Research Journal of Biotechnology

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Enhancement of Cognitive Activities of Codonopsislanceolata by Steaming Process associated with Ultra High Pressure Pretreatment

Choi Woon Yong and Lee Hyeon Yong

The cognitive-enhancing ability of Codonopsislanceolata was investigated by employing a consecutive steaming process followed by 300 MPa of ultra high pressure treatment. The neuroprotective effects on glutamate-induced cytotoxicity of PC12cells were estimated as 58.16% in adding 1 mg/mL of 70% ethanol extract. This extract also showed ca. 75% improvement of both cell growth and differentiation of PC12 cells by comparing with a positive control Donepezil as a nueroprotective agent. It was also found that the extract from this pretreatment process could effectively reduce both in vitro and in vivo gene expression of Acetylcholine Esterase that are closely related to cognitive effect. This conclusion was also supported by the results that the complex steaming process yielded higher amounts of total polyphenols and flavonoids as well less degradation of them. Further study would be necessary to reveal more detail mechanisms of its cognitive enhancing effect associated with antioxidant activities.

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Biochemical and haematological parameters and the impact of hypoalbuminaemia on mortality in carbapenem-resistant Acinetobacter baumannii complex bacteraemia

Chow S. F., Sheu D. C. and Liu C. P.

This study investigated the role of biochemical and haematological parameters for attributable mortality in patients with carbapenem-resistant Acinetobacter baumannii (CRAB) complex bacteraemia. From January 2009 to December 2012, patients with CRAB complex bacteraemia in a tertiary teaching hospital in Taiwan were enrolled retrospectively. We analyzed the characteristics and laboratory variables such as albumin, hemoglobin levels, white blood cell (WBC), platelet counts, creatinine and C-reactive protein (CRP) etc. 195 patients with CRAB complex bacteraemia were enrolled. After excluding incomplete medical records, laboratory parameters for 144 blood isolates were available for analysis. Independent risk factors for attributable mortality were intensive care unit (ICU) stay, the Acute Physiology and Chronic Health Evaluation II (APACHE II) score and respiratory tract source of bactaeremia and nonsurvivors had significantly low platelet counts, low albumin, low haemoglobin, elevated total bilirubin, elevated blood urea nitrogen (BUN) and elevated creatinine levels compared to survivors. The main finding of our study was that APACHE II score, respiratory tract source of bacteraemia and low albumin level were risk factors for attributable mortality in multivariate analysis. We suggest early treatment with albumin for hypoalbuminaemia in patients with CRAB complex bacteraemia.

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Identification of a novel endophytic Bacillus pumilus lipase from the seed of Pistacia chinensis Bunge

Song Chun-Zhu and Chen Dong-Hong

A lipase-producing endophytic bacterium PC1 was first isolated from seeds of woody energy plant Pistacia chinensis Bunge through tributyrin agar plate assay. Both morphological and molecular identification showed PC1 belongs to Bacillus pumilus strain, named Bacillus pumilus PC1. PC1 lipase displayed maximum activity at 350C and at pH 8, preferred to hydrolyze the substrates with medium-chain fatty acid and was found to be stable in low-temperature and alkaline conditions. The lipase activity was enhanced by K+ and Na+ and completely inhibited in the presence of Cu2+, Zn2+, Co2+ and surfactants. PC1 lipase displayed a relatively wide range of tolerance to organic solvents and methanol and acetone can notably improve its stability. Heterologous expression of PC1 lipase gene in E. coli produced a ~25kD band and GST-tag in its N-terminus had no effect on enzymatic secretion and activity. The novel PC1 lipase could be potentially applied to industrial and chemical fields.

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Glycerol transesterification with dimethyl carbonate using nano alumina based catalysts

Degirmenbasi Nebahat and Boz Nezahat

In this study, nanocrystalline alumina (γ-Al2O3) particles were functionalized by potassium carbonate (K2CO3) and used as alkaline heterogeneous catalysts for the transesterification of glycerol with dimethyl carbonate for the synthesis of glycerol carbonate. The loading ratio of K2CO3 to alumina (5-20 wt.%) and reaction temperature were investigated. Glycerol carbonate was successfully synthesized in the presence of synthesized catalyst with 20 wt.% K2CO3 loaded into alumina and the highest yield of glycerol carbonate of 94.5 % was obtained by 3 h of reaction time at 363 K (at reflux temperature of DMC) with a DMC/glycerol molar ratio of 2 and using 3 wt.% catalyst. Chemical analysis of the glycerol carbonate was performed by using ATR-FTIR and obtained glycerol carbonate was found to be pure.

