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

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

The Effect of Carboxymethyl Cellulose on the Stability of Emulsions Stabilized by Whey Proteins under Digestion in vitro and in vivo

In vitro gastro-duodenal digestion model was used to investigate the changes of emulsions under digestion conditions. Oil in water emulsions stabilized by whey proteins (2%) and stabilized by whey proteins (2%) with addition of carboxymethyl cellulose (0.75%) as gelling agent of continuous phase were prepared at pH7. Both emulsions were destabilized under gastric conditions; however the protective role of carboxymethyl cellulose was indicated by recording delay of fat digestibility of this emulsion. In the presence of carboxymethyl cellulose whey proteins on the interfacial surface of droplets were more resistant to gastric degradation causing limited hydrolysis of fat due to the poor acceptability of lipids for the enzymes. Studies of emulsions using in vivo model supported results from in vitro studies. Lower content of triglycerides in blood serum and higher amount of fecal fat of rats were determined when rats were fed by diet containing emulsion made with whey proteins and carboxymethyl cellulose. 

Double Immobilized Lipase for the Kinetic Resolution of Secondary Alcohols

Sol-gel immobilization of enzymes, which can improve considerably their properties, is now one of the most used techniques. By deposition of the entrapped lipase on a solid support, a new and improved biocatalyst was obtained, which can be used with excellent results in acylation reactions. In this paper, lipase B from Candida antarctica was double immobilized on different adsorbents. These biocatalysts were employed in the kinetic resolution of several aliphatic secondary alcohols in organic medium. High total recovery yields of enzymatic activity, up to 560%, were obtained. For all the studied alcohols the enantiomeric ratios E were over 200. The influence of the reaction medium was studied for the kinetic resolution of 2-pentanol.

Molecular Docking on Recomposed versus Crystallographic Structures of Zn-Dependent Enzymes and their Natural Inhibitors

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.

Estimating Enzyme Kinetic Parameters from Apparent KMs and Vmaxs

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

Mutational Effect to Particular Interaction Energy of Cycloguanil Drug to Plasmodium Plasmodium Falciparum Dihydrofolate Reductase Enzymes

In order to find the particular interaction energy between cylcloguanil and the amino acids surrounding the pocket of wild type and quadruple mutant type PfDHFR enzymes, the MP2 method with basis set 6-31G(d,p) level of calculations was performed. The obtained interaction energies found that Asp54 has the strongest interaction energy to both wild type and mutant type of - 12.439 and -11.250 kcal/mol, respectively and three amino acids; Asp54, Ile164 and Ile14 formed the H-bonding with cycloguanil drug. Importantly, the mutation at Ser108Asn was the key important of cycloguanil resistant with showing repulsive interaction energy.

Immobilization of Aspergillus awamori 1-8 for Subsequent Pectinase Production

The overall objective of this research is a strain improvement technology for efficient pectinase production. A novel cells cultivation technology by immobilization of fungal cells has been studied in long time continuous fermentations. Immobilization was achieved by using of new material for absorption of stores of immobilized cultures which was for the first time used for immobilization of microorganisms. Effects of various conditions of nitrogen and carbon nutrition on the biosynthesis of pectolytic enzymes in Aspergillus awamori 1-8 strain were studied. Proposed cultivation technology along with optimization of media components for pectinase overproduction led to increased pectinase productivity in Aspergillus awamori 1-8 from 7 to 8 times. Proposed technology can be applied successfully for production of major industrial enzymes such as α-amylase, protease, collagenase etc.

