour in mixed flour, 42.8 % fat content and 0.05% guar gum.

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.

Detecting Community Structure in Amino Acid Interaction Networks

In this paper we introduce the notion of protein interaction network. This is a graph whose vertices are the protein-s amino acids and whose edges are the interactions between them. Using a graph theory approach, we observe that according to their structural roles, the nodes interact differently. By leading a community structure detection, we confirm this specific behavior and describe thecommunities composition to finally propose a new approach to fold a protein interaction network.

Effect of Stocking Density on Monosex Nile Tilapia Growth during Pond Culture in India

Stocking density is considered one of the important factors affecting fish growth. But, information related to impact of stocking density on growth performance of monosex tilapia population under the ecological conditions of Gangetic plains in West Bengal, India is limited. The aim of our study was to compare the growth potential of monosex tilapia at various stocking densities and to determine an ideal stocking density for culture of all-male monosex fish. The males were isolated by examination of genital papilla region and were stocked separately in 0.01 ha earthen ponds at different stocking densities (5000, 10000, 15000, 20000, 25000 and 30000 fingerlings/ha). It was found that the highest weight, length, daily weight gain, growth rate and protein content were observed for the 20000 fish/ha density class. Thus, culture of monosex tilapia at a density of 20000 fish/ha can be considered ideal for augmented production of the fish under Indian context.

Effect of Body Size and Condition Factor on Whole Body Composition of Hybrid (Catla catla ♂x Labeo rohita ♀) from Pakistan

In the present study, 49 Hybrid (Catla catla ♂ x Labeo rohita ♀) were sampled from Al-Raheem Fish Hatchery, Village Ali Pure Shamali, Jhang Road, 18 Km from Muzaffar Garh using a cast net and Live fishes were transported to research laboratory. Mean percentage for water found 79.13 %, ash 6.58 %, fat 2.22 % and protein content 12.06 % in whole wet body weight. It was observed that body constituents were found increasing in the same proportion with an increase in body weight while significant proportional increase was observed with total length. However, condition factor remained insignificant (P>0.05) with body constituents.

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.

Introducing Sequence-Order Constraint into Prediction of Protein Binding Sites with Automatically Extracted Templates

Search for a tertiary substructure that geometrically matches the 3D pattern of the binding site of a well-studied protein provides a solution to predict protein functions. In our previous work, a web server has been built to predict protein-ligand binding sites based on automatically extracted templates. However, a drawback of such templates is that the web server was prone to resulting in many false positive matches. In this study, we present a sequence-order constraint to reduce the false positive matches of using automatically extracted templates to predict protein-ligand binding sites. The binding site predictor comprises i) an automatically constructed template library and ii) a local structure alignment algorithm for querying the library. The sequence-order constraint is employed to identify the inconsistency between the local regions of the query protein and the templates. Experimental results reveal that the sequence-order constraint can largely reduce the false positive matches and is effective for template-based binding site prediction.

Sequence Relationships Similarity of Swine Influenza a (H1N1) Virus

In April 2009, a new variant of Influenza A virus subtype H1N1 emerged in Mexico and spread all over the world. The influenza has three subtypes in human (H1N1, H1N2 and H3N2) Types B and C influenza tend to be associated with local or regional epidemics. Preliminary genetic characterization of the influenza viruses has identified them as swine influenza A (H1N1) viruses. Nucleotide sequence analysis of the Haemagglutinin (HA) and Neuraminidase (NA) are similar to each other and the majority of their genes of swine influenza viruses, two genes coding for the neuraminidase (NA) and matrix (M) proteins are similar to corresponding genes of swine influenza. Sequence similarity between the 2009 A (H1N1) virus and its nearest relatives indicates that its gene segments have been circulating undetected for an extended period. Nucleic acid sequence Maximum Likelihood (MCL) and DNA Empirical base frequencies, Phylogenetic relationship amongst the HA genes of H1N1 virus isolated in Genbank having high nucleotide sequence homology. In this paper we used 16 HA nucleotide sequences from NCBI for computing sequence relationships similarity of swine influenza A virus using the following method MCL the result is 28%, 36.64% for Optimal tree with the sum of branch length, 35.62% for Interior branch phylogeny Neighber – Join Tree, 1.85% for the overall transition/transversion, and 8.28% for Overall mean distance.

