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.

Observation of the Correlations between Pair Wise Interaction and Functional Organization of the Proteins, in the Protein Interaction Network of Saccaromyces Cerevisiae

Understanding the cell's large-scale organization is an interesting task in computational biology. Thus, protein-protein interactions can reveal important organization and function of the cell. Here, we investigated the correspondence between protein interactions and function for the yeast. We obtained the correlations among the set of proteins. Then these correlations are clustered using both the hierarchical and biclustering methods. The detailed analyses of proteins in each cluster were carried out by making use of their functional annotations. As a result, we found that some functional classes appear together in almost all biclusters. On the other hand, in hierarchical clustering, the dominancy of one functional class is observed. In brief, from interaction data to function, some correlated results are noticed about the relationship between interaction and function which might give clues about the organization of the proteins.

Genetic Polymorphism of Main Lactoproteins of Romanian Grey Steppe Breed in Preservation

The paper presents a part of the results obtained in a complex research project on Romanian Grey Steppe breed, owner of some remarkable qualities such as hardiness, longevity, adaptability, special resistance to ban weather and diseases and included in the genetic fund (G.D. no. 822/2008.) from Romania. Following the researches effectuated, we identified alleles of six loci, codifying the six types of major milk proteins: alpha-casein S1 (α S1-cz); beta-casein (β-cz); kappa-casein (K-cz); beta-lactoglobulin (β-lg); alpha-lactalbumin (α-la) and alpha-casein S2 (α S2-cz). In system αS1-cz allele αs1-Cn B has the highest frequency (0.700), in system β-cz allele β-Cn A2 ( 0.550 ), in system K-cz allele k-CnA2 ( 0.583 ) and heterozygote genotype AB ( 0.416 ) and BB (0.375), in system β-lg allele β-lgA1 has the highest frequency (0.542 ) and heterozygote genotype AB ( 0.500 ), in system α-la there is monomorphism for allele α-la B and similarly in system αS2-cz for allele αs2-Cn A. The milk analysis by the isoelectric focalization technique (I.E.F.) allowed the identification of a new allele for locus αS1-casein, for two of the individuals under analysis, namely allele called αS1-casein IRV. When experiments were repeated, we noticed that this is not a proteolysis band and it really was a new allele that has not been registered in the specialized literature so far. We identified two heterozygote individuals, carriers of this allele, namely: BIRV and CIRV. This discovery is extremely important if focus is laid on the national genetic patrimony.

Protein Production by Bacillus Subtilis Atcc 21332 in the Presence of Cymbopogon Essential Oils

Proteins levels produced by bacteria may be increased in stressful surroundings, such as in the presence of antibiotics. It appears that many antimicrobial agents or antibiotics, when used at low concentrations, have in common the ability to activate or repress gene transcription, which is distinct from their inhibitory effect. There have been comparatively few studies on the potential of antibiotics or natural compounds in nature as a specific chemical signal that can trigger a variety of biological functions. Therefore, this study was focusing on the effect of essential oils from Cymbopogon flexuosus and C. nardus in regulating proteins production by Bacillus subtilis ATCC 21332. The Minimum Inhibition Concentrations (MICs) of both essential oils on B. subtilis were determined by using microdilution assay, resulting 0.2% and 1.56% for each C. flexuosus and C. nardus subsequently. The bacteria were further exposed to each essential oils at concentration of 0.01XMIC for 2 days. The proteins were then isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Protein profile showed that a band with approximate size of 250 kD was appeared for the treated bacteria with essential oils. Thus, Bacillus subtilis ATCC 21332 in stressful condition with the presence of essential oils at low concentration could induce the protein production.

More Realistic Model for Simulating Min Protein Dynamics: Lattice Boltzmann Method Incorporating the Role of Nucleoids

The dynamics of Min proteins plays a center role in accurate cell division. Although the nucleoids may presumably play an important role in prokaryotic cell division, there is a lack of models to account for its participation. In this work, we apply the lattice Boltzmann method to investigate protein oscillation based on a mesoscopic model that takes into account the nucleoid-s role. We found that our numerical results are in reasonably good agreement with the previous experimental results On comparing with the other computational models without the presence of nucleoids, the highlight of our finding is that the local densities of MinD and MinE on the cytoplasmic membrane increases, especially along the cell width, when the size of the obstacle increases, leading to a more distinct cap-like structure at the poles. This feature indicated the realistic pattern and reflected the combination of Min protein dynamics and nucleoid-s role.

