Publications | Biotechnology and Bioengineering
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 276

World Academy of Science, Engineering and Technology

[Biotechnology and Bioengineering]

Online ISSN : 1307-6892

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

Authors: Muhammad Y.K. Barozai, Ifthikhar A. Baloch, M. Din

Abstract:

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.

Keywords: BLAST, Comparative Genomics, Micro-RNAs, Spruce

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35 An Evaluation of Pesticide Stress Induced Proteins in three Cyanobacterial Species-Anabaena Fertilissima, Aulosira Fertilissima and Westiellopsis Prolifica using SDS-PAGE

Authors: Nirmal Kumar, Rita N. Kumar, Anubhuti Bora, Manmeet Kaur Amb

Abstract:

The whole-cell protein-profiling technique was evaluated for studying differences in banding pattern of three different species of Cyanobacteria i.e. Anabaena fertilissima, Aulosira fertilissima and Westiellopsis prolifica under the influence of four different pesticides-2,4-D (Ethyl Ester of 2,4-Dichloro Phenoxy Acetic Acid), Pencycuron (N-[(4-chlorophenyl)methyl]-Ncyclopentyl- N'–phenylurea), Endosulfan (6,7,8,9,10,10hexachloro- 1,5,5a,6,9,9a-hexahydro-6,9-methano-2,4,3-benzodioxathiepine-3- oxide) and Tebuconazole (1-(4-Chlorophenyl)-4,4-dimethyl-3-(1,2,4- triazol-1-ylmethyl)pentan-3-ol). Whole-cell extracts were obtained by sonication treatment (Sonifier cell disruptor -Branson Digital Sonifier S-450D, USA) and were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). SDS-PAGE analyses of the total protein profile of Anabaena fertilissima, Aulosira fertilissima and Westiellopsis prolifica showed a linear decrease in the protein content with increasing pesticide stress when administered to different concentrations of 2, 4-D, Pencycuron, Endosulfan and Tebuconazole. The results indicate that different stressors exert specific effects on cyanobacterial protein synthesis.

Keywords: Cyanobacteria, pesticide, SDS-PAGE

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34 Differential Sensitivity of Nitrogen-Fixing, Filamentous Cyanobacterial Species to an Organochlorine Insecticide - 6, 7, 8, 9, 10, 10- Hexachloro-1, 5, 5a, 6, 9, 9a-Hexahydro-6, 9- Methano-2, 4, 3-Benzodioxathiepine-3-Oxide

Authors: Nirmal J.I. Kumar, Anubhuti A. Bora, Manmeet K. Amb

Abstract:

Application of pesticides in the paddy fields has deleterious effects on non-target organisms including cyanobacteria which are photosynthesizing and nitrogen fixing micro-organisms contributing significantly towards soil fertility and crop yield. Pesticide contamination in the paddy fields has manifested into a serious global environmental concern. To study the effect of one such pesticide, three cyanobacterial strains; Anabaena fertilissima, Aulosira fertilissima and Westiellopsis prolifica were selected for their stress responses to an Organochlorine insecticide - 6, 7, 8, 9, 10, 10-hexachloro-1, 5, 5a, 6, 9, 9a-hexahydro-6, 9-methano-2, 4, 3- benzodioxathiepine-3-oxide, with reference to their photosynthesic pigments-chlorophyll-a and carotenoids as well as accessory pigments-phycobiliproteins (phycocyanin, allophycocyanin and phycoerythrin), stress induced biochemical metabolites like carbohydrates, proteins, amino acids, phenols and enzymes-nitrate reductase, glutamine synthetase and succinate dehydrogenase. All the three cyanobacterial strains were adversely affected by the insecticide doses and inhibition was dose dependent. Reduction in photosynthetic and accessory pigments, metabolites, nitrogen fixing and respiratory enzymes of the test organisms were accompanied with an initial increase in their total protein at lower Organochlorine doses. On the other hand, increased amount of phenols in all the insecticide treated concentrations was indicative of stressed activities of the organisms.

Keywords: biochemical metabolites, endosulfan, enzymes, pigments

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33 Effects of a Nectandra Membranacea Extract on Labeling of Blood Constituents with Technetium-99m and on the Morphology of Red Blood Cells

Authors: Silvana R.F. Moreno, Jorge J. Carvalho, Ana L. Nascimento, Mario Pereira, Luiz Q. A. Caldas, Mário Bernardo-Filho

Abstract:

The aim of this in vitro study was to evaluate the possible interference of a Nectandra membranacea extract (i) on the labeling of blood cells (BC), (ii) on the labeling process of BC and plasma (P) proteins with technetium-99m (Tc-99m) and (iii) on the morphology of red blood cells (RBC). Blood samples were incubated with a Nectandra membranacea crude extract, stannous chloride, Tc- 99m (sodium pertechnetate) was added, and soluble (SF) and insoluble (IF) fractions were isolated. Morphometry studies were performed with blood samples incubated with Nectandra membranacea extract. The results show that the Nectandra membranacea extract does not promote significant alteration of the labeling of BC, IF-P and IF-BC. The Nectandra membranacea extract was able to alter the erythrocyte membrane morphology, but these morphological changes were not capable to interfere on the labeling of blood constituents with Tc-99m.