All Proteins Have a Basic Molecular Formula

This study proposes a basic molecular formula for all proteins. A total of 10,739 proteins belonging to 9 different protein groups classified on the basis of their functions were selected randomly. They included enzymes, storage proteins, hormones, signalling proteins, structural proteins, transport proteins, immunoglobulins or antibodies, motor proteins and receptor proteins. After obtaining the protein molecular formula using the ProtParam tool, the H/C, N/C, O/C, and S/C ratios were determined for each randomly selected sample. In this case, H, N, O, and S coefficients were specified per carbon atom. Surprisingly, the results demonstrated that H, N, O, and S coefficients for all 10,739 proteins are similar and highly correlated. This study demonstrates that despite differences in the structure and function, all known proteins have a similar basic molecular formula CnH1.58 ± 0.015nN0.28 ± 0.005nO0.30 ± 0.007nS0.01 ± 0.002n. The total correlation between all coefficients was found to be 0.9999.

Changes to Oxidative Stress Levels Following Exposure to Formaldehyde in Lymphocytes

Formaldehyde is the illegal chemical substance used for food preservation in fish and vegetable. It can promote carcinogenesis. Superoxide dismutases are the important antioxidative enzymes that catalyze the dismutation of superoxide anion into oxygen and hydrogen peroxide. The resultant level of oxidative stress in formaldehyde-treated lymphocytes was investigated. The formaldehyde concentrations of 0, 20, 40, 60, 80 and 120μmol/L were treated in human lymphocytes for 12 hours. After 12 treated hours, the superoxide dismutase activity change was measured in formaldehyde-treated lymphocytes. The results showed that the formaldehyde concentrations of 60, 80 and 120μmol/L significantly decreased superoxide dismutase activities in lymphocytes (P < 0.05). The change of superoxide dismutase activity in formaldehyde-treated lymphocytes may be the biomarker for detect cellular injury, such as damage to DNA, due to formaldehyde exposure.

A Novel Strategy for Oriented Protein Immobilization

A new strategy for oriented immobilization of proteins was proposed. The strategy contains two steps. The first step is to search for a docking site away from the active site on the protein surface. The second step is trying to find a ligand that is able to grasp the targeted site of the protein. To avoid ligand binding to the active site of protein, the targeted docking site is selected to own opposite charges to those near the active site. To enhance the ligand-protein binding, both hydrophobic and electrostatic interactions need to be included. The targeted docking site should therefore contain hydrophobic amino acids. The ligand is then selected through the help of molecular docking simulations. The enzyme α-amylase derived from Aspergillus oryzae (TAKA) was taken as an example for oriented immobilization. The active site of TAKA is surrounded by negatively charged amino acids. All the possible hydrophobic sites on the surface of TAKA were evaluated by the free energy estimation through benzene docking. A hydrophobic site on the opposite side of TAKA-s active site was found to be positive in net charges. A possible ligand, 3,3-,4,4- – Biphenyltetra- carboxylic acid (BPTA), was found to catch TAKA by the designated docking site. Then, the BPTA molecules were grafted onto silica gels and measured the affinity of TAKA adsorption and the specific activity of thereby immobilized enzymes. It was found that TAKA had a dissociation constant as low as 7.0×10-6 M toward the ligand BPTA on silica gel. The increase in ionic strength has little effect on the adsorption of TAKA, which indicated the existence of hydrophobic interaction between ligands and proteins. The specific activity of the immobilized TAKA was compared with the randomly adsorbed TAKA on primary amine containing silica gel. It was found that the orderly immobilized TAKA owns a specific activity twice as high as the one randomly adsorbed by ionic interaction.

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

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

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

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

Optimization of Enzymatic Activities in Malting of Oat

Malting is usually carried out on intact barley seed, while hull is still attached to it. In this study, oat grain with and without hull was subjected to controlled germination to optimize its enzymes activity, in such a way that lipase has the lowest and α- amylase and proteinase the highest activities. Since pH has a great impact on the activity of the enzymes, the pH of germination media was set up to 3 to 8. In dehulled oats, lipase and α-amylase had the lowest and highest activities in pHs 3 and 6, respectively whereas the highest proteinase activity was evidenced at pH 7 and 4 in the oats with and without hull respectively. While measurements indicated that the effect of hull on the enzyme activities particularly in lipase and amylase at each level of the pH are significantly different, the best results were obtained in those samples in which their hull had been removed. However, since the similar lipase activity in germinated dehulled oat were recorded at the pHs 4 and 5, therefore it was concluded that pH 5 in dehulled oat seed may provide the optimum enzyme activity for all the enzymes.