Baking Quality of Hulled Wheat Species in Organic Farming

The organic farmers use wider range of crop varieties than the conventional farming. Bread wheat is the most favorite and the most common food crop. The organic bread wheat is usually of worse technological quality. Therefore, it is supposed to be an attractive alternative to the hulled wheat species (einkorn, emmer wheat and spelt). Twenty-five hulled bread wheat varieties and control bread wheat ones were grown on the certified organic parcel in České Budějovice (the Czech Republic) between 2009 and 2012. Their baking quality was measured and evaluated with standard methods, and in accordance with ICC. The results have shown that the grain of hulled wheat varieties contain a lot of proteins in grains (up to 18 percent); even the organic hulled bread wheat varieties are characterized by such good baking quality. Einkorn and emmer wheat are of worse technological quality of proteins (low values of gluten index and Zeleny test), which is a disadvantage of these two wheat species. On the other hand, spelt wheat is of better technological quality and is similar to the control bread wheat varieties. Mixtures consisting of bread wheat, among others, are considered good alternatives; they may contribute to wider range of use of the hulled wheat species. It is one of the possibilities which may increase the proportion of proteins in bread wheat grains; the nutrition-rich hulled wheat grains may be also used in such way at the same time.

Analysis of DNA-Recognizing Enzyme Interaction using Deaminated Lesions

Deaminated lesions were produced via nitrosative oxidation of natural nucleobases; uracul (Ura, U) from cytosine (Cyt, C), hypoxanthine (Hyp, H) from adenine (Ade, A), and xanthine (Xan, X) and oxanine (Oxa, O) from guanine (Gua, G). Such damaged nucleobases may induce mutagenic problems, so that much attentions and efforts have been poured on the revealing of their mechanisms in vivo or in vitro. In this study, we employed these deaminated lesions as useful probes for analysis of DNA-binding/recognizing proteins or enzymes. Since the pyrimidine lesions such as Hyp, Oxa and Xan are employed as analogues of guanine, their comparative uses are informative for analyzing the role of Gua in DNA sequence in DNA-protein interaction. Several DNA oligomers containing such Hyp, Oxa or Xan substituted for Gua were designed to reveal the molecular interaction between DNA and protein. From this approach, we have got useful information to understand the molecular mechanisms of the DNA-recognizing enzymes, which have not ever been observed using conventional DNA oligomer composed of just natural nucleobases.

Improving Protein-Protein Interaction Prediction by Using Encoding Strategies and Random Indices

A New features are extracted and compared to improve the prediction of protein-protein interactions. The basic idea is to select and use the best set of features from the Tensor matrices that are produced by the frequency vectors of the protein sequences. Three set of features are compared, the first set is based on the indices that are the most common in the interacting proteins, the second set is based on the indices that tend to be common in the interacting and non-interacting proteins, and the third set is constructed by using random indices. Moreover, three encoding strategies are compared; that are based on the amino asides polarity, structure, and chemical properties. The experimental results indicate that the highest accuracy can be obtained by using random indices with chemical properties encoding strategy and support vector machine.

A Bayesian Kernel for the Prediction of Protein- Protein Interactions

Understanding proteins functions is a major goal in the post-genomic era. Proteins usually work in context of other proteins and rarely function alone. Therefore, it is highly relevant to study the interaction partners of a protein in order to understand its function. Machine learning techniques have been widely applied to predict protein-protein interactions. Kernel functions play an important role for a successful machine learning technique. Choosing the appropriate kernel function can lead to a better accuracy in a binary classifier such as the support vector machines. In this paper, we describe a Bayesian kernel for the support vector machine to predict protein-protein interactions. The use of Bayesian kernel can improve the classifier performance by incorporating the probability characteristic of the available experimental protein-protein interactions data that were compiled from different sources. In addition, the probabilistic output from the Bayesian kernel can assist biologists to conduct more research on the highly predicted interactions. The results show that the accuracy of the classifier has been improved using the Bayesian kernel compared to the standard SVM kernels. These results imply that protein-protein interaction can be predicted using Bayesian kernel with better accuracy compared to the standard SVM kernels.