Keywords: in vitro study, Nectandra membranacea, red bloodcell, technetium-99m

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

Authors: Essam Al-Daoud

Abstract:

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.

Keywords: protein-protein interactions, random indices, encoding strategies, support vector machine.

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31 A Pairwise-Gaussian-Merging Approach: Towards Genome Segmentation for Copy Number Analysis

Authors: Chih-Hao Chen, Hsing-Chung Lee, Qingdong Ling, Hsiao-Jung Chen, Sun-Chong Wang, Li-Ching Wu, H.C. Lee

Abstract:

Segmentation, filtering out of measurement errors and identification of breakpoints are integral parts of any analysis of microarray data for the detection of copy number variation (CNV). Existing algorithms designed for these tasks have had some successes in the past, but they tend to be O(N2) in either computation time or memory requirement, or both, and the rapid advance of microarray resolution has practically rendered such algorithms useless. Here we propose an algorithm, SAD, that is much faster and much less thirsty for memory – O(N) in both computation time and memory requirement -- and offers higher accuracy. The two key ingredients of SAD are the fundamental assumption in statistics that measurement errors are normally distributed and the mathematical relation that the product of two Gaussians is another Gaussian (function). We have produced a computer program for analyzing CNV based on SAD. In addition to being fast and small it offers two important features: quantitative statistics for predictions and, with only two user-decided parameters, ease of use. Its speed shows little dependence on genomic profile. Running on an average modern computer, it completes CNV analyses for a 262 thousand-probe array in ~1 second and a 1.8 million-probe array in 9 seconds

Keywords: Cancer, pathogenesis, chromosomal aberration, copy number variation, segmentation analysis.

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30 Temperature-dependent Structural Perturbation of Tuna Myoglobin

Authors: Yoshihiro Ochiai

Abstract:

To unveil the mechanism of fast autooxidation of fish myoglobins, the effect of temperature on the structural change of tuna myoglobin was investigated. Purified myoglobin was subjected to preincubation at 5, 20, 50 and 40oC. Overall helical structural decay through thermal treatment up to 95oC was monitored by circular dichroism spectrometry, while the structural changes around the heme pocket was measured by ultraviolet/visible absorption spectrophotometry. As a result, no essential structural change of myoglobin was observed under 30oC, roughly equivalent to their body temperature, but the structure was clearly damaged at 40oC. The Soret band absorption hardly differed irrespective of preincubation temperature, suggesting that the structure around the heme pocket was not perturbed even after thermal treatment.

Keywords: denaturation, myoglobin, stability, tuna.

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29 Intrinsic Electromagnetic Fields and Atom-Field Coupling in Living Cells

Authors: Masroor H. S. Bukhari, Z. H. Shah

Abstract:

The possibility of intrinsic electromagnetic fields within living cells and their resonant self-interaction and interaction with ambient electromagnetic fields is suggested on the basis of a theoretical and experimental study. It is reported that intrinsic electromagnetic fields are produced in the form of radio-frequency and infra-red photons within atoms (which may be coupled or uncoupled) in cellular structures, such as the cell cytoskeleton and plasma membrane. A model is presented for the interaction of these photons among themselves or with atoms under a dipole-dipole coupling, induced by single-photon or two-photon processes. This resonance is manifested by conspicuous field amplification and it is argued that it is possible for these resonant photons to undergo tunnelling in the form of evanescent waves to a short range (of a few nanometers to micrometres). This effect, suggested as a resonant photon tunnelling mechanism in this report, may enable these fields to act as intracellular signal communication devices and as bridges between macromolecules or cellular structures in the cell cytoskeleton, organelles or membrane. A brief overview of an experimental technique and a review of some preliminary results are presented, in the detection of these fields produced in living cell membranes under physiological conditions.

Keywords: bioelectromagnetism, cell membrane, evanescentwaves, photon tunnelling, resonance

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28 Determination of Alkaline Protease Production In Serratia Marcescens Sp7 Using Agro Wastes As Substrate Medium, Optimization Of Production Parameters And Purification Of The Enzyme

Authors: Baby Joseph, Sankarganesh Palaniyandi

Abstract:

The enzyme alkaline protease production was determined under solid state fermentation using the soil bacteria Serratia marcescens sp7. The maximum production was obtained from wheat bran medium than ground nut shell and chemically defined medium. The physiological fermentation factors such as pH of the medium (pH 8), Temperature (40oC) and incubation time (48 hrs) played a vital role in alkaline protease production in all the above. 100Mm NaCl has given better resolution during elution of the enzymes. The enzyme production was found to be associated with growth of the bacterial culture.

