Publications | Bioengineering and Life Sciences
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 394

World Academy of Science, Engineering and Technology

[Bioengineering and Life Sciences]

Online ISSN : 1307-6892

94 Loss of P16/INK4A Protein Expression is a Common Abnormality in Hodgkin's Lymphoma

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

Abstract:

P16/INK4A is tumor suppressor protein that plays a critical role in cell cycle regulation. Loss of P16 protein expression has been implicated in pathogenesis of many cancers, including lymphoma. Therefore, we sought to investigate if loss of P16 protein expression is associated with lymphoma and/or any specific lymphoma subtypes (Hodgkin-s lymphoma (HL) and nonHodgkin-s lymphoma (NHL)). Fifty-five lymphoma cases consisted of 30 cases of HL and 25 cases of NHL, with an age range of 3 to 78 years, were examined for loss of P16 by immunohistochemical technique using a specific antibody reacting against P16. In total, P16 loss was seen in 33% of all lymphoma cases. P16 loss was identified in 47.7% of HL cases. In contrast, only 16% of NHL showed loss of P16. Loss of P16 was seen in 67% of HL patients with 50 years of age or older, whereas P16 loss was found in only 42% of HL patients with less than 50 years of age. P16 loss in HL is somewhat higher in male (55%) than in female (30%). In subtypes of HL, P16 loss was found exclusively in all cases of lymphocyte depletion, lymphocyte predominance and unclassified cases, whereas P16 loss was seen in 39% of mixed cellularity and 29% of nodular sclerosis cases. In low grade NHL patients, P16 loss was seen in approximately one-third of cases, whereas no or very rare of P16 loss was found in intermediate and high grade cases. P16 loss did not show any correlation with age or gender of NHL patients. In conclusion, the high rate of P16 loss seen in our study suggests that loss of P16 expression plays a critical role in the pathogenesis of lymphoma, particularly with HL.

Keywords: B-cells, immunostaining, P16 protein, Reed-Sternberg cells, tumors.

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93 Isolation and Probiotic Characterization of Arsenic-Resistant Lactic Acid Bacteria for Uptaking Arsenic

Authors: Jatindra N. Bhakta, Kouhei Ohnishi, Yukihiro Munekage, Kozo Iwasaki

Abstract:

The growing health hazardous impact of arsenic (As) contamination in environment is the impetus of the present investigation. Application of lactic acid bacteria (LAB) for the removal of toxic and heavy metals from water has been reported. This study was performed in order to isolate and characterize the Asresistant LAB from mud and sludge samples for using as efficient As uptaking probiotic. Isolation of As-resistant LAB colonies was performed by spread plate technique using bromocresol purple impregnated-MRS (BP-MRS) agar media provided with As @ 50 μg/ml. Isolated LAB were employed for probiotic characterization process, acid and bile tolerance, lactic acid production, antibacterial activity and antibiotic tolerance assays. After As-resistant and removal characterizations, the LAB were identified using 16S rDNA sequencing. A total of 103 isolates were identified as As-resistant strains of LAB. The survival of 6 strains (As99-1, As100-2, As101-3, As102-4, As105-7, and As112-9) was found after passing through the sequential probiotic characterizations. Resistant pattern pronounced hollow zones at As concentration >2000 μg/ml in As99-1, As100-2, and As101-3 LAB strains, whereas it was found at ~1000 μg/ml in rest 3 strains. Among 6 strains, the As uptake efficiency of As102-4 (0.006 μg/h/mg wet weight of cell) was higher (17 – 209%) compared to remaining LAB. 16S rDNA sequencing data of 3 (As99- 1, As100-2, and As101-3) and 3 (As102-4, As105-7, and As112-9) LAB strains clearly showed 97 to 99% (340 bp) homology to Pediococcus dextrinicus and Pediococcus acidilactici, respectively. Though, there was no correlation between the metal resistant and removal efficiency of LAB examined but identified elevated As removing LAB would probably be a potential As uptaking probiotic agent. Since present experiment concerned with only As removal from pure water, As removal and removal mechanism in natural condition of intestinal milieu should be assessed in future studies.

