Search results for: Extracellular%20laccase

Authors: P. Chaijak, M. Lertworapreecha, C. Sukkasem

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Extracellular laccases are copper-containing microbial enzymes with many industrial biotechnological applications. This study evaluated the ability of nutrients in coconut coir to enhance the yield of extracellular laccase of Galactomyces reesii IFO 10823 and develop a co-culture between this yeast and other filamentous fungi isolated from the fungus comb of Macrotermes sp. The co-culture between G. reesii IFO 10823 and M. indicus FJ-M-5 (G3) gave the highest activity at 580.20 U/mL. When grown in fermentation media prepared from coconut coir and distilled water at 70% of initial moisture without supplement addition, G3 produced extracellular laccase of 113.99 U/mL.

Keywords: Extracellular laccase, production, yeast, natural inducer.

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Authors: Hanina M. N., Hairul Shahril M., Ismatul Nurul Asyikin I., Abdul Jalil A. K., Salina M. R., Maryam M. R., Rosfarizan M.

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The extracellular proteins secreted by bacteria may be increased in stressful surroundings, such as in the presence of antibiotics. It appears that many antibiotics, when used at low concentrations, have in common the ability to activate or repress gene transcription, which is distinct from their inhibitory effect. There have been comparatively few studies on the potential of antibiotics as a specific chemical signal that can trigger a variety of biological functions. Therefore, this study was carried out to determine the effect of Streptomycin Sulfate in regulating extracellular proteins secreted by Bacillus subtilis ATCC21332. Results of Microdilution assay showed that the Minimum Inhibition Concentration (MIC) of Streptomycin Sulfate on B. subtilis ATCC21332 was 2.5 mg/ml. The bacteria cells were then exposed to Streptomycin Sulfate at concentration of 0.01 MIC before being further incubated for 48h to 72 h. The extracellular proteins secreted were then isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Proteins profile revealed that three additional bands with approximate sizes of 30 kDa, 22 kDa and 23 kDa were appeared for the treated bacteria with Streptomycin Sulfate. Thus, B. subtilis ATCC21332 in stressful condition with the presence of Streptomycin Sulfate at low concentration could induce the extracellular proteins secretion.

Keywords: Bacillus subtilis ATCC21332, Streptomycin Sulfate, extracellular proteins.

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Authors: Rusudan Sujashvili, Ekaterine Bakuradze, Irina Modebadze, Davit Dekanoidze

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Extracellular ubiquitin in vivo effect on regenerative liver cells and liver histoarchitectonics has been studied. Experiments were performed on mature female white rats. Partial hepatectomy was made using the modified method of Higgins and Anderson. Standard histopathological assessment of liver tissue was used. Proliferative activity of hepatocytes was analyzed by colchicine mitotic index and immunohistochemical staining on ki67. We have found that regardless of number of injections and dose of extracellular ubiquitin liver histology has not been changed, so at tissue level no effect was observed. In vivo double injection of ubiquitin significantly decreases the mitotic activity at 32 hour point after partial hepatectomy. Thus, we can conclude that in vivo injected extracellular ubiquitin inhibits proliferative activity of hepatocytes in partially hepatectomyzed rats.

Keywords: Liver, regeneration, proliferation, ubiquitin.

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Authors: Yue Du, Sahan C. B. Herath, Qing-Guo Wang, Harry Asada, Peter C. Y. Chen

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Mechanical interaction between endothelial cells (ECs) and the extracellular matrix (or collagen gel) is known to influence the sprouting response of endothelial cells during angiogenesis. This influence is believed to impact on the capability of endothelial cells to sense soluble chemical cues. Quantitative analysis of endothelial-cell-mediated displacement of the collagen gel provides a means to explore this mechanical interaction. Existing analysis in this context is generally limited to 2D settings. In this paper, we investigate the mechanical interaction between endothelial cells and the extracellular matrix in terms of the endothelial-cellmediated displacement of the collagen gel in both 2D and 3D. Digital image correlation and Digital volume correlation are applied on confocal reflectance image stacks to analyze cell-mediated displacement of the gel. The skeleton of the sprout is extracted from phase contrast images and superimposed on the displacement field to further investigate the link between the development of the sprout and the displacement of the gel.

Keywords: Angiogenesis, digital image correlation, digital volume correlation, interaction between ECs and ECM.