Keywords: Alkaline protease, Wheat bran, Ground nut shell, Serratia marcescens

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27 Hair Mechanical Properties Depending on Age and Origin

Authors: Meriem Benzarti, Mohamed Ben Tkaya, Cyril Pailler Mattei, Hassan Zahouani

Abstract:

Hair is a non homogenous complex material which can be associated with a polymer. It is made up 95% of Keratin. Hair has a great social significance for human beings. In the High Middle Ages, for example, long hairs have been reserved for kings and nobles. Most common interest in hair is focused on hair growth, hair types and hair care, but hair is also an important biomaterial which can vary depending on ethnic origin or on age, hair colour for example can be a sign of ethnic ancestry or age (dark hair for Asiatic, blond hair for Caucasian and white hair for old people in general). In this context, different approaches have been conducted to determine the differences in mechanical properties and characterize the fracture topography at the surface of hair depending on its type and its age. A tensile testing machine was especially designed to achieve tensile tests on hair. This device is composed of a microdisplacement system and a force sensor whose peak load is limited to 3N. The curves and the values extracted from each experiment, allow us to compare the evolution of the mechanical properties from one hair to another. Observations with a Scanning Electron Microscope (SEM) and with an interferometer were made on different hairs. Thus, it is possible to access the cuticle state and the fracture topography for each category.

Keywords: Hair, relaxation test, SEM, interferometer, mechanical properties.

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26 In vitro Study of Antibacterial Activity of Cymbopogon citratus

Authors: C.K. Hindumathy

Abstract:

Alcohol and water extracts of Cymbopogon citratus was investigated for anti-bacterial properties and phytochemical constituents. The extract was screened against four gram-negative bacteria Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus vulgaris) and two grampositive bacteria Bacillus subtilis and Staphylococcus aureus at four different concentrations (1:1, 1:5, 1:10 and 1:20) using disc diffusion method. The antibacterial examination was by disc diffusion techniques, while the photochemical constituents were investigated using standard chemical methods. Results showed that the extracts inhibited the growth of standard and local strains of the organisms used. The treatments were significantly different (P = 0.05). The minimum inhibitory concentration of the extracts against the tested microorganisms ranged between 150mg/ml and 50mg/ml. The alcohol extracts were found to be generally more effective than the water extract. The photochemical analysis revealed the presence of alkaloids and phenol but absence of cardiac and cyanogenic glycosides. The presence of alkaloid and phenols were inferred as being responsible for the anti-bacterial properties of the extracts.

Keywords: Cymbopogon citratus; gram negative and grampositive

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25 The First Prevalence Report of Direct Identification and Differentiation of B. abortus and B. melitensis using Real Time PCR in House Mouse of Iran

Authors: A. Doosti, S. Moshkelani

Abstract:

Brucellosis is a zoonotic disease; its symptoms and appearances are not exclusive in human and its traditional diagnosis is based on culture, serological methods and conventional PCR. For more sensitive, specific detection and differentiation of Brucella spp., the real time PCR method is recommended. This research has performed to determine the presence and prevalence of Brucella spp. and differentiation of Brucella abortus and Brucella melitensis in house mouse (Mus musculus) in west of Iran. A TaqMan analysis and single-step PCR was carried out in total 326 DNA of Mouse's spleen samples. From the total number of 326 samples, 128 (39.27%) gave positive results for Brucella spp. by conventional PCR, also 65 and 32 out of the 128 specimens were positive for B. melitensis, B. abortus, respectively. These results indicate a high presence of this pathogen in this area and that real time PCR is considerably faster than current standard methods for identification and differentiation of Brucella species. To our knowledge, this study is the first prevalence report of direct identification and differentiation of B. abortus and B. melitensis by real time PCR in mouse tissue samples in Iran.

Keywords: Differentiation, B. abortus, B. melitensis, TaqManprobe, Iran.

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24 Differentiation between Common Tick Species Using Molecular Biology Techniques in Saudi Arabia

Authors: Kholoud A. Al-Shammery , Badr El-Sabah A. Fetoh, Ahmed M. Alshammari

Abstract:

Protein and Esterase electrophoresis were used to genetically identify two Saudi tick species. Engorged females of the camel tick Hyalomma dromedarii (Koch) (Acari: Ixodidae) and the cattle tick Boophilus annulatus (Say) (Acari: Ixodidae) ticks collected from infested camels and cattle in the animals resting house at Hail region in KSA were used. The results showed that there are a variation in both of protein and esterase activity levels and a high polymorphism within and between the genera and species of Hyalomma and Boophilus . In conclusion, the protein and esterase electrophoretic analysis used in the present study could successfully distinguish among tick species, commonly found in Saudi Arabia.