Keywords: Lactic acid bacteria, As-resistant, characterization, Pediococcus sp., As removal probiotic.

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92 Effect of Electromagnetic Fields on Structure and Pollen Grains Development in Chenopodium album L

Authors: Leila Amjad, Mahsa Shafighi

Abstract:

The role of the pollen grain, with to the reproductive process of higher plants, is to deliver the spermatic cells to the embryo sac for egg fertilization. The aim of this project was study the effect of electromagnetic fields on structure and pollen grains development in Chenopodium album. Anthers of Chenopodium album L. were collected at different stages of development from control (without electromagnetic field) and plants grown at 10m from the field sources. Structure and development of pollen grains were studied and compared. The studying pollen structure by Light and Scanning electron microscopy showed that electromagnetic fields reduction of pollen grains number and male sterility, thus , in some anthers, pollen grains were attached together and deformed compared to control ones. The data presented suggest that prolonged exposures of plants to magnetic field may cause different biological effects at the cellular tissue and organ levels.

Keywords: Electromagnetic fields, pollen, Chenopodium albumL.

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91 Comparative Studies on Vertical Stratification,Floristic Composition, and Woody Species Diversity of Subtropical Evergreen Broadleaf Forests Between the Ryukyu Archipelago, Japan, and South China

Authors: M. Wu, S. M. Feroz, A. Hagihara, L. Xue, Z. L. Huang

Abstract:

In order to compare vertical stratification, floristic composition, and woody species diversity of subtropical evergreen broadleaf forests between the Ryukyu Archipelago, Japan, and South China, tree censuses in a 400 m2 plot in Ishigaki Island and a 1225 m2 plot in Dinghushan Nature Reserve were performed. Both of the subtropical forests consisted of five vertical strata. The floristic composition of the Ishigaki forest was quite different from that of the Dinghushan forest in terms of similarity on a species level (Kuno-s similarity index r0 = 0.05). The values of Shannon-s index H' and Pielou-s index J ' tended to increase from the bottom stratum upward in both forests, except H' for the top stratum in the Ishigaki forest and the upper two strata in the Dinghushan forest. The woody species diversity in the Dinghushan forest (H'= 3.01 bit) was much lower than that in the Ishigaki forest (H'= 4.36 bit).

Keywords: Floristic similarity, subtropical evergreen broadleaf forest, vertical stratification, woody species diversity.

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90 MIM: A Species Independent Approach for Classifying Coding and Non-Coding DNA Sequences in Bacterial and Archaeal Genomes

Authors: Achraf El Allali, John R. Rose

Abstract:

A number of competing methodologies have been developed to identify genes and classify DNA sequences into coding and non-coding sequences. This classification process is fundamental in gene finding and gene annotation tools and is one of the most challenging tasks in bioinformatics and computational biology. An information theory measure based on mutual information has shown good accuracy in classifying DNA sequences into coding and noncoding. In this paper we describe a species independent iterative approach that distinguishes coding from non-coding sequences using the mutual information measure (MIM). A set of sixty prokaryotes is used to extract universal training data. To facilitate comparisons with the published results of other researchers, a test set of 51 bacterial and archaeal genomes was used to evaluate MIM. These results demonstrate that MIM produces superior results while remaining species independent.

Keywords: Coding Non-coding Classification, Entropy, GeneRecognition, Mutual Information.

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89 A Comparison of SVM-based Criteria in Evolutionary Method for Gene Selection and Classification of Microarray Data

Authors: Rameswar Debnath, Haruhisa Takahashi

Abstract:

An evolutionary method whose selection and recombination operations are based on generalization error-bounds of support vector machine (SVM) can select a subset of potentially informative genes for SVM classifier very efficiently [7]. In this paper, we will use the derivative of error-bound (first-order criteria) to select and recombine gene features in the evolutionary process, and compare the performance of the derivative of error-bound with the error-bound itself (zero-order) in the evolutionary process. We also investigate several error-bounds and their derivatives to compare the performance, and find the best criteria for gene selection and classification. We use 7 cancer-related human gene expression datasets to evaluate the performance of the zero-order and first-order criteria of error-bounds. Though both criteria have the same strategy in theoretically, experimental results demonstrate the best criterion for microarray gene expression data.