Herpes Simplex Virus Type I Infection of Mice Testis and Effect on Fertility

The objective of current issue was to develop a model of testicular herpes simplex virus (HSV) type I infection for assessment of viral effect on fertility. 56 male mice were inoculated intraperitoneally with different concentrations of HSV on 8 day post partum. It was revealed that the optimal dose was 100 plaque forming units per mice as it provided testicular infection in 100% of survivors. HSV proteins were detected both in somatic and germ cells (spermatogonia, spermatocytes, spermatides). Although DNA load in testis was descending from 3 to 28 days post infection only 12.5% of infected males had offspring after mating with uninfected females comparing to 87.5% in control (p=0.012). These results are the first direct evidence for HSV impact in male sterility. Prepuberal mice appeared to be a suitable model for investigation of pathogenesis of virus-associated fertility disorders.

Detecting Remote Protein Evolutionary Relationships via String Scoring Method

The amount of the information being churned out by the field of biology has jumped manifold and now requires the extensive use of computer techniques for the management of this information. The predominance of biological information such as protein sequence similarity in the biological information sea is key information for detecting protein evolutionary relationship. Protein sequence similarity typically implies homology, which in turn may imply structural and functional similarities. In this work, we propose, a learning method for detecting remote protein homology. The proposed method uses a transformation that converts protein sequence into fixed-dimensional representative feature vectors. Each feature vector records the sensitivity of a protein sequence to a set of amino acids substrings generated from the protein sequences of interest. These features are then used in conjunction with support vector machines for the detection of the protein remote homology. The proposed method is tested and evaluated on two different benchmark protein datasets and it-s able to deliver improvements over most of the existing homology detection methods.

Dynamics of Blood Aminoacids in the Wounds- Treatment of Cows with Hydrocele Ointment

This article introduces the actual problem that is а study of proposed by the authors Hydrocele ointment in amino acids’ metabolism of cows’ blood in inflammation of traumatic origin. Hydrocele ointment has shown a positive effect on inflammatory process and amino acids’ metabolism of animals treated with the drug. Amino acid levels reached physiological parameters on the 10th day after treatment; in the control group this parameter was higher than normal.

Interaction Effect of DGAT1 and Composite Genotype of Beta-Kappa Casein on Economic Milk Production Traits in Crossbred Holstein

The objective was to determine the single gene and interaction effect of composite genotype of beta-kappa casein and DGAT1 gene on milk yield (MY) and milk composition, content of milk fat (%FAT), milk protein (%PRO), solid not fat (%SNF), and total solid (%TS) in crossbred Holstein cows. Two hundred and thirty- one cows were genotyped with PCR-RFLP for DGAT1 and composite genotype data of beta-kappa casein from previous work were used. Two model, (1), and (2), was used to estimate single gene effect, and interaction effect on the traits, respectively. The significance of interaction effects on all traits were detected. Most traits have consistent pattern of significant when model (1), and (2) were compared, except the effect of composite genotype of betakappa casein on %FAT, and the effect of DGAT1 on MY, which the significant difference was detected in only model (1).The results suggested that when the optimum of all traits was necessary, interaction effect should be concerned.

Production of H5N1 Hemagglutinin inTrichoplusia ni Larvae by a Novel Bi-cistronic Baculovirus Expression Vector

Highly pathogenic avian influenza (HPAI) H5N1 viruses have created demand for a cost-effective vaccine to prevent a pandemic of the disease. Here, we report that Trichoplusia ni (T. ni) larvae can act as a cost-effective bioreactor to produce recombinant HA5 (rH5HA) proteins as an potential effective vaccine for chickens. To facilitate the recombinant virus identification, virus titer determination and access the infected larvae, we employed the internal ribosome entry site (IRES) derived from Perina nuda virus (PnV, belongs to insect picorna like Iflavirus genus) to construct a bi-cistronic baculovirus expression vector that can express the rH5HA protein and enhanced green fluorescent protein (EGFP) simultaneously. Western blot analysis revealed that the 70 kDa rH5HA protein and partially cleaved products (40 kDa H5HA1) were generated in T. ni larvae infected with recombinant baculovirus carrying the H5HA gene. These data suggest that the baculovirus-larvae recombinant protein expression system could be a cost-effective platform for H5N1 vaccine production.