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Authors: Gopalu Karunakaran, Matheswaran Jagathambal, Nguyen Van Minh, Evgeny Kolesnikov, Denis Kuznetsov

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This work investigated the use of Beijerinckia indica extracellular extract for the synthesis of silver nanoparticles using AgNO₃. The formation of nanoparticles was confirmed by different methods, such as UV-Vis absorption spectroscopy, XRD, FTIR, EDX, and TEM analysis. The formation of silver nanoparticles (AgNPs) was confirmed by the change in color from light yellow to dark brown. The absorbance peak obtained at 430 nm confirmed the presence of silver nanoparticles. The XRD analysis showed the cubic crystalline phase of the synthesized nanoparticles. FTIR revealed the presence of groups that acts as stabilizing and reducing agents for silver nanoparticles formation. The synthesized silver nanoparticles were generally found to be spherical in shape with size ranging from 5 to 20 nm, as evident by TEM analysis. These nanoparticles were found to inhibit pathogenic bacterial strains. This work proved that the bacterial extract is a potential eco-friendly candidate for the synthesis of silver nanoparticles with promising antibacterial and antioxidant properties. 

Keywords: Antioxidant activity, antimicrobial activity, Beijerinckia indica, characterisation, extracellular extracts, silver nanoparticles.

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Authors: Richard Grünes, Karel Roubik

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The article deals with technical support of intracranial single unit activity measurement. The parameters of the whole measuring set were tested in order to assure the optimal conditions of extracellular single-unit recording. Metal microelectrodes for measuring the single-unit were tested during animal experiments. From signals recorded during these experiments, requirements for the measuring set parameters were defined. The impedance parameters of the metal microelectrodes were measured. The frequency-gain and autonomous noise properties of preamplifier and amplifier were verified. The measurement and the description of the extracellular single unit activity could help in prognoses of brain tissue damage recovery.

Keywords: Measuring set, metal microelectrodes, single-unit, noise, impedance parameters, gain characteristics.

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Authors: R. Rajendiran, S. Gayathri devi, B.T. SureshKumar, V. Arul Priya

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Plackett-Burman statistical screening of media constituents and operational conditions for extracellular lipase production from isolate Trichoderma viride has been carried out in submerged fermentation. This statistical design is used in the early stages of experimentation to screen out unimportant factors from a large number of possible factors. This design involves screening of up to 'n-1' variables in just 'n' number of experiments. Regression coefficients and t-values were calculated by subjecting the experimental data to statistical analysis using Minitab version 15. The effects of nine process variables were studied in twelve experimental trials. Maximum lipase activity of 7.83 μmol /ml /min was obtained in the 6th trail. Pareto chart illustrates the order of significance of the variables affecting the lipase production. The present study concludes that the most significant variables affecting lipase production were found to be palm oil, yeast extract, K2HPO4, MgSO4 and CaCl2.

Keywords: lipase, submerged fermentation, statistical optimization, Trichoderma viride

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Authors: Rupinder Kaur, Parmjit S. Panesar, Ram S. Singh

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Whey is the lactose rich by-product of the dairy industry, having good amount of nutrient reservoir. Most abundant nutrients are lactose, soluble proteins, lipids and mineral salts. Disposing of whey by most of milk plants which do not have proper pre-treatment system is the major issue. As a result of which, there can be significant loss of potential food and energy source. Thus, whey has been explored as the substrate for the synthesis of different value added products such as enzymes. β-galactosidase is one of the important enzymes and has become the major focus of research due to its ability to catalyze both hydrolytic as well as transgalactosylation reaction simultaneously. The enzyme is widely used in dairy industry as it catalyzes the transformation of lactose to glucose and galactose, making it suitable for the lactose intolerant people. The enzyme is intracellular in both bacteria and yeast, whereas for molds, it has an extracellular location. The present work was carried to utilize the whey for the production of β-galactosidase enzyme using both yeast and fungal cultures. The yeast isolate Kluyveromyces marxianus WIG2 and various fungal strains have been used in the present study. Different disruption techniques have also been investigated for the extraction of the enzyme produced intracellularly from yeast cells. Among the different methods tested for the disruption of yeast cells, SDS-chloroform showed the maximum β-galactosidase activity. In case of the tested fungal cultures, Aureobasidium pullulans NCIM 1050 was observed to be the maximum extracellular enzyme producer.

Keywords: β-galactosidase, fungus, yeast, whey.