Keywords: support vector machine, generalization error-bound, feature selection, evolutionary algorithm, microarray data

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88 Bioefficacy of Some Oil-Mixed Plant Derivatives against African Mud Catfish (Clarias gariepinus) Beetles, Dermestes maculatus and Necrobia rufipes

Authors: Akinwumi F. Olusegun

Abstract:

The efficacy of the separate mixing of four tropical spicy and medicinal plant products: Dennettia tripetala Baker (pepper fruit), Eugenia aromatica Hook (clove), Piper guineense (Schum and Thonn) (black pepper) and Monodora myristica (Dunal) (African nut-meg) with a household vegetable oil was evaluated under tropical storage conditions for the control and reproductive performance of Dermestes maculatus (De Geer) (hide beetle) and Necroba rufipes (De Geer) (copra beetle) on African catfish, Clarias gariepinus (Burchell). Each of the plant materials was pulverized into powder and applied as a mix of 1ml of oil and plant powder at 2.5, 5.0, 7.5 and 10.0g per 100g of dried fish, and allowed to dry for 6h. Each of the four oil-mixed powder treatments evoked significant (P < 05) mortalities of the two insects compared with the control (oil only) at 1, 3 and 7 days post treatment. The oil-powder mixture dosages did not prevent insect egg hatchability but while the emergent larvae on the treated samples died, the emergent larvae in the control survived into adults. The application of oil-mixed powders effectively suppressed the emergence of the larvae of the beetles. Similarly, each of the oil-powder mixtures significantly reduced weight loss in smoked fish that were exposed to D. maculatus and N. rufipes when compared to the control (P < 05). The results of this study suggest that the plant powders rather than the domestic oil demonstrated protective ability against the fish beetles and confirm the efficacy of the plant products as pest control agents.

Keywords: Catfish, Fish beetles, Fish preservation, Oil-powder mix, Plant products.

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87 X-ray Crystallographic Analysis of MinC N-Terminal Domain from Escherichia coli

Authors: Jun Yop An, Kyoung Ryoung Park, Jung-Gyu Lee, Hyung-Seop Youn, Jung-Yeon Kang, Gil Bu Kang, Soo Hyun Eom

Abstract:

MinC plays an important role in bacterial cell division system by inhibiting FtsZ assembly. However, the molecular mechanism of the action is poorly understood. E. coli MinC Nterminus domain was purified and crystallized using 1.4 M sodium citrate pH 6.5 as a precipitant. X-ray diffraction data was collected and processed to 2.3 Å from a native crystal. The crystal belonged to space group P212121, with the unit cell parameters a = 52.7, b = 54.0, c = 64.7 Å. Assuming the presence of two molecules in the asymmetric unit, the Matthews coefficient value is 1.94 Å3 Da-1, which corresponds to a solvent content of 36.5%. The overall structure of MinCN is observed as a dimer form through anti-parallel ß-strand interaction.

Keywords: MinC, Cell division, Crystallization.

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86 Chitosan/Casein Microparticles: Preparation, Characterization and Drug Release Studies

Authors: Selvakumar Dhanasingh, Shunmuga Kumar Nallaperumal

Abstract:

Microparticles carrier systems made from naturally occurring polymers based on chitosan/casein system appears to be a promising carrier for the sustained release of orally and parenteral administered drugs. In the current study we followed a microencapsulation technique based aqueous coacervation method to prepare chitosan/casein microparticles of compositions 1:1, 1:2 and 1:5 incorporated with chloramphenicol. Glutaraldehyde was used as a chemical cross-linking agent. The microparticles were prepared by aerosol method and studied by optical microscopy, infrared spectroscopy, thermo gravimetric analysis, swelling studies and drug release studies at various pH. The percentage swelling of the polymers are found to be in the order pH 4 > pH 10 > pH 7 and the increase in casein composition decrease the swelling percentage. The drug release studies also follow the above order.

Keywords: Chitosan/casein micro particles, chloramphenicol, drug release, microencapsulation.

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

Authors: J.Yojina, W. Ngamsaad, N. Nuttavut, D.Triampo, Y. Lenbury, W. Triampo, P. Kanthang, S.Sriyab

Abstract:

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.