Computational Design of Inhibitory Agents of BMP-Noggin Interaction to Promote Osteogenesis

Bone growth factors, such as Bone Morphogenic Protein-2 (BMP-2) have been approved by the FDA to replace grafting for some surgical interventions, but the high dose requirement limits its use in patients. Noggin, an extracellular protein, blocks the effect of BMP-2 by binding to BMP. Preventing the BMP-2/noggin interaction will help increase the free concentration of BMP-2 and therefore should enhance its efficacy to induce bone formation. The work presented here involves computational design of novel small molecule inhibitory agents of BMP-2/noggin interaction, based on our current understanding of BMP-2, and its known putative ligands (receptors and antagonists). A successful acquisition of such an inhibitory agent of BMP-2/noggin interaction would allow clinicians to reduce the dose required of BMP-2 protein in clinical applications to promote osteogenesis. The available crystal structures of the BMPs, its receptors, and the binding partner noggin were analyzed to identify the critical residues involved in their interaction. In presenting this study, LUDI de novo design method was utilized to perform virtual screening of a large number of compounds from a commercially available library against the binding sites of noggin to identify the lead chemical compounds that could potentially block BMP-noggin interaction with a high specificity.

Sorting Primitives and Genome Rearrangementin Bioinformatics: A Unified Perspective

Bioinformatics and computational biology involve the use of techniques including applied mathematics, informatics, statistics, computer science, artificial intelligence, chemistry, and biochemistry to solve biological problems usually on the molecular level. Research in computational biology often overlaps with systems biology. Major research efforts in the field include sequence alignment, gene finding, genome assembly, protein structure alignment, protein structure prediction, prediction of gene expression and proteinprotein interactions, and the modeling of evolution. Various global rearrangements of permutations, such as reversals and transpositions,have recently become of interest because of their applications in computational molecular biology. A reversal is an operation that reverses the order of a substring of a permutation. A transposition is an operation that swaps two adjacent substrings of a permutation. The problem of determining the smallest number of reversals required to transform a given permutation into the identity permutation is called sorting by reversals. Similar problems can be defined for transpositions and other global rearrangements. In this work we perform a study about some genome rearrangement primitives. We show how a genome is modelled by a permutation, introduce some of the existing primitives and the lower and upper bounds on them. We then provide a comparison of the introduced primitives.

Effects of Feeding Glycerol to Lactating Dairy Cows on Milk Production and Composition

A study was conducted to determine the effect of feeding glycerol on dairy cows performance. Twenty four Holstein Friesian crossbred (>87.5% Holstein Friesian) lactating dairy cows in early lactation; averaging 13+2.4 kg of milk, 64+45 days in milk, 55+16 months old and 325+26 kg live weight, were stratified for milk yield, days in milk, age, stage of lactation and body weight, and then randomly allocated to three treatment groups. All cows were fed approximate 8 kg of concentrate together with ad libitum corn silage and freely access to clean water. Nil or 150 and 300g of glycerol were supplemented to the cows according to treatment groups. All cows consumed similar concentrate, corn silage and total DM and NELP. There were no significant differences in DM intake, CP intake, NELP intake, milk and milk composition yields. All cows had similar fat, protein, lactose, solid not fat and total solid percentage. All cows gain similar live weight. The present study indicated that, supplementation of glycerol did not enhance milk yield, milk composition and live weight change.

A Model to Study the Effect of Excess Buffers and Na+ Ions on Ca2+ Diffusion in Neuron Cell

Calcium is a vital second messenger used in signal transduction. Calcium controls secretion, cell movement, muscular contraction, cell differentiation, ciliary beating and so on. Two theories have been used to simplify the system of reaction-diffusion equations of calcium into a single equation. One is excess buffer approximation (EBA) which assumes that mobile buffer is present in excess and cannot be saturated. The other is rapid buffer approximation (RBA), which assumes that calcium binding to buffer is rapid compared to calcium diffusion rate. In the present work, attempt has been made to develop a model for calcium diffusion under excess buffer approximation in neuron cells. This model incorporates the effect of [Na+] influx on [Ca2+] diffusion,variable calcium and sodium sources, sodium-calcium exchange protein, Sarcolemmal Calcium ATPase pump, sodium and calcium channels. The proposed mathematical model leads to a system of partial differential equations which have been solved numerically using Forward Time Centered Space (FTCS) approach. The numerical results have been used to study the relationships among different types of parameters such as buffer concentration, association rate, calcium permeability.