Keywords: Molecular biology, The camel tick Hyalomma dromedarii, The cattle tick Boophilus annulatus , Ticks.

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23 Regulatory Effects of Carbon Sources on Tabtoxin Production (A β-lactam Phytotoxin of Pseudomonas syringae pv. tabaci)

Authors: N. Messaadia, D. Harzallah

Abstract:

The effects of divers carbon substrates were investigated for the tabtoxin production of an isolated pathogenic Pseudomonas syringae pv. tabaci, the causal agent of wildfire of tobacco and are discussed in relation to the bacterium growth. The isolated organism was grown in batch culture on Woolley's medium (28°C, 200 rpm, during 5 days). The growth has been measured by the optical density (OD) at 620 nm and the tabtoxin production quantified by Escherichia coli (K-12) bioassay technique. The growth and the tabtoxin production were both influenced by the substrates (sugars, amino acids, organic acids) used, each, as a sole carbon source and as a supplement for the same amino acids. The most significant quantities of tabtoxin were obtained in presence of some amino acids used as sole carbon source and/or as supplement.

Keywords: Amino acid supplement, carbon substrates, batch culture, Pseudomonas syringae pv. tabaci.

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22 Prevalence of Epstein-Barr Virus Latent Membrane Protein-1 in Jordanian Patients with Hodgkin's Lymphoma and Non- Hodgkin's Lymphoma

Authors: Fawzi Irshaid, Adnan Jaran, Fatiha Dilmi, Khaled Tarawneh, Raji Hadeth, Ahad Al-Khatib

Abstract:

The aim of this study was to estimate the frequency of EBV infection in Hodgkin's lymphoma (HL) and non-Hodgkin's lymphoma (NHL) occurring in Jordanian patients. A total of 55 patients with lymphoma were examined in this study. Of 55 patients, 30 and 25 were diagnosed as HL and NHL, respectively. The four HL subtypes were observed with the majority of the cases exhibited the mixed cellularity (MC) subtype followed by the nodular sclerosis (NS). The high grade was found to be the commonest subtype of NHL in our sample, followed by the low grade. The presence of EBV virus was detected by immunostating for expression of latent membrane protein-1 (LMP-1). The frequency of LMP-1 expression occurred more frequent in patients with HL (60.0%) than in patients with NHL (32.0%). The frequency of LMP-1 expression was also higher in patients with MC subtype (61.11%) than those patients with NS (28.57%). No age or gender difference in occurrence of EBV infection was observed among patient with HL. By contrast, the prevalence of EBV infection in NHL patients aged below 50 was lower (16.66%) than in NHL patients aged 50 or above (46.15%). In addition, EBV infection was more frequent in females with NHL (38.46%) than in male with NHL (25%). In NHL cases, the frequency of EBV infection in intermediate grade (60.0%) was high when compared with frequency of low (25%) or high grades (25%). In conclusion, analysis of LMP-1 expression indicates an important role for this viral oncogene in the pathogenesis of EBV-associated malignant lymphomas. These data also support the previous findings that people with EBV may develop lymphoma and that efforts to maintain low lymphoma should be considered for people with EBV infection.

Keywords: Hodgkin lymphoma, Epstein Barr virus, hematoxylin, infection, LMP-1 expression.

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21 Optimization of Growth Conditions for Acidic Protease Production from Rhizopus oligosporus through Solid State Fermentation of Sunflower Meal

Authors: Abdul Rauf, Muhammad Irfan, Muhammad Nadeem, Ishtiaq Ahmed, Hafiz Muhammad Nasir Iqbal

Abstract:

Rhizopus oligosporus was used in the present study for the production of protease enzyme under SSF. Sunflower meal was used as by-product of oil industry incorporated with organic salts was employed for the production of protease enzyme. The main purpose of the present was to study different parameters of protease productivity, its yields and to optimize basal fermentation conditions. The optimal conditions found for protease production using sunflower meal as a substrate in the present study were inoculum size (1%), substrate (Sunflower meal), substrate concentration (20 g), pH (3), cultivation period (72 h), incubation temperature (35oC), substrate to diluent-s ratio (1:2) and tween 81 (1 mL). The maximum production of protease in the presence of cheaper substrate at low concentration and stability at acidic pH, these characteristics make the strain and its enzymes useful in different industry.