Keywords: lattice Boltzmann method, cell division, Minproteins oscillation, nucleoid

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

Authors: Uun Yanuhar

Abstract:

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

Keywords: C.altivelis, immunogenic, MHC, V.alginolyticus.

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83 Cell Growth and Metabolites Produced by Fluorescent Pseudomonad R62 in Modified Chemically Defined Medium

Authors: K. Saharan, M.V. R. K. Sarma, A. S. Roesti, A. Prakash, B. N. Johri, M. Aragno, V. S. Bisaria, V. Sahai

Abstract:

Chemically defined Schlegel-s medium was modified to improve production of cell growth and other metabolites that are produced by fluorescent pseudomonad R62 strain. The modified medium does not require pH control as pH changes are kept within ± 0.2 units of the initial pH 7.1 during fermentation. The siderophore production was optimized for the fluorescent pseudomonad strain in the modified medium containing 1% glycerol as a major carbon source supplemented with 0.05% succinic acid and 0.5% Ltryptophan. Indole-3 acetic acid (IAA) production was higher when L-tryptophan was used at 0.5%. The 2,4- diacetylphloroglucinol (DAPG) was higher with amended three trace elements in medium. The optimized medium produced 2.28 g/l of dry cell mass and 900 mg/l of siderophore at the end of 36 h cultivation, while the production levels of IAA and DAPG were 65 mg/l and 81 mg/l respectively at the end of 48 h cultivation.

Keywords: Fluorescent pseudomonad, Fermentation, Metabolites production, PGPR.

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82 Biodegradation of Cyanide by a Novel Cyanidedegrading Bacterium

Authors: S. Potivichayanon, R. Kitleartpornpairoat

Abstract:

The objectives were to identify cyanide-degrading bacteria and study cyanide removal efficiency. Agrobacterium tumefaciens SUTS 1 was isolated. This is a new strain of microorganisms for cyanide degradation. The maximum growth rate of SUTS 1 obtained 4.7 × 108 CFU/ml within 4 days. The cyanide removal efficiency was studied at 25, 50, and 150 mg/L cyanide. The residual cyanide, ammonia, nitrate, nitrite, pH, and cell counts were analyzed. At 25 and 50 mg/L cyanide, SUTS 1 obtained similar removal efficiency approximately 87.50%. At 150 mg/L cyanide, SUTS 1 enhanced the cyanide removal efficiency up to 97.90%. Cell counts of SUTS 1 increased when the cyanide concentration was set at lower. The ammonia increased when the removal efficiency increased. The nitrate increased when the ammonia decreased but the nitrite did not detect in all experiments. pH values also increased when the cyanide concentrations were set at higher.

Keywords: Biodegradation, Cyanide-degrading bacteria, Removal efficiency, Residual cyanide

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81 Toxicity of Copper and Cadmium to Freshwater Fishes

Authors: M. Shuhaimi-Othman, Y. Nadzifah, A.K. Ahmad

Abstract:

Two freshwater fishes, Rasbora sumatrana (Cyprinidae) and Poecilia reticulata (guppy) (Poeciliidae) were exposed for a four-day period in the laboratory condition to a range of copper (Cu) and cadmium (Cd) concentrations. Mortality was assessed and median lethal concentrations (LC50) were calculated. LC50 increased with decrease in mean exposure times for both metals. For R. sumatrana, LC50s for 24, 48, 72 and 96 hours for Cu were 54.2, 30.3, 18.9 and 5.6 μg/L and for Cd 1440.2, 459.3, 392.3 and 101.6 μg/L respectively. For P. reticulata, LC50s for 24, 48, 72 and 96 hours for Cu were 348.9, 145.4, 61.3 and 37.9 μg/L and for Cd 8205.6, 2827.1, 405.8 and 168.1 μg/L, respectively. Results indicated that the Cu was more toxic than Cd to both fishes (Cu>Cd) and R. sumatrana was more sensitive than P. reticulata to the metals.