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Authors: John Justo Ambuchi, Zhaohan Zhang, Yujie Feng

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Quick development and usage of nanotechnology have resulted to massive use of various nanoparticles, such as iron oxide nanoparticles (IONPs) and multi-wall carbon nanotubes (MWCNTs). Thus, this study investigated the role of IONPs and MWCNTs in enhancing bioenergy recovery. Results show that IONPs at a concentration of 750 mg/L and MWCNTs at a concentration of 1500 mg/L induced faster substrate utilization and biogas production rates than the control. IONPs exhibited higher carbon oxygen demand (COD) removal efficiency than MWCNTs while on the contrary, MWCNT performance on biogas generation was remarkable than IONPs. Furthermore, scanning electron microscopy (SEM) investigation revealed extracellular polymeric substances (EPS) excretion from AGS had an interaction with nanoparticles. This interaction created a protective barrier to microbial consortia hence reducing their cytotoxicity. Microbial community analyses revealed genus predominance of bacteria of Anaerolineaceae and Longilinea. Their role in biodegradation of the substrate could have highly been boosted by nanoparticles. The archaea predominance of the genus level of Methanosaeta and Methanobacterium enhanced methanation process. The presence of bacteria of genus Geobacter was also reported. Their presence might have significantly contributed to direct interspecies electron transfer in the system. Exposure of AGS to nanoparticles promoted direct interspecies electron transfer among the anaerobic fermenting bacteria and their counterpart methanogens during the anaerobic digestion process. This results provide useful insightful information in understanding the response of microorganisms to IONPs and MWCNTs in the complex natural environment.

Keywords: Anaerobic granular sludge, extracellular polymeric substances, iron oxide nanoparticles, multi-wall carbon nanotubes.

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Authors: M. Saravanan

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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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Authors: Tudor Petreuş, Andrei Neamţu, Cristina Dascălu, Paul Dan Sîrbu, Carmen E. Cotrutz

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Matrix metalloproteinases (MMP) are a class of structural and functional related enzymes involved in altering the natural elements of the extracellular matrix. Most of the MMP structures are cristalographycally determined and published in WorldWide ProteinDataBank, isolated, in full structure or bound to natural or synthetic inhibitors. This study proposes an algorithm to replace missing crystallographic structures in PDB database. We have compared the results of a chosen docking algorithm with a known crystallographic structure in order to validate enzyme sites reconstruction there where crystallographic data are missing.

Keywords: matrix metalloproteinases, molecular docking, structure superposition, surface complementarity.

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Authors: Young-Seok Choi

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It has been known that a characteristic Burst-Suppression (BS) pattern appears in EEG during the early recovery period following Cardiac Arrest (CA). Here, to explore the relationship between cortical and subcortical neural activities underlying BS, extracellular activity in the parietal cortex and the centromedian nucleus of the thalamus and extradural EEG were recorded in a rodent CA model. During the BS, the cortical firing rate is extraordinarily high, and that bursts in EEG correlate to dense spikes in cortical neurons. Newly observed phenomena are that 1) thalamic activity reemerges earlier than cortical activity following CA, and 2) the correlation coefficient of cortical and thalamic activities rises during BS period. These results would help elucidate the underlying mechanism of brain recovery after CA injury.

Keywords: Cortex, thalamus, cardiac arrest, burst-suppression.

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Authors: Juliana A. Knocikova, Yann Bouret, Médéric Argentina, Laurent Counillon

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Cell volume, together with membrane potential and intracellular hydrogen ion concentration, is an essential biophysical parameter for normal cellular activity. Cell volumes can be altered by osmotically active compounds and extracellular tonicity. In this study, a simple mathematical model of osmotically induced cell swelling and shrinking is presented. Emphasis is given to water diffusion across the membrane. The mathematical description of the cellular behavior consists in a system of coupled ordinary differential equations. We compare experimental data of cell volume alterations driven by differences in osmotic pressure with mathematical simulations under hypotonic and hypertonic conditions. Implications for a future model are also discussed.

Keywords: Eukaryotic cell, mathematical modeling, osmosis, volume alterations.