Keywords: Acidic protease, Rhizopus oligosporus, Mediaoptimization, Solid state Fermentation

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20 Callusing in Stevia rebaudiana (Natural Sweetener) for Steviol Glycoside Production

Authors: Pratibha Gupta, Satyawati Sharma, Sanjay Saxena

Abstract:

Stevia rebaudiana Bertoni (natural sweetener) belongs to Asteraceae family and can be used as substitute of artificial sweeteners for diabetic patients. Conventionally, it is cultivated by seeds or stem cutting, but seed viability rate is poor. A protocol for callus induction and multiplication was developed to produce large no. of calli in short period. Surface sterilized nodal, leaf and root explants were cultured on Murashige and Skoog (MS) medium with different concentrations of plant hormone like, IBA, kinetin, NAA, 2,4-D, and NAA in combination with 2,4-D. 100% callusing was observed from leaf explants cultured on combination of NAA and 2,4-D after three weeks while with 2,4-D, only 10% callusing was observed. Calli obtained from leaf and root explants were shiny green while with nodal explants it was hard and brown. The present findings deal with induction of callusing in Stevia to achieve the rapid callus multiplication for study of steviol glycosides in callus culture.

Keywords: 2, 4-D, Callusing, NAA, Stevia, Steviol glycosides

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19 Estimating Enzyme Kinetic Parameters from Apparent KMs and Vmaxs

Authors: Simon Brown, Noorzaid Muhamad, David C Simcock

Abstract:

The kinetic properties of enzymes are often reported using the apparent KM and Vmax appropriate to the standard Michaelis-Menten enzyme. However, this model is inappropriate to enzymes that have more than one substrate or where the rate expression does not apply for other reasons. Consequently, it is desirable to have a means of estimating the appropriate kinetic parameters from the apparent values of KM and Vmax reported for each substrate. We provide a means of estimating the range within which the parameters should lie and apply the method to data for glutamate dehydrogenase from the nematode parasite of sheep Teladorsagia circumcincta.

Keywords: enzyme kinetics, glutamate dehydrogenase, intervalanalysis, parameter estimation.

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18 Effects of Chitosan as the Growth Stimulator for Grammatophyllum speciosum in Vitro Culture

Authors: Sopalun K., Thammasiri K., Ishikawa K.

Abstract:

The effects of chitosan, a biodegradable polymer, were studied in Grammatophyllum speciosum protocorm-like bodies (PLBs) in vitro culture. The chitosan concentration of 0, 5, 10, 15, 20, 25, 50 or 100 mg/l were supplemented in half-strength Murashige and Skoog (1/2 MS) liquid or on agar media containing 2% (w/v) sucrose. The results showed that liquid medium supplemented with 15 mg/l chitosan showed the highest relative growth rate (7-fold increase) of PLBs. On 1/2 MS agar medium supplemented with 25 mg/l chitosan gave the highest relative growth rate (4-fold increase). The relative growth rate of G. speciosum PLBs on agar medium was significantly lower than that in liquid medium. Moreover, chitosan, supplemented to agar medium promoted shoot formation but not rooting. However, supplementation at too high a level, such as 100 mg/l can inhibit growth and kill PLBs.

Keywords: Chitosan, Grammatophyllum speciosum, Growth stimulator

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17 An Information Theoretic Approach to Rescoring Peptides Produced by De Novo Peptide Sequencing

Authors: John R. Rose, James P. Cleveland, Alvin Fox

Abstract:

Tandem mass spectrometry (MS/MS) is the engine driving high-throughput protein identification. Protein mixtures possibly representing thousands of proteins from multiple species are treated with proteolytic enzymes, cutting the proteins into smaller peptides that are then analyzed generating MS/MS spectra. The task of determining the identity of the peptide from its spectrum is currently the weak point in the process. Current approaches to de novo sequencing are able to compute candidate peptides efficiently. The problem lies in the limitations of current scoring functions. In this paper we introduce the concept of proteome signature. By examining proteins and compiling proteome signatures (amino acid usage) it is possible to characterize likely combinations of amino acids and better distinguish between candidate peptides. Our results strongly support the hypothesis that a scoring function that considers amino acid usage patterns is better able to distinguish between candidate peptides. This in turn leads to higher accuracy in peptide prediction.

Keywords: Tandem mass spectrometry, proteomics, scoring, peptide, de novo, mutual information

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16 Biodegradation of Carbazole By a Promising Gram-Negative Bacterium

Authors: G. B. Singh, S. Srivastava, N. Gupta

Abstract:

In the present work we report a gram negative bacterial isolate, from soil of a dye industry, with promising biorefining and bioremediation potential. This isolate (GBS.5) could utilize carbazole (nitrogen containing polycyclic aromatic hydrocarbon) as the sole source of nitrogen and carbon and utilize almost 98% of 3mM carbazole in 100 hours. The specific activity of our GBS.5 isolate for carbazole degradation at 30°C and pH 7.0 was found to be 11.36 μmol/min/g dry cell weight as compared to 10.4 μmol/min/g dry cell weight, the highest reported specific activity till date. The presence of car genes (the genes involved in denitrogenation of carbazole) was confirmed through PCR amplification.

Keywords: Biodenitrogenation, Biorefining, Carbazoledegradation, Crude oil.