Keywords: Acute, heavy metals, LT50, toxicity

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80 The Potential of Strain M Protease in Degradations of Protein in Natural Rubber Latex

Authors: Norlin Pauzi, Ahmad R.M. Yahya, Zairossani Nor, Amirul A. Abdullah

Abstract:

Strain M was isolated from the latex of Hevea brasiliensis that grow in the rubber farm area of Malaysia Rubber Board. Strain M was tentatively identified as Bacillus sp. Strain M demonstrated high protease production at pH 9, and this was suitable to be applied in rubber processing that was in alkaline conditions. The right and suitable proportion to be used in applying supernatant into the latex was two parts of latex and one part of enzyme. In this proportion, the latex was stable throughout the 72 hours of treatment. The potential of strain M to degrade protein in the natural rubber latex was proven with the reduction of 79.3% nitrogen in 24 hours treatment. Centrifugation process of the latex before undergoing the treatment had increased the protein degradation in latex. Although the centrifugation process did not achieve zero nitrogen content, it had improved the performance of protein denaturing in the natural rubber.

Keywords: Hevea brasiliensis, Bacillus sp., protease, latex.

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79 Inhibitory Effect of Lactic Acid and Nisin on Bacterial Spoilage of Chilled Shrimp

Authors: A. R. Shirazinejad, I. Noryati, A. Rosma, I. Darah

Abstract:

Lactic acid alone and its combined application with nisin were evaluated for reducing population of naturally occurring microorganisms on chilled shrimp. Fresh shrimps were dipped in 0, 1.0% and 2.0% (v/v) lactic acid alone and their combined application with 0.04 (g/L/kg) nisin solution for 10 min. Total plate counts of aerobic bacteria (TPCs), Psychrotrophic counts, population of Pseudomonas spp., H2S producing bacteria and Lactic acid bacteria (LAB) on shrimps were determined during storage at 4 °C. The results indicated that total plate counts were 2.91 and 2.63 log CFU/g higher on untreated shrimps after 7 and 14 days of storage, respectively, than on shrimps treated with 2.0% lactic acid combined with 0.04 (g/L/kg) nisin. Both concentrations of lactic acid indicated significant reduction on Pseudomonas counts during storage, while 2.0% lactic acid combined with nisin indicated the highest reduction. In addition, H2S producing bacteria were more sensitive to high concentration of lactic acid combined with nisin during storage.

Keywords: Shrimp, lactic acid, nisin, spoilage bacteria

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78 Influence of Static Pressure on Viability of Entomopathogenic Nematodes – Steinernema feltiae

Authors: J. Chojnacki, E. Dulcet, A. Grieger

Abstract:

The entomopathogenic nematodes Steinernema feltiaeare are components of many biological pesticides. The biological pesticides are applicated by means a spraying machines. The influence of high pressure operating time on viability of nematodes has been experimentally investigated in order to explain if static pressure inside of the sprayers installation was able to destroy nematodes. The value of pressure was 55 MPa and its maximum operating time was 3 hours. Changes were found in viability of pressurized samples of nematodes, mixed with water.

Keywords: Entomopathogenic nematodes, biopesticides, highpressure, sprayer.

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77 Identification Characterization and Production of Phytase from Endophytic Fungi

Authors: Yetti Marlida , Rina Delfita , Neni Gusmanizar, Gita Ciptaan

Abstract:

Phytases are acid phosphatase enzymes, which efficiently cleave phosphate moieties from phytic acid, thereby generating myo-inositol and inorganic phosphate. Thirty four isolates of endophytic fungi to produce of phytases were isolated from leaf, stem and root fragments of soybean. Screening of 34 isolates of endophytic fungi identified the phytases produced by Rhizoctonia sp. and Fusarium verticillioides . The phytase production were the best induced by phytic acid and rice bran compared the others inducer in submerged fermentation medium used. The phytase produced by both Rhizoctonia sp. and F. verticillioides have pH optimum at 4.0 and 5.0 respectively. The characterization of phytase from Fusarium verticillioides showed that temperature optimum was 500C and stability until 600C, the pH optimum 5.0 and pH stability was 2.5 – 6.0, and substrate specificity were rice bran>soybean meal>corn> coconut cake, respectively.