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Assessment of Genetic Diversity among Indian Sesame (Sesamum indicum L.) Accessions using RAPD, ISSR and SSR Markers ‏

Kesawat Mahipal Singh,, Das Basanta Kumar, Dey Surjendu Kumar and Manorama

Three types of molecular markers RAPD, ISSR and SSR were used to study patterns of genetic variation among the 44 commercially cultivated sesame accessions representing different regions of the India. A high level of polymorphism was found with both RAPD and ISSR markers and the mean polymorphism information (PICs) content values were 0.130 and 0.675 for RAPD and ISSR markers respectively. In RAPD analyses, 120 out of 279 bands (43.01%) were polymorphic. The number of alleles ranged from 5 to 19 per primer with an average of 9.96 per primer. In ISSR analyses, a total of 183 alleles were detected among which 164 alleles (89.61%) were polymorphic. The number of alleles per primer ranged from 2 to 15 with an average of 7.32 alleles per ISSR primer. In addition, 12 selected SSR primers generated 41 amplified bands. The fragment size varied from 155 to 371 bp. Expected heterozygosity and PIC ranged from 0.430 to 0.780 and 0.404 to 0.740 respectively. A poor correlation (r = 0.107) was found between both sets of genetic similarity data, suggesting that both sets of markers revealed unrelated estimates of genetic relationships. Cluster analyses indicated that all 44 sesame accessions could be distinguished by RAPD, ISSR and SSR markers. The established RAPD, ISSR, and SSR markers would explore for further diversity analyses among a large commercially available of Indian sesame. Moreover, existence of genetic variability among the 44 commercially cultivated sesame accessions would be useful for mapping qualitative and quantitative trait, marker-assisted selection and a good starting point of sesame crop improvement programs in India.

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Comparative Study of Seed Protein Patterns of Oroxylum indicum (L.) Kurz. through SDS-PAGE

Talari Samatha, Neerati Raju, Yanamala Venkaiah and Nanna Rama Swamy

Electrophoretic study of two seed samples of Oroxylum indicum (L.) Kurz collected from two different regions of Warangal district (Telangana), India was undertaken using Sodium Dodecyl Sulphate Poly Acrylamide Gel Electrophoresis (SDS-PAGE). The results revealed the protein patterns resolved from aborted and fertile seeds of O. indicum indicate that there is a large variation of proteins present in aborted and fertile seeds collected from same region where as there is a similarity between the protein patterns of fertile seeds collected from both the regions. Thus, the present investigations indicate that there is no diversity based on SDS-PAGE proteins banding pattern in viable seeds of O. indicum collected from two different eco-climatic conditions of Warangal district whereas the difference in protein banding pattern was recorded between viable and non-viable seeds of O. indicum. Thus, based on our results, it can be concluded that for viability certain protein molecules are required which are absent in non-viable seeds. Probably the specific protein molecules related to DNA sequence may control the viability phenomenon in tree species.

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Arbuscular Mycorrhizal colonization in some plant species and relationship with the soil properties in the BCSIR campus of Chittagong, Bangladesh

Halder M., Dhar P.P., Nandi N.C. and Akhter S.