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Authors: Mohammadhosein Rahimi, Mohammad Raouf Hosseini, Mehran Bakhshi, Alireza Baghbanan

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Silver ions (Ag⁺) and their compounds are consequentially toxic to microorganisms, showing biocidal effects on many species of bacteria. Silver-phosphate (or silver orthophosphate) is one of these compounds, which is famous for its antimicrobial effect and catalysis application. In the present study, a green method was presented to synthesis silver-phosphate nanoparticles using Sporosarcina pasteurii. The composition of the biosynthesized nanoparticles was identified as Ag₃PO₄ using X-ray Diffraction (XRD) and Energy Dispersive Spectroscopy (EDS). Also, Fourier Transform Infrared (FTIR) spectroscopy showed that Ag₃PO₄ nanoparticles was synthesized in the presence of biosurfactants, enzymes, and proteins. In addition, UV-Vis adsorption of the produced colloidal suspension approved the results of XRD and FTIR analyses. Finally, Transmission Electron Microscope (TEM) images indicated that the size of the nanoparticles was about 20 nm.

Keywords: Bacteria, biosynthesis, silver-phosphate, Sporosarcina pasteurii, nanoparticle.

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Authors: Ye. V. Lyubun

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The problem of agricultural-soil pollution is closely linked to the production of ecologically pure foodstuffs and to human health. An important task, therefore, is to rehabilitate agricultural soils with the help of state-of-the-art biotechnologies, based on the use of metal-accumulating plants. In this work, on the basis of literature data and the results of prior research from this laboratory, plants were selected for which the growing technology is well developed and which are widespread locally: sugar sorghum (Sorghum saccharatum), sudangrass (Sorghum sudanense (Piper.) Stapf.), and sunflower (Helianthus annuus L.). I report on laboratory experiments designed to study the influence of synthetic indole-3- acetic acid and the extracellular indole-3-acetic acid released by the plant-growth-promoting rhizobacterium Azospirillum brasilense Sp245 on growth of and arsenic accumulation by these plants.

Keywords: Arsenic, bioaccumulation, plant-growth-promoting rhizobacteria, phytohormones.

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Authors: Masoumeh Ghasemi, Afshin Farahbakhsh, Arman Farahbakhsh, Ali Asghar Safari

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In this study, lipase production has been investigated using submerge fermentation by Aspergillus niger in Kilka fish oil as main substrate. The Taguchi method with an L9 orthogonal array design was used to investigate the effect of parameters and their levels on lipase productivity. The optimum conditions for Kilka fish oil concentration, incubation temperature and pH were obtained 3 gr./ml 35°C and 7, respectively. The amount of lipase activity in optimum condition was obtained 4.59IU/ml. By comparing this amount with the amount of productivity in the olive oil medium based on the cost of each medium, it was that using Kilka fish oil is 84% economical. Therefore Kilka fish oil can be used as an economical and suitable substrate in the lipase production and industrial usages.

Keywords: Lipase, Aspergillus niger, Kilka Fish oil, Submerge Fermentation method.

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Authors: Yogesh D. Bansod, Jiri Bursa

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The quantitative study of cell mechanics is of paramount interest, since it regulates the behaviour of the living cells in response to the myriad of extracellular and intracellular mechanical stimuli. The novel experimental techniques together with robust computational approaches have given rise to new theories and models, which describe cell mechanics as combination of biomechanical and biochemical processes. This review paper encapsulates the existing continuum-based computational approaches that have been developed for interpreting the mechanical responses of living cells under different loading and boundary conditions. The salient features and drawbacks of each model are discussed from both structural and biological points of view. This discussion can contribute to the development of even more precise and realistic computational models of cell mechanics based on continuum approaches or on their combination with microstructural approaches, which in turn may provide a better understanding of mechanotransduction in living cells.

Keywords: Cell mechanics, computational models, continuum approach, mechanical models.

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Authors: A. Shyamala Devi, K. Chitra Devi, R. Rajendiran

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A total of 6 isolates of Bacillus subtilis were isolated from oil mill waste collected in Namakkal district, Tamilnadu, India. The isolated bacteria were screened using lipase screening medium containing Tween 80. BS-3 isolate exhibited a greater clear zone than the others, indicating higher lipase activity. Therefore, this isolate was selected for media optimization studies. Ten process variables were screened using Plackett–Burman design and were further optimized by central composite design of response surface methodology for lipase production in submerged fermentation. Maximum lipase production of 16.627 U/min/ml were predicted in medium containing yeast extract (9.3636g), CaCl2 (0.8986g) and incubation periods (1.813 days). A mean value of 16.98 ± 0.2286 U/min/ml of lipase was acquired from real experiments.

Keywords: Bacillus subtilis, extracellular lipase, Plackett–Burman design, response surface methodology.