Keywords: endophytic fungus, phytase, soybean, Rhizoctoniasp., Fusarium verticillioides,

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76 Direct and Indirect Somatic Embryogenesis from Petiole and Leaf Explants of Purple Fan Flower (Scaevola aemula R. Br. cv. 'Purple Fanfare')

Authors: Shyama Ranjani Weerakoon

Abstract:

Direct and indirect somatic embryogenesis (SE) from petiole and leaf explants of Scaevola aemula R. Br. cv. 'Purple Fanfare' was achieved. High frequency of somatic embryos was obtained directly from petiole and leaf explants using an inductive plant growth regulator signal thidiazuron (TDZ). Petiole explants were more responsive to SE than leaves. Plants derived from somatic embryos of petiole explants germinated more readily into plants. SE occurred more efficiently in half-strength Murashige and Skoog (MS) medium than in full-strength MS medium. Non-embryogenic callus induced by 2, 4-dichlorophenoxyacetic acid was used to investigate the feasibility of obtaining SE with TDZ as a secondary inductive plant growth regulator (PGR) signal. Non-embryogenic callus of S. aemula was able to convert into an “embryogenic competent mode" with PGR signal. Protocol developed for induction of direct and indirect somatic embryogenesis in S. aemula can improve the large scale propagation system of the plant in future.

Keywords: Petiole and leaf explants, Scaevola aemula, Somaticembryogenesis

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75 Structural Basis of Resistance of Helicobacterpylori DnaK to Antimicrobial Peptide Pyrrhocoricin

Authors: Musammat F. Nahar, Anna Roujeinikova

Abstract:

Bacterial molecular chaperone DnaK plays an essential role in protein folding, stress response and transmembrane targeting of proteins. DnaKs from many bacterial species, including Escherichia coli, Salmonella typhimurium and Haemophilus infleunzae are the molecular targets for the insect-derived antimicrobial peptide pyrrhocoricin. Pyrrhocoricin-like peptides bind in the substrate recognition tunnel. Despite the high degree of crossspecies sequence conservation in the substrate-binding tunnel, some bacteria are not sensitive to pyrrhocoricin. This work addresses the molecular mechanism of resistance of Helicobacter pylori DnaK to pyrrhocoricin. Homology modelling, structural and sequence analysis identify a single aminoacid substitution at the interface between the lid and the β-sandwich subdomains of the DnaK substrate-binding domain as the major determinant for its resistance.

Keywords: Helicobacter pylori, molecular chaperone DnaK, pyrrhocoricin, structural biology.

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74 Communities of Ammonia-oxidizing Archaea and Bacteria in Enriched Nitrifying Activated Sludge

Authors: Puntipar Sonthiphand, Tawan Limpiyakorn

Abstract:

In this study, communities of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) in nitrifying activated sludge (NAS) prepared by enriching sludge from a municipal wastewater treatment plant in three continuous-flow reactors receiving an inorganic medium containing different ammonium concentrations of 2, 10, and 30 mM NH4 +-N (NAS2, NAS10, and NAS30, respectively) were investigated using molecular analysis. Results suggested that almost all AOA clones from NAS2, NAS10, and NAS30 fell into the same AOA cluster and AOA communities in NAS2 and NAS10 were more diverse than those of NAS30. In contrast to AOA, AOB communities obviously shifted from the seed sludge to enriched NASs and in each enriched NAS, communities of AOB varied particularly. The seed sludge contained members of N. communis cluster and N. oligotropha cluster. After it was enriched under various ammonium loads, members of N. communis cluster disappeared from all enriched NASs. AOB with high affinity to ammonia presented in NAS 2, AOB with low affinity to ammonia presented in NAS 30, and both types of AOB survived in NAS 10. These demonstrated that ammonium load significantly influenced AOB communities, but not AOA communities in enriched NASs.

Keywords: ammonia-oxidizing bacteria, ammonia-oxidizingarchaea, nitrifying activated sludge.