Alcoholic Extract of Terminalia Arjuna Protects Rabbit Heart against Ischemic-Reperfusion Injury: Role of Antioxidant Enzymes and Heat Shock Protein

The present study was designed to investigate the cardio protective role of chronic oral administration of alcoholic extract of Terminalia arjuna in in-vivo ischemic reperfusion injury and the induction of HSP72. Rabbits, divided into three groups, and were administered with the alcoholic extract of the bark powder of Terminalia arjuna (TAAE) by oral gavage [6.75mg/kg: (T1) and 9.75mg/kg: (T2), 6 days /week for 12 weeks]. In open-chest Ketamine pentobarbitone anaesthetized rabbits, the left anterior descending coronary artery was occluded for 15 min of ischemia followed by 60 min of reperfusion. In the vehicle-treated group, ischemic-reperfusion injury (IRI) was evidenced by depression of global hemodynamic function (MAP, HR, LVEDP, peak LV (+) & (- ) (dP/dt) along with depletion of HEP compounds. Oxidative stress in IRI was evidenced by, raised levels of myocardial TBARS and depletion of endogenous myocardial antioxidants GSH, SOD and catalase. Western blot analysis showed a single band corresponding to 72 kDa in homogenates of hearts from rabbits treated with both the doses. In the alcoholic extract of the bark powder of Terminalia arjuna treatment groups, both the doses had better recovery of myocardial hemodynamic function, with significant reduction in TBARS, and rise in SOD, GSH, catalase were observed. The results of the present study suggest that the alcoholic extract of the bark powder of Terminalia arjuna in rabbit induces myocardial HSP 72 and augments myocardial endogenous antioxidants, without causing any cellular injury and offered better cardioprotection against oxidative stress associated with myocardial IR injury.

Construction of Recombinant E.coli Expressing Fusion Protein to Produce 1,3-Propanediol

In this study, a synthetic pathway was created by assembling genes from Clostridium butyricum and Escherichia coli in different combinations. Among the genes were dhaB1 and dhaB2 from C. butyricum VPI1718 coding for glycerol dehydratase (GDHt) and its activator (GDHtAc), respectively, involved in the conversion of glycerol to 3-hydroxypropionaldehyde (3-HPA). The yqhD gene from E.coli BL21 was also included which codes for an NADPHdependent 1,3-propanediol oxidoreductase isoenzyme (PDORI) reducing 3-HPA to 1,3-propanediol (1,3-PD). Molecular modeling analysis indicated that the conformation of fusion protein of YQHD and DHAB1 was favorable for direct molecular channeling of the intermediate 3-HPA. According to the simulation results, the yqhD and dhaB1 gene were assembled in the upstream of dhaB2 to express a fusion protein, yielding the recombinant strain E. coliBL21 (DE3)//pET22b+::yqhD-dhaB1_dhaB2 (strain BP41Y3). Strain BP41Y3 gave 10-fold higher 1,3-PD concentration than E. coliBL21 (DE3)//pET22b+::yqhD-dhaB1_dhaB2 (strain BP31Y2) expressing the recombinant enzymes simultaneously but in a non-fusion mode. This is the first report using a gene fusion approach to enhance the biological conversion of glycerol to the value added compound 1,3- PD.