Virulent-GO: Prediction of Virulent Proteins in Bacterial Pathogens Utilizing Gene Ontology Terms

Prediction of bacterial virulent protein sequences can give assistance to identification and characterization of novel virulence-associated factors and discover drug/vaccine targets against proteins indispensable to pathogenicity. Gene Ontology (GO) annotation which describes functions of genes and gene products as a controlled vocabulary of terms has been shown effectively for a variety of tasks such as gene expression study, GO annotation prediction, protein subcellular localization, etc. In this study, we propose a sequence-based method Virulent-GO by mining informative GO terms as features for predicting bacterial virulent proteins. Each protein in the datasets used by the existing method VirulentPred is annotated by using BLAST to obtain its homologies with known accession numbers for retrieving GO terms. After investigating various popular classifiers using the same five-fold cross-validation scheme, Virulent-GO using the single kind of GO term features with an accuracy of 82.5% is slightly better than VirulentPred with 81.8% using five kinds of sequence-based features. For the evaluation of independent test, Virulent-GO also yields better results (82.0%) than VirulentPred (80.7%). When evaluating single kind of feature with SVM, the GO term feature performs much well, compared with each of the five kinds of features.

Extraction and Characterisation of Protein Fraction from Date Palm Fruit Seeds

Date palm (Phoenix dactylifera L.) seeds are waste streams which are considered a major problem to the food industry. They contain potentially useful protein (10-15% of the whole date-s weight). Global production, industrialisation and utilisation of dates are increasing steadily. The worldwide production of date palm fruit has increased from 1.8 million tons in 1961 to 6.9 million tons in 2005, thus from the global production of dates are almost 800.000 tonnes of date palm seeds are not currently used [1]. The current study was carried out to convert the date palm seeds into useful protein powder. Compositional analysis showed that the seeds were rich in protein and fat 5.64 and 8.14% respectively. We used several laboratory scale methods to extract proteins from seed to produce a high protein powder. These methods included simple acid or alkali extraction, with or without ultrafiltration and phenol trichloroacetic acid with acetone precipitation (Ph/TCA method). The highest protein content powder (68%) was obtained by Ph/TCA method with yield of material (44%) whereas; the use of just alkali extraction gave the lowest protein content of 8%, and a yield of 32%.

Computational Identification of MicroRNAs and their Targets in two Species of Evergreen Spruce Tree (Picea)

MicroRNAs (miRNAs) are small, non-coding and regulatory RNAs about 20 to 24 nucleotides long. Their conserved nature among the various organisms makes them a good source of new miRNAs discovery by comparative genomics approach. The study resulted in 21 miRNAs of 20 pre-miRNAs belonging to 16 families (miR156, 157, 158, 164, 165, 168, 169, 172, 319, 390, 393, 394, 395, 400, 472 and 861) in evergreen spruce tree (Picea). The miRNA families; miR 157, 158, 164, 165, 168, 169, 319, 390, 393, 394, 400, 472 and 861 are reported for the first time in the Picea. All 20 miRNA precursors form stable minimum free energy stem-loop structure as their orthologues form in Arabidopsis and the mature miRNA reside in the stem portion of the stem loop structure. Sixteen (16) miRNAs are from Picea glauca and five (5) belong to Picea sitchensis. Their targets consist of transcription factors, growth related, stressed related and hypothetical proteins.

In Silico Analysis of Pax6 Interacting Proteins Indicates Missing Molecular Links in Development of Brain and Associated Disease

The PAX6, a transcription factor, is essential for the morphogenesis of the eyes, brain, pituitary and pancreatic islets. In rodents, the loss of Pax6 function leads to central nervous system defects, anophthalmia, and nasal hypoplasia. The haplo-insufficiency of Pax6 causes microphthalmia, aggression and other behavioral abnormalities. It is also required in brain patterning and neuronal plasticity. In human, heterozygous mutation of Pax6 causes loss of iris [aniridia], mental retardation and glucose intolerance. The 3- deletion in Pax6 leads to autism and aniridia. The phenotypes are variable in peneterance and expressivity. However, mechanism of function and interaction of PAX6 with other proteins during development and associated disease are not clear. It is intended to explore interactors of PAX6 to elucidated biology of PAX6 function in the tissues where it is expressed and also in the central regulatory pathway. This report describes In-silico approaches to explore interacting proteins of PAX6. The models show several possible proteins interacting with PAX6 like MITF, SIX3, SOX2, SOX3, IPO13, TRIM, and OGT. Since the Pax6 is a critical transcriptional regulator and master control gene of eye and brain development it might be interacting with other protein involved in morphogenesis [TGIF, TGF, Ras etc]. It is also presumed that matricelluar proteins [SPARC, thrombospondin-1 and osteonectin etc] are likely to interact during transport and processing of PAX6 and are somewhere its cascade. The proteins involved in cell survival and cell proliferation can also not be ignored.