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15 Technique for Processing and Preservation of Human Amniotic Membrane for Ocular Surface Reconstruction

Authors: Irfan Z. Qureshi, Fareeha A., Wajid A. Khan

Abstract:

Human amniotic membrane (HAM) is a useful biological material for the reconstruction of damaged ocular surface. The processing and preservation of HAM is critical to prevent the patients undergoing amniotic membrane transplant (AMT) from cross infections. For HAM preparation human placenta is obtained after an elective cesarean delivery. Before collection, the donor is screened for seronegativity of HCV, Hbs Ag, HIV and Syphilis. After collection, placenta is washed in balanced salt solution (BSS) in sterile environment. Amniotic membrane is then separated from the placenta as well as chorion while keeping the preparation in BSS. Scrapping of HAM is then carried out manually until all the debris is removed and clear transparent membrane is acquired. Nitrocellulose membrane filters are then placed on the stromal side of HAM, cut around the edges with little membrane folded towards other side making it easy to separate during surgery. HAM is finally stored in solution of glycerine and Dulbecco-s Modified Eagle Medium (DMEM) in 1:1 ratio containing antibiotics. The capped borosil vials containing HAM are kept at -80°C until use. This vial is thawed to room temperature and opened under sterile operation theatre conditions at the time of surgery.

Keywords: HAM, AMT, ocular transplant

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14 SDS-induced Serine Protease Activity of an Antiviral Red Fluorescent Protein

Authors: Kalyankumar M. Matti, Chandrashekhar J. Savanurmath, Shivayogeppa B. Hinchigeri

Abstract:

A rare phenomenon of SDS-induced activation of a latent protease activity associated with the purified silkworm excretory red fluorescent protein (SE-RFP) was noticed. SE-RFP aliquots incubated with SDS for different time intervals indicated that the protein undergoes an obligatory breakdown into a number of subunits which exhibit autoproteolytic (acting upon themselves) and/or heteroproteolytic (acting on other proteins) activities. A strong serine protease activity of SE-RFP subunits on Bombyx mori nucleopolyhedrovirus (BmNPV) polyhedral protein was detected by zymography technique. A complete inhibition of BmNPV infection to silkworms was observed by the oral administration assay of the SE-RFP. Here, it is proposed that the SE-RFP prevents the initial infection of BmNPV to silkworms by obliterating the polyhedral protein. This is the first report on a silkworm red fluorescent protein that exhibits a protease activity on exposure to SDS. The present studies would help in understanding the antiviral mechanism of silkworm red fluorescent proteins.

Keywords: BmNPV, polyhedra, SE-RFP, SDS-induced protease activity, zymography.

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13 Effects of Intrauterine and Extrauterine Exposure to 1800 MHz GSM-Like Radiofrequency Radiation on Liver Regulatory Enzymes Activities in Infant Female Rabbits

Authors: A. Tomruk, G. Guler, B. Tandogan, E. Ozgur, N.N. Ulusu, N. Seyhan

Abstract:

In the present study, we aimed to design the intrauterine and extrauterine exposure to 1800 MHz GSM-like RF radiation and investigate its possible bio-effects on infant female rabbits. Totally thirty-six New Zealand White female rabbits, onemonth old, were randomly divided into four groups which are composed of 9 rabbits; i. Group I [Intrauterine (IU) exposure(-); Extrauterine (EU) exposure (-)], Group II [IU exposure (-); EU exposure (+)], Group III [IU exposure(+);EU exposure(-)], Group IV [IU exposure (+);EU exposure(+)]. The master regulatory enzymes activities of pentose phosphate pathway (glucose-6-phosphate dehydrogenase, G-6PD; 6-phosphogluconate dehydrogenase, 6- PGDH) and glutathione-dependent metabolism (glutathione peroxidase, GSH-Px; glutathione reductase, GR; glutathione Stransferase, GST, thioredoxin reductase, TRx) were analyzed in liver tissues of young female rabbits. Decreased G-6PD, 6-PGD, GSH-Px, GR activities were found in Group III compared to Group I (p<0.05, Mann Whitney). Increased GSH-px and TRx activities were found in Group IV compared to Group I (p<0.05, Mann Whitney). It can be concluded that the intrauterine and extrauterine exposure to GSMlike RF radiation may influence the liver regulatory enzymes activities.

Keywords: Radiofrequency (RF), intrauterine (IU) andextrauterine (EU) exposure, infant female rabbits.