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Authors: N. Kotabin, Y. Tahara, K. Issakul, O. Chunhachart

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Cadmium (II) (Cd) is one of the major toxic elemental pollutants, which is hazardous for humans, animals and plants. γ- Polyglutamic acid (γ-PGA) is an extracellular biopolymer produced by several species of Bacillus which has been reported to be an effective biosorbent for metal ions. The effect of γ-PGA on growth of rice grown under laboratory conditions was investigated. Rice seeds were germinated and then grown at 30±1°C on filter paper soaked with Cd solution and γ-PGA for 7 days. The result showed that Cd significantly inhibited the growth of roots, shoots by reducing root, and shoot lengths. Fresh and dry weights also decreased compared with control; however, the addition of 500 mg·L-1 γ-PGA alleviated rice seedlings from the adverse effects of Cd. The analysis of physiological traits revealed that Cd caused a decrease in the total chlorophyll and soluble protein contents and amylase activities in all treatments. The Cd content in seedling tissues increased for the Cd 250 μM treatment (P<0.05) but the addition of 500 mg·L-1 γ-PGA resulted in a noticeable decrease in Cd (P<0.05).

Keywords: Polyglutamic acid, Cadmium, Rice, Bacillus subtilis.

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Authors: Yulia Irnidayanti, Win Darmanto, Agus Abadi

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research goal was to determine the expression levels cDNA of brain embrio at gestation days 10 (GD-10). The Electroforesis DNA results showed that GAPDH, Fibronectin1, Ncam1, Tenascin, Vimentin, Neurofilament heavy, Neurofilament medium and Neurofilament low were 447 bp, 462 bp, 293 bp. 416 bp, 327 bp, 301 bp, 398 bp and 289 bp. Result of real-time RT-PCR on brain Embryo at gestation days 10 showed that the expression of copy gen Fibronectin 36 copies, Ncam 21,708 copies; Tenascin 24,505 copies; Vimentin 538,554 copies; Neurofilament heavy 2,419 copies; Neurofilament medium 92,928 copies; Neurofilament low 125,809 copies. Vimentin expressed gene copies is very high compared with other gene copies. This condition are caused by Vimentin, that contribute to proliferate of brain development. The vimentin role to cell proliferation of brain.

Keywords: GAPDH, Fibronectin, Ncam, Tenascin, vimentin, Neurofilamen heavy, Neurofilament medium, Neurofilamen low.

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Authors: Deepak Mohan, Sushma Sharma

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Diclofenac sodium, a member of the acetic acid family of non-steroidal anti-inflammatory drugs, is used to retard inflammation, arthritis pain and ankylosing spondylitis. The drug is known to cause severe injury in different tissues due to formation of reactive oxygen species. The present study is focused on the effect of different doses of diclofenac (4 mg/kg/body weight and 14 mg/kg/body weight on histoarchitecture of the liver from 7-28 days of the investigation. Diclofenac administration resulted in distorted hepatic degeneration and formation of wide areas in the form of sinusoidal gaps. Hepatic fibrosis noticed in different stages of investigation could be attributed to chronic inflammation and reactive oxygen species which results in deposition of extracellular matrix proteins. The abrupt degenerative changes observed during later stages of the experiment showed maximum damage to the liver, and there was enlargement of sinusoidal gaps accompanied by maximum necrosis in the tissues.

Keywords: Arthritis, diclofenac, histoarchitecture, sinusoidal.

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Authors: Le Thuy Mai, Vu Nguyen Thanh

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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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Authors: Noor Muzamil Mohamad, Abdul Manaf Ali, Hamzah Mohd Salleh

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Phytases (myo-inositol hexakisphosphate phosphohydrolases; EC 3.1.3.8) catalyze the hydrolysis of phytic acid (myoinositol hexakisphosphate) to the mono-, di-, tri-, tetra-, and pentaphosphates of myo-inositol and inorganic phosphate. Therrmophilic bacteria isolated from water sampled from hot spring. About 120 isolates of bacteria were successfully isolated form hot spring water sample and tested for extracellular phytase producing. After 5 passages of the screening on the PSM media, 4 isolates were found stable in producing phytase enzyme. The 16s RDNA sequencing for identification of bacteria using molecular technique revealed that all isolates those positive in phytase producing are belong to Geobacillus spp. And Anoxybacillus spp. Anoxybacillus rupiensis UniSZA-7 were identified for their carbon source utilization using Phenotype Microarray Plate of Biolog and found they utilize several kind of carbon source provided.