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73 Function of miR-125b in Zebrafish Neurogenesis

Authors: Minh T. N. Le, Cathleen Teh, Ng Shyh-Chang, Vladimir Korzh, Harvey F. Lodish, Bing Lim

Abstract:

MicroRNAs are an important class of gene expression regulators that are involved in many biological processes including embryogenesis. miR-125b is a conserved microRNA that is enriched in the nervous system. We have previously reported the function of miR-125b in neuronal differentiation of human cell lines. We also discovered the function of miR-125b in regulating p53 in human and zebrafish. Here we further characterize the brain defects in zebrafish embryos injected with morpholinos against miR-125b. Our data confirm the essential role of miR-125b in brain morphogenesis particularly in maintaining the balance between proliferation, cell death and differentiation. We identified lunatic fringe (lfng) as an additional target of miR-125b in human and zebrafish and suggest that lfng may mediate the function of miR-125b in neurogenesis. Together, this report reveals new insights into the function of miR- 125b during neural development of zebrafish.

Keywords: microRNA, miR-125b, neurogenesis, zebrafish.

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72 Biological Characterization of the New Invasive Brine Shrimp Artemia franciscana in Tunisia: Sabkhet Halk El-Menzel

Authors: Hachem Ben Naceur, Amel Ben Rejeb Jenhani, Mohamed Salah Romdhane

Abstract:

Endemic Artemia franciscana populations can be found throughout the American continent and also as an introduced specie in several country all over the world, such as in the Mediterranean region where Artemia franciscana was identified as an invasive specie replacing native Artemia parthenogenetica and Artemia salina. In the present study, the characterization of the new invasive Artemia franciscana reported from Sabkhet Halk El-Menzel (Tunisia) was done based on the cysts biometry, nauplii instar-I length, Adult sexual dimorphism and fatty acid profile. The mean value of the diameter of non-decapsulated and decapsulated cysts, chorion thickness and naupliar length is 235.8, 226.3, 4.75 and 426.8 μm, respectively. Sexual dimorphism for adults specimen showed that maximal distance between compound eyes, diameter for compound eyes, length of first antenna and the abdomen length compared to the total body length ratio, are the most important variables for males and females discrimination with a total contribution of 62.39 %. The analysis of fatty acid methyl esters profile of decapsulated cysts resulted in low levels of linolenic acid (LLA, C18:3n-3) and high levels of eicosapentaenoic acid (EPA, C20:5n-3) with 3.11 and 11.10 %, respectively. Low quantity of docosahexaenoic acid (DHA, 22:6n-3) was also observed with 0.17 mg.g-1 dry weight.

Keywords: Invasive Artemia franciscana, biometry, sexualdimorphism, fatty acid, Tunisia.

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71 Development of All-male Fingerlings by Heat Treatment and the Genetic Mechanism of Heat Induced Sex Determination in Nile Tilapia(Oreochromis niloticus L.)

Authors: P. O. Angienda, B. O. Aketch, E. N. Waindi

Abstract:

Juvenile Nile tilapia subjected to heat treatment at temperatures ranging from 260C to 370C showed positive correlation (P<0.01) between treatment temperatures and resultant sex ratios, while, survival rate of the fry showed a negative correlation against temperature (P<0.01). The optimal temperature for both sex shift towards males and survival rates was 36±0.5°C, producing male percentage of 86.31 and a fry survival of 65.25. To determine the genetic basis of temperature sex-determination in Nile tilapia, we employed three microsatellite markers (Abur36, Abur100 and UNH846). Abur36 predicted the sex of 95% of the heat induced individuals, suggesting that the locus influence sex ratio and its interaction with temperature result in male biased sex ratio. This locus could turn out to be the major sex determining gene operating in Nile tilapia. These markers could be used in marker-assisted selection to select genotypes that give a higher percentage of males for commercial production.

Keywords: Heat treatment, Microsatellite, Nile tilapia, sex-determination.

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70 Human Elastin-derived Biomimetic Coating Surface to Support Cell Growth

Authors: Antonella Bandiera

Abstract:

A new sythetic gene coding for a Human Elastin-Like Polypeptide was constructed and expressed. The recombinant product was tested as coating agent to realize a surface suitable for cell growth. Coatings showed peculiar features and different human cell lines were seeded and cultured. All cell lines tested showed to adhere and proliferate on this substrate that has been shown also to exert a specific effect on cells, depending on cell type.