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12 Effect of Dose Rate of Irradiation on Ultrastructure of Duodenal Mucosa

Authors: L. Labéjof, I. Mororó, P. Galle, G. Barbosa, M.I. Severo, A.H. de Oliveira

Abstract:

Ultrastructure of duodenum mucosa of irradiated rat was studied versus dose rate of irradiation following exposure to gamma rays from 60-Cobalt source. The animals were whole body irradiated at two dose rates (1 Gy.mn-1 and 1 Gy.h-1) and three total doses (1, 2 or 4 Gy) for each dose rate. 24 or 48 h after irradiation, their small intestine was removed and samples of duodenum were processed for observations under a transmission electron microscopy. Samples of duodenum mucosa of control rats were processed in the same way. For the lower dose rate of 1 Gy.h-1, main lesions characteristic of apoptosis were detected within irradiated enterocytes at a total dose of 2 Gy and 24 h after exposure. Necrosis was noted in the samples, 48 h after exposition. For the higher dose rate of 1 Gy.mn-1, fewer changes were detected at all total doses 24 or 48 h irradiation. Thus, it was shown that the appearance of radiationinduced alterations varies not only with increasing total dose and post-irradiation time but especially with decreasing dose rate.

Keywords: Dose rate, Radiation Inury, Apoptosis, SmallBowel

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11 A Novel Approach for Protein Classification Using Fourier Transform

Authors: A. F. Ali, D. M. Shawky

Abstract:

Discovering new biological knowledge from the highthroughput biological data is a major challenge to bioinformatics today. To address this challenge, we developed a new approach for protein classification. Proteins that are evolutionarily- and thereby functionally- related are said to belong to the same classification. Identifying protein classification is of fundamental importance to document the diversity of the known protein universe. It also provides a means to determine the functional roles of newly discovered protein sequences. Our goal is to predict the functional classification of novel protein sequences based on a set of features extracted from each protein sequence. The proposed technique used datasets extracted from the Structural Classification of Proteins (SCOP) database. A set of spectral domain features based on Fast Fourier Transform (FFT) is used. The proposed classifier uses multilayer back propagation (MLBP) neural network for protein classification. The maximum classification accuracy is about 91% when applying the classifier to the full four levels of the SCOP database. However, it reaches a maximum of 96% when limiting the classification to the family level. The classification results reveal that spectral domain contains information that can be used for classification with high accuracy. In addition, the results emphasize that sequence similarity measures are of great importance especially at the family level.

Keywords: Bioinformatics, Artificial Neural Networks, Protein Sequence Analysis, Feature Extraction.

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10 Fe3O4 and Fe3O4@Au Nanoparticles: Synthesis and Functionalisation for Biomolecular Attachment

Authors: Hendriëtte van der Walt, Lesley Chown, Richard Harris, Ndabenhle Sosibo, Robert Tshikhudo

Abstract:

The use of magnetic and magnetic/gold core/shell nanoparticles in biotechnology or medicine has shown good promise due to their hybrid nature which possesses superior magnetic and optical properties. Some of these potential applications include hyperthermia treatment, bio-separations, diagnostics, drug delivery and toxin removal. Synthesis refinement to control geometric and magnetic/optical properties, and finding functional surfactants for biomolecular attachment, are requirements to meet application specifics. Various high-temperature preparative methods were used for the synthesis of iron oxide and gold-coated iron oxide nanoparticles. Different surface functionalities, such as 11-aminoundecanoic and 11-mercaptoundecanoic acid, were introduced on the surface of the particles to facilitate further attachment of biomolecular functionality and drug-like molecules. Nanoparticle thermal stability, composition, state of aggregation, size and morphology were investigated and the results from techniques such as Fourier Transform-Infra Red spectroscopy (FT-IR), Ultraviolet visible spectroscopy (UV-vis), Transmission Electron Microscopy (TEM) and thermal analysis are discussed.

Keywords: Core/shell, Iron oxide, Gold coating, Nanoparticles.

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9 A Novel Multiplex Real-Time PCR Assay Using TaqMan MGB Probes for Rapid Detection of Trisomy 21

Authors: Mehrdad Hashemi, Mitra Behrooz Aghdam, Reza Mahdian, Ahmad Reza Kamyab

Abstract:

Cytogenetic analysis still remains the gold standard method for prenatal diagnosis of trisomy 21 (Down syndrome, DS). Nevertheless, the conventional cytogenetic analysis needs live cultured cells and is too time-consuming for clinical application. In contrast, molecular methods such as FISH, QF-PCR, MLPA and quantitative Real-time PCR are rapid assays with results available in 24h. In the present study, we have successfully used a novel MGB TaqMan probe-based real time PCR assay for rapid diagnosis of trisomy 21 status in Down syndrome samples. We have also compared the results of this molecular method with corresponding results obtained by the cytogenetic analysis. Blood samples obtained from DS patients (n=25) and normal controls (n=20) were tested by quantitative Real-time PCR in parallel to standard G-banding analysis. Genomic DNA was extracted from peripheral blood lymphocytes. A high precision TaqMan probe quantitative Real-time PCR assay was developed to determine the gene dosage of DSCAM (target gene on 21q22.2) relative to PMP22 (reference gene on 17p11.2). The DSCAM/PMP22 ratio was calculated according to the formula; ratio=2 -ΔΔCT. The quantitative Real-time PCR was able to distinguish between trisomy 21 samples and normal controls with the gene ratios of 1.49±0.13 and 1.03±0.04 respectively (p value <0.001). These results represent the presence of 3 copies of target gene in DS samples Vs 2 copies in normal controls. The results of quantitative Real-time PCR were in complete agreement with results of cytogenetic analysis. This study confirms previous reports regarding successful implementation of quantitative Real-time PCR for detection of trisomy 21. However, the assay has been improved by using MGB probes and more accurate data analysis. This assay, in particular, when performed in combination with another molecular assay such as QF-PCR or MLPA, can be used as a reliable technique for rapid prenatal diagnosis of trisomy 21.