Keywords: Phytase, Phytic Acid, Thermophilic Bacteria, PSM Media and Phytase Assay

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Authors: M. Maimunah, G. A. Froemming, H. Nawawi, M. I. Nafeeza, O. Effat, M. Y. Rosmadi, M. S. Mohamed Saifulaman

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Atherosclerosis was identified as a chronic inflammatory process resulting from interactions between plasma lipoproteins, cellular components (monocyte, macrophages, T lymphocytes, endothelial cells and smooth muscle cells) and the extracellular matrix of the arterial wall. Several types of genes were known to express during formation of atherosclerosis. This study is carried out to identify unknown differentially expressed gene (DEG) in atherogenesis. Rabbit’s aorta tissues were stained by H&E for histomorphology. GeneFishing™ PCR analysis was performed from total RNA extracted from the aorta tissues. The DNA fragment from DEG was cloned, sequenced and validated by Real-time PCR. Histomorphology showed intimal thickening in the aorta. DEG detected from ACP-41 was identified as cathepsin B gene and showed upregulation at week-8 and week-12 of atherogenesis. Therefore, ACP-based GeneFishing™ PCR facilitated identification of cathepsin B gene which was differentially expressed during development of atherosclerosis.

Keywords: Atherosclerosis, GeneFishing™ PCR, cathepsin B gene.

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Authors: Forough Ataollahi, Belinda Pingguan-Murphy, Wan Abu Bakar Wan Abas, Noor Azuan Abu Osman

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Endothelium proliferation is an important process in cardiovascular homeostasis and can be regulated by extracellular environment, as cells can actively sense mechanical environment. In this study, we evaluated endothelial cell proliferation on PDMS/alumina (Al₂O₃) composites and pure PDMS. The substrates were prepared from pure PDMS and its composites with 5% and 10% Al₂O₃ at curing temperature 50˚C for 4h and then characterized by mechanical, structural and morphological analyses. Higher stiffness was found in the composites compared to the pure PDMS substrate. Cell proliferation of the cultured bovine aortic endothelial cells on substrate materials were evaluated via Resazurin assay and 1, 1’-Dioctadecyl-1, 3, 3, 3’, 3’-Tetramethylindocarbocyanine Perchlorate-Acetylated LDL (Dil-Ac-LDL) cell staining, respectively. The results revealed that stiffer substrates promote more endothelial cells proliferation to the less stiff substrates. Therefore, this study firmly hypothesizes that the stiffness elevates endothelial cells proliferation.

Keywords: Bovine aortic endothelial cells, extra cellular matrix, proliferation, stiffness.

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Authors: R. S. A. Nesbitt, J. Macione, E. Babollah, B. Adu-baffour, S. P. Kotha

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F-actin fibrils are the cytoskeleton of osteocytes. They react in a dynamic manner to mechanical loading, and strength and reposition their efforts to reinforce the cells structure. We hypothesize that f-actin is temporarly disrupted after loading and repolymerizes in a new orientation to oppose the applied load. In vitro studies are conducted to determine f-actin disruption after varying mechanical stimulus parameters that are known to affect bone formation. Results indicate that the f-actin cytoskeleton is disrupted in vitro as a function of applied mechanical stimulus parameters and that the f-actin bundles reassemble after loading induced disruption within 3 minutes after cessation of loading. The disruption of the factin cytoskeleton depends on the magnitude of stretch, the numbers of loading cycles, frequency, the insertion of rest between loading cycles and extracellular calcium. In vivo studies also demonstrate disruption of the f-actin cytoskeleton in cells embedded in the bone matrix immediately after mechanical loading. These studies suggest that adaptation of the f-actin fiber bundles of the cytoskeleton in response to applied loads occurs by disruption and subsequent repolymerization.

Keywords: Mechanical loading of osteocytes, f-actin cytoskeleton, disruption, re-polymerization.