SAF: A Substitution and Alignment Free Similarity Measure for Protein Sequences

The literature reports a large number of approaches for measuring the similarity between protein sequences. Most of these approaches estimate this similarity using alignment-based techniques that do not necessarily yield biologically plausible results, for two reasons. First, for the case of non-alignable (i.e., not yet definitively aligned and biologically approved) sequences such as multi-domain, circular permutation and tandem repeat protein sequences, alignment-based approaches do not succeed in producing biologically plausible results. This is due to the nature of the alignment, which is based on the matching of subsequences in equivalent positions, while non-alignable proteins often have similar and conserved domains in non-equivalent positions. Second, the alignment-based approaches lead to similarity measures that depend heavily on the parameters set by the user for the alignment (e.g., gap penalties and substitution matrices). For easily alignable protein sequences, it's possible to supply a suitable combination of input parameters that allows such an approach to yield biologically plausible results. However, for difficult-to-align protein sequences, supplying different combinations of input parameters yields different results. Such variable results create ambiguities and complicate the similarity measurement task. To overcome these drawbacks, this paper describes a novel and effective approach for measuring the similarity between protein sequences, called SAF for Substitution and Alignment Free. Without resorting either to the alignment of protein sequences or to substitution relations between amino acids, SAF is able to efficiently detect the significant subsequences that best represent the intrinsic properties of protein sequences, those underlying the chronological dependencies of structural features and biochemical activities of protein sequences. Moreover, by using a new efficient subsequence matching scheme, SAF more efficiently handles protein sequences that contain similar structural features with significant meaning in chronologically non-equivalent positions. To show the effectiveness of SAF, extensive experiments were performed on protein datasets from different databases, and the results were compared with those obtained by several mainstream algorithms.

Database Development and Discrimination Algorithms for Membrane Protein Functions

We have developed a database for membrane protein functions, which has more than 3000 experimental data on functionally important amino acid residues in membrane proteins along with sequence, structure and literature information. Further, we have proposed different methods for identifying membrane proteins based on their functions: (i) discrimination of membrane transport proteins from other globular and membrane proteins and classifying them into channels/pores, electrochemical and active transporters, and (ii) β-signal for the insertion of mitochondrial β-barrel outer membrane proteins and potential targets. Our method showed an accuracy of 82% in discriminating transport proteins and 68% to classify them into three different transporters. In addition, we have identified a motif for targeting β-signal and potential candidates for mitochondrial β-barrel membrane proteins. Our methods can be used as effective tools for genome-wide annotations.

Combining Gene and Chemo Therapy using Multifunctional Polymeric Micelles

Non-viral gene carriers composed of biodegradable polymers or lipids have been considered as a safer alternative for gene carriers over viral vectors. We have developed multi-functional nano-micelles for both drug and gene delivery application. Polyethyleneimine (PEI) was modified by grafting stearic acid (SA) and formulated to polymeric micelles (PEI-SA) with positive surface charge for gene and drug delivery. Our results showed that PEI-SA micelles provided high siRNA binding efficiency. In addition, siRNA delivered by PEI-SA carriers also demonstrated significantly high cellular uptake even in the presence of serum proteins. The post-transcriptional gene silencing efficiency was greatly improved by the polyplex formulated by 10k PEI-SA/siRNA. The amphiphilic structure of PEI-SA micelles provided advantages for multifunctional tasks; where the hydrophilic shell modified with cationic charges can electrostatically interact with DNA or siRNA, and the hydrophobic core can serve as payloads for hydrophobic drugs, making it a promising multifunctional vehicle for both genetic and chemotherapy application.

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.

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.

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.