Study on the relationship between the soil properties and arbuscular mycorrhizal (AM) colonization of different plant species in the BCSIR Laboratories, Chittagong campus was carried out. Fine roots and rhizosphere soil of Acacia auriculiformis, Acalypha indica, Alpinia nigra, Artocarpus heterophyllus, Averrhoa carambola, Cinnamomum tamala, Elettaria cardamomum, Ficus benghalensis, Gardenia jasminoides, Hibiscus rosa-sinensis, Jasminum sambac, Manihot esculenta, Mimosa pudica, Mimusops elengi, Pinus palustris, Piper longum, Rauwolfia teraphylla, Sesbania sesban, Swietenia mahagoni and Tinospora cordifolia were collected, stained, processed and analyzed by established methods. Highest AM colonization was in G. jasminoides and the lowest was in S. sesban. No AM colonization occurred in P. palustris. There were no regular correlations among the soil properties and AM properties. Negative correlation between arbuscular mycorrhizal colonization soil phosphorus was significant (r=-0.646, p<0.01%). Significant and positive correlations between soil K contents, (r=0.826, p<0.01 for percent root colonization; r=0.601, p<0.01 for vesicular colonization and r=0.572, p<0.01 for arbuscular colonization) and AM properties were remarkable. EC was significantly related with root colonization (r=-0.535, p<0.05%). Vesicular colonization was significantly positive with the colonization. Independent influence of soil properties on the AM properties of host plant species for their nutritional acquisition in the field conditions was emphasized.

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Degradation of algae and cyanobacteria in water supply source using dielectric barrier discharge (DBD) plasma

Kim Sang-Don and Hyun Young-Jin

This work investigated a plasma water treatment based on a quartz tube and an alternating current (AC) and the application to the degradation of chlorophyll-a, green algae, diatom, phycocyanin, cyanobacreria (Microcystis aeruginosa and Anabaena affinis). The characteristics of dielectric barrier discharge (DBD) utilizing a quartz tube are to raise the gas-liquid interfacial area owing to the formation of numerous active species and instant transfer of short-lived species to water through the diffuser connected to the quartz tube right away after the generation. The current DBD process was found to effectively degrade not only algae but also cyanobacteria. It was suggested that the energy density needed to attain 60% of removal efficiency ranged from 380 to 9,700 J L-1 regarding the given compounds in the initial concentration range of 28 - 200 μg L-1. The experimental results exhibited that main factor being responsible for the efficient degradation was the types of the external and internal structure of these cyanobacteria and their initial concentration.

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Effect of Mutation S34N on hCAP in Periodontal Dental Arthritis Patients: Molecular Dynamics Simulation Study

Zhang Wei-Qun, Liu Yun-Qi, Ma Quan-Sheng, Wang Li and Hu Zhao-Hui

Studies have shown that genetic factors involved in the host responses might determine the severity of periodontitis and the missense mutation of S34N in Human cathelicidin antimicrobial peptide (hCAP) is one of the predominant factors in severity. The aim of this study is to study the effect of this mutation on protein dynamics. The FASTA sequence of the protein was subjected to four different (SIFT, Polyphen-2, PhD-SNP and MutPred) polymorphism effect prediction servers and the same format of the sequence was used to modeled the three dimensional structure of hCAP. The model structure was then subjected to Molecular dynamics simulations for Atomic insight. All the four servers predicted the mutation to be damaging and on analysing the effect of this mutation with the help of GROMACS, the results were showing the mutant structure unstable and differencing at every level in comparison with wild type hCAP.

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Genetic characterization of Aspergillus flavus and A. niger producing and non producing silver nanoparticles using DNA markers

Mahmoud Mohamed A., Abd-El-Aziz Abeer R. M. and Al-Othman Monira R.

Twelve Aspergilli isolates were including two species A. flavus and A. niger. A. flavus included six isolates, two isolates producing sliver nanoparticles (AgNPs) and four isolates non producing AgNPs. A. niger enclosed six isolates with a similar description. Random amplified polymorphic DNA (RAPD) and Inter simple sequence repeats (ISSR) DNA markers were used with the aim of genetically characterizing isolates of A. flavus and A. niger to discriminate between producing and non producing AgNPs isolates. RAPD and ISSR analysis revealed a high level of genetic diversity in the A. flavus and A. niger population useful for genetic characterization. A. flavus and A. niger isolates (producing and non producing AgNPs) are shown in RAPD and ISSR dendrogram with a random distribution. There was no clear-cut relationship between the RAPD and ISSR dendrogram and AgNPs production. RAPD and ISSR markers were not suitable to discriminate between producing and non producing AgNPs isolates.