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Authors: M. F. R. Zuthi, H. H. Ngo, W. S. Guo, S. S. Chen, N. C. Nguyen, L. J. Deng, T. D. C. Tran

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Sustaining a desired rate of oxygen transfer for microbial activity is a matter of major concern for biological wastewater treatment (MBR). The study reported in the paper was aimed at assessing the effects of microbial products on the specific oxygen uptake rate (SOUR) in a conventional membrane bioreactor (CMBR) and that in a sponge submerged MBR (SSMBR). The production and progressive accumulation of soluble microbial products (SMP) and bound-extracellular polymeric substances (bEPS) were affecting the SOUR of the microorganisms which varied at different stages of operation of the MBR systems depending on the variable concentrations of the SMP/bEPS. The effect of bEPS on the SOUR was stronger in the SSMBR compared to that of the SMP, while relative high concentrations of SMP had adverse effects on the SOUR of the CMBR system. Of the different mathematical correlations analyzed in the study, logarithmic mathematical correlations could be established between SOUR and bEPS in SSMBR, and similar correlations could also be found between SOUR and SMP concentrations in the CMBR.

Keywords: Microbial products, Microbial activity, Specific oxygen uptake rate, Membrane bioreactor.

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Authors: S.H. Mirdamadian, S.M. Khayam-Nekoui, H. Ghanavati

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Composting is the process in which municipal solid waste (MSW) and other organic waste materials such as biosolids and manures are decomposed through the action of bacteria and other microorganisms into a stable granular material which, applied to land, as soil conditioner. Microorganisms, especially those that are able to degrade polymeric organic material have a key role in speed up this process. The aim of this study has been established to isolation of microorganisms with high ability to production extracellular enzymes for degradation of natural polymers that are exists in MSW for decreasing time of degradation phase. Our experimental study for isolation designed in two phases: in first phase we isolated degrading microorganism with selected media that consist a special natural polymer such as cellulose, starch, lipids and etc as sole source of carbon. In second phase we selected microorganism that had high degrading enzyme production with enzymatic assay for seed production. However, our findings in pilot scale have indicated that usage of this microbial consortium had high efficiency for decreasing degradation phase.

Keywords: Biodegradation, Compost, Municipal Solid Waste, Waste Management.

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Authors: Lina Paola Orozco-Marín, Yuliet Montoya, John Bustamante

Abstract:

Ischemic events can culminate in acute myocardial infarction with irreversible cardiac lesions that cannot be restored due to the limited regenerative capacity of the heart. Tissue engineering proposes therapeutic alternatives by using biomaterials to resemble the native extracellular medium combined with healthy and functional cells. This research focused on developing a natural thermosensitive hydrogel, its physical-chemical characterization and in vitro biocompatibility determination. Hydrogels’ morphological characterization was carried out through scanning electron microscopy and its chemical characterization by employing Infrared Spectroscopy technic. In addition, the biocompatibility was determined using fetal human ventricular cardiomyocytes cell line RL-14 and the MTT cytotoxicity test according to the ISO 10993-5 standard. Four biocompatible and thermosensitive hydrogels were obtained with a three-dimensional internal structure and two gelation times. The results show the potential of the hydrogel to increase the cell survival rate to the cardiac cell therapies under investigation and lay the foundations to continue with its characterization and biological evaluation both in vitro and in vivo models.

Keywords: cardiac cell therapy, cardiac ischemia, natural polymers, thermosensitive hydrogel

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Authors: Osama M. Darwesh, Sahar H. Hassan, Abd El-Raheem R. El-Shanshoury, Shawky Z. Sabae

Abstract:

Nanotechnology as multidisciplinary technology is growing rapidly with important applications in several sectors. Also, nanobiotechnology is known for the use of microorganisms for the synthesis of targeted nanoparticles. The present study deals with the green synthesis of silver nanoparticles using aquatic bacteria and the development of a biogenic nanocomposite for environmental applications. 20 morphologically different colonies were isolated from the collected water samples from eight different locations at the Rosetta branch of the Nile Delta, Egypt. The obtained results illustrated that the most effective bacterial isolate (produced the higher amount of AgNPs after 24 h of incubation time) is isolate R3. Bacillus tequilensis was the strongest extracellular bio-manufactory of AgNPs. Biosynthesized nanoparticles had a spherical shape with a mean diameter of 2.74 to 28.4 nm. The antimicrobial activity of silver nanoparticles against many pathogenic microbes indicated that the produced AgNPs had high activity against all tested multi-antibiotic resistant pathogens. Also, the stabilized prepared AgNPs-SA nanocomposite has greater catalytic activity for the decolourization of some dyes like Methylene blue (MB) and Crystal violet. Such results represent a promising stage for producing eco-friendly, cost-effective, and easy-to-handle devices for the bioremediation of contaminated industrial wastewater.

Keywords: Bioremediation, AgNPs, AgNPs-SA nanocomposite, Bacillus tequilensis, nanobiotechnology.

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