Keywords: elastin, recombinant protein, coating, cell adhesion.

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

Authors: Sarkar M. A. Kawsar, Sarkar M. A. Mamun, Md S. Rahman, Hidetaro Yasumitsu, Yasuhiro Ozeki

Abstract:

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

Keywords: Antibacterial, Halichondria okadai, Inhibition zone, Lectin.

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68 Apoptosis Induced by Low-concentration Ethanol in Hepatocellular Carcinoma Cell Strains and Down-regulated AFP and Survivin Analysis by Proteomic Technology

Authors: Xin Kai, Juan Li, Sexin Huang, Zengliang Bai

Abstract:

Ethanol is generally used as a therapeutic reagent against Hepatocellular carcinoma (HCC or hepatoma) worldwide, as it can induce Hepatocellular carcinoma cell apoptosis at low concentration through a multifactorial process regulated by several unknown proteins. This paper provides a simple and available proteomic strategy for exploring differentially expressed proteins in the apoptotic pathway. The appropriate concentrations of ethanol required to induce HepG2 cell apoptosis were first assessed by MTT assay, Gisma and fluorescence staining. Next, the central proteins involved in the apoptosis pathway processs were determined using 2D-PAGE, SDS-PAGE, and bio-software analysis. Finally the downregulation of two proteins, AFP and survivin, were determined by immunocytochemistry and reverse transcriptase PCR (RT-PCR) technology. The simple, useful method demonstrated here provides a new approach to proteomic analysis in key bio-regulating process including proliferation, differentiation, apoptosis, immunity and metastasis.

Keywords: Hepatocellular carcinoma, Ethanol, Proteomics, survivin and AFP

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67 Inhibition of the Growth of Pathogenic Candida spp. by Salicylhydroxamic Acid

Authors: Shu-Ying Marissa Pang, Stephen Tristram, Simon Brown

Abstract:

Candida spp. are common and aggressive pathogens. Because of the growing resistance of Candida spp. to current antifungals, novel targets, found in Candida spp. but not in humans or other flora, have to be identified. The alternative oxidase (AOX) is one such possibility. This enzyme is insensitive to cyanide, but is sensitive to compounds such as salicylhydroxamic acid (SHAM), disulfiram and n-alkyl gallates. The growth each of six Candida spp. was inhibited significantly by ~13 mM SHAM or 2 mM cyanide, albeit to differing extents. In C. dubliniensis, C. krusei and C. tropicalis the rate of O2 uptake was inhibited by 18-36% by 25 mM SHAM, but this had little or no effect on C. glabrata, C. guilliermondii or C. parapsilosis. Although SHAM substantially inhibited the growth of Candida spp., it is unlikely that the inhibition of AOX was the cause. Salicylhydroxamic acid is used therapeutically in the treatment of urinary tract infections and urolithiasis, but it also has some potential in the treatment of Candida spp. infection.

Keywords: alternative oxidase, Candida spp., growth, respiration, salicylhydroxamic acid.

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

Authors: Le Thuy Mai, Vu Nguyen Thanh

Abstract:

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

Keywords: β-Glucosidase, Pichia pastoris, Saccharomycopsisfibuligera, recombinant enzyme.

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65 A New Method for Rapid DNA Extraction from Artemia (Branchiopoda, Crustacea)

Authors: R. Manaffar, R. Maleki, S. Zare, N. Agh, S. Soltanian, B. Sehatnia, P. Sorgeloos, P. Bossier, G. Van Stappen

Abstract:

Artemia is one of the most conspicuous invertebrates associated with aquaculture. It can be considered as a model organism, offering numerous advantages for comprehensive and multidisciplinary studies using morphologic or molecular methods. Since DNA extraction is an important step of any molecular experiment, a new and a rapid method of DNA extraction from adult Artemia was described in this study. Besides, the efficiency of this technique was compared with two widely used alternative techniques, namely Chelex® 100 resin and SDS-chloroform methods. Data analysis revealed that the new method is the easiest and the most cost effective method among the other methods which allows a quick and efficient extraction of DNA from the adult animal.

Keywords: APD, Artemia, DNA extraction, Molecularexperiments

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