Keywords: Trisomy 21, Real-time PCR, MGB-TaqMan Probes, Gene Dosage.

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8 Biosynthesis and In vitro Studies of Silver Bionanoparticles Synthesized from Aspergillusspecies and its Antimicrobial Activity against Multi Drug Resistant Clinical Isolates

Authors: M. Saravanan

Abstract:

Antimicrobial resistant is becoming a major factor in virtually all hospital acquired infection may soon untreatable is a serious public health problem. These concerns have led to major research effort to discover alternative strategies for the treatment of bacterial infection. Nanobiotehnology is an upcoming and fast developing field with potential application for human welfare. An important area of nanotechnology for development of reliable and environmental friendly process for synthesis of nanoscale particles through biological systems In the present studies are reported on the use of fungal strain Aspergillus species for the extracellular synthesis of bionanoparticles from 1 mM silver nitrate (AgNO3) solution. The report would be focused on the synthesis of metallic bionanoparticles of silver using a reduction of aqueous Ag+ ion with the culture supernatants of Microorganisms. The bio-reduction of the Ag+ ions in the solution would be monitored in the aqueous component and the spectrum of the solution would measure through UV-visible spectrophotometer The bionanoscale particles were further characterized by Atomic Force Microscopy (AFM), Fourier Transform Infrared Spectroscopy (FTIR) and Thin layer chromatography. The synthesized bionanoscale particle showed a maximum absorption at 385 nm in the visible region. Atomic Force Microscopy investigation of silver bionanoparticles identified that they ranged in the size of 250 nm - 680 nm; the work analyzed the antimicrobial efficacy of the silver bionanoparticles against various multi drug resistant clinical isolates. The present Study would be emphasizing on the applicability to synthesize the metallic nanostructures and to understand the biochemical and molecular mechanism of nanoparticles formation by the cell filtrate in order to achieve better control over size and polydispersity of the nanoparticles. This would help to develop nanomedicine against various multi drug resistant human pathogens.

Keywords: Bionanoparticles, UV-visible spectroscopy, AtomicForce Microscopy, Extracellular synthesis, Multi drug resistant, antimicrobial activity, Nanomedicine

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7 Characterization of an Acetobacter Strain Isolated from Iranian Peach that Tolerates High Temperatures and Ethanol Concentrations

Authors: K. Beheshti Maal, R. Shafiee

Abstract:

Vinegar is a precious food additive and complement as well as effective preservative against food spoilage. Recently traditional vinegar production has been improved using various natural substrates and fruits such as grape, palm, cherry, coconut, date, sugarcane, rice and balsam. These neoclassical fermentations resulted in several vinegar types with different tastes, fragrances and nutritional values because of applying various acetic acid bacteria as starters. Acetic acid bacteria include genera Acetobacter, Gluconacetobacter and Gluconobacter according to latest edition of Bergy-s Manual of Systematic Bacteriology that classifies genera on the basis of their 16s RNA differences. Acetobacter spp as the main vinegar starters belong to family Acetobacteraceae that are gram negative obligate aerobes, chemoorganotrophic bacilli that are oxidase negative and oxidize ethanol to acetic acid. In this research we isolated and identified a native Acetobacter strain with high acetic acid productivity and tolerance against high ethanol concentrations from Iranian peach as a summer delicious fruit that is very susceptible to food spoilage and decay. We used selective and specific laboratorial culture media such as Standard GYC, Frateur and Carr medium. Also we used a new industrial culture medium and a miniature fermentor with a new aeration system innovated by Pars Yeema Biotechnologists Co., Isfahan Science and Technology Town (ISTT), Isfahan, Iran. The isolated strain was successfully cultivated in modified Carr media with 2.5% and 5% ethanol simultaneously in high temperatures, 34 - 40º C after 96 hours of incubation period. We showed that the increase of ethanol concentration resulted in rising of strain sensitivity to high temperature. In conclusion we isolated and characterized a new Acetobacter strain from Iranian peach that could be considered as a potential strain for production of a new vinegar type, peach vinegar, with a delicious taste and advantageous nutritional value in food biotechnology and industrial microbiology.

Keywords: Acetobacter, Acetic Acid Bacteria, Vinegar, Peach, Food Biotechnology, Industrial Microbiology, Fermentation

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