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Screening and Characterization of a Salt-tolerant PAHs-degrading Strain isolated from Gasoline contaminated Soil

Zhang Xiangsheng

In this study, three PAHs (anthracene, phenanthrene and pyrene) were employed as substrates to isolate PAHs degrading microbial strains. A bacterial strain MN1 was screened out from soil sample nearby a bus gas station using plate screening techniques with PAHs as sole carbon and energy source. The bacterial strain could grow well on the screening agar plate with 800mg/L PAHs and 3.5% NaCl. With physiological and biochemical analyses and 16S rRNA gene sequence analysis, MN1 was identified as Pseudomonas stutzeri. MN1 performed well in degrading PAHs in aerobic shaking flasks. After 144h fermentation in shaking flasks, the biotic degradation rate of PAHs at 3.5% salinity by MN1 was 41.1% (anthracene and phenanthrene) and 44.6% (pyrene) respectively in 50mg/L-degrading culture media, with better degrading capacity of pyrene than that of anthracene and phenanthrene. This study gives hints for further study in PAHs aerobic biodegradation under saline conditions.

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Presence of segregation distortion in sheep

Al-Atiyat Raed M.

The main objective of this project was the investigation of presence of segregation distortion (SD) and description of other relevant parameters of multilocus genetics in Australian Merino sheep. The SD cases investigated three flocks of 98, 79 and 92 offspring and their DNA-based identified dams and sires. DNA samples were genotyped for 28 microsatellite (MS) markers located on different chromosomes. SD was estimated by studying the paternal segregation of alleles in offspring using a bootstrap procedure. The results showed a high proportion of studied loci which demonstrated significant SD. The results provided additional data which add to the common knowledge that sheep population structure and dynamics are affected by evolutionary forces such as gene flow, selection and many other random factors. These forces were noticed in the three populations. Finally, the results obtained from many loci in this project provided evidence of Mendelian violation. Moreover, they indicated that genes affecting male-related SD are spread over the genome.

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Effect of alcohol on release of green tea polyphenols from casein nanoparticles and its mathematical modeling

Ravi Theaj Prakash and Mandal Abul Kalam Azad

Since PalladoneTM was withdrawn from market after being reported the adverse drug reactions caused by alcohol induced dose dumping of hyromorphone, there has been concern on the risk of alcohol induced dose dumping. In this study, the potent influence of alcohol on green tea polyphenols (GTP) loaded casein nanoparticles was studied and the mechanism of drug release was determined. GTP was encapsulated into casein nanoparticles and release of GTP was carried out in phosphate buffer saline (PBS) alone or with 10, 20, 30 and 40% alcohol. As the concentration of alcohol in the medium increased, there was an increase in the cumulative percentage of drug release. Up to 97% of GTP was released in 48 h when the medium contained 40% alcohol while it was only 73% in PBS. The release data for all batches showed good correlation with zero order kinetics and mechanism of release was observed to be anomalous mode of drug transport. Concentration of alcohol showed no effect on the drug release mechanism and no dose dumping was observed. Thus, GTP loaded casein nanoparticles are safe from the danger of alcohol induced dose dumping and would serve as a potent carrier for the delivery of GTP.

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The latent infection pathways of Ophiocordyceps sinensis in Thitarodes larvae

Lei Wei, Shui Xiaorong, Zhang Guren and Liu Xin

Ophiocordyceps sinensis, an entomogenous fungus parasitic in the larvae of ghost moths (Lepidoptera), has always been used as one of the most valued Traditional Chinese Medicines throughout the Orient World. However, its occurrence and developmental mechanisms are crucial but almost completely unknown. Based on the real-time quantitative PCR method, the distribution of O. sinensis was determined in different tissues of the Thitarodes larvae including body-wall, fat-body, haemolymph and intestinal-wall. Meanwhile, two latent infection models based on the inward type starting at body-wall (R2 0.678) and the outward type starting at intestine (R2 0.271) were constructed successfully, the infection theory associated with food taking was supported after comparison and analyses. This study will contribute to deeply explain interaction between O. sinensis and its host Thitarodes insects.

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