Characterization of Corn Cobs from Microwave and Potassium Hydroxide Pretreatment

The complexity of lignocellulosic biomass requires a pretreatment step to improve the yield of fermentable sugars. The efficient pretreatment of corn cobs using microwave and potassium hydroxide and enzymatic hydrolysis was investigated. The objective of this work was to characterize the optimal condition of pretreatment of corn cobs using microwave and potassium hydroxide enhance enzymatic hydrolysis. Corn cobs were submerged in different potassium hydroxide concentration at varies temperature and resident time. The pretreated corn cobs were hydrolyzed to produce the reducing sugar for analysis. The morphology and microstructure of samples were investigated by Thermal gravimetric analysis (TGA, scanning electron microscope (SEM), X-ray diffraction (XRD). The results showed that lignin and hemicellulose were removed by microwave/potassium hydroxide pretreatment. The crystallinity of the pretreated corn cobs was higher than the untreated. This method was compared with autoclave and conventional heating method. The results indicated that microwave-alkali treatment was an efficient way to improve the enzymatic hydrolysis rate by increasing its accessibility hydrolysis enzymes.

Effect of Crude oil Intoxication on Antioxidant and Marker Enzymes of Tissue Damage in Liver of Rat

The objective of the present study was to examine the dose-response relationships between antioxidant parameters and liver contaminant levels of Kazakhstan light crude oil (KLCO) in albino rats. The animals were repeatedly exposed, by intraperitoneal injection, to low dosages (0.5–1.5 ml/kg) of KLCO. Rats exposed to these doses levels did not show any apparent symptoms of intoxication. Serum aminotransferases increased significantly (p

The Effect of Selective Cyclooxygenase (COX) Inhibitors on Japanese Medaka (Oryzias latipes) Reproduction Parameters

Our results showed that treatment with both cyclooxygenase (COX1 or COX2) inhibitors impair reproduction parameters of the medaka. Resveratrol (COX1 inhibitor) caused an decrease in the number of spawning females at the first week of feeding fish with experimental diets. In the group treated with NS- 398 (COX2 inhibitor) we found the lowest sperm velocity parameters and decreased linearity of movement. The ovaries of the medaka fed feed supplemented with Resveratrol or NS-398 were confirmed to have a lower share of matured oocytes however during the experiment (four weeks) the number of eggs spawned by females was similar. Both inhibitors in fish diet (20 mg/kg body weight/day) caused a decrease in the embryo survival. Our results revealed that for the medaka female reproduction, activity of both COX enzymes might be necessary whereas males reproduction competence, as expressed by sperm motility parameters, might be related to COX2 activity.

Enhancement of Essential Oil from Agarwood by Subcritical Water Extraction and Pretreatments on Hydrodistillation

The traditional method for essential oil extraction from agarwood (Aquilaria Crassna) is to soak it in water and follow with hydrodistillation. The effect of various agarwood pretreatments: ethanol, acid, alkaline, enzymes, and ultrasound, and the effect of subcritical water extraction(SWE) was studied to compare with the traditional method. The major compositions of agarwood oil from hydrodistillation were aroma compounds as follow: aristol-9-en-8- one (21.53%), selina-3, 7(11)-diene (12.96%), τ-himachalene (9.28%), β-guaiene (5.79%), hexadecanoic acid (4.90%) and guaia- 3,9-diene (4.21%). Whereas agarwood oil from pretreatments with ethanol and ultrasound, and SWE got fatty acid compounds. Extraction of agarwood oil using these pretreatments could improve the agarwood oil yields up to 2 times that of the traditional method. The components of the pretreated sample with diluted acid (H2SO4) at pH 4 gave quite similar results as the traditional method. Therefore, the enhancement of essential oil from agarwood depends on requirement of type of extracted oil that involved extraction methods.

Mechanisms of Organic Contaminants Uptake and Degradation in Plants

As a result of urbanization, the unpredictable growth of industry and transport, production of chemicals, military activities, etc. the concentration of anthropogenic toxicants spread in nature exceeds all the permissible standards. Most dangerous among these contaminants are organic compounds having great persistence, bioaccumulation, and toxicity along with our awareness of their prominent occurrence in the environment and food chain. Among natural ecological tools, plants still occupying above 40% of the world land, until recently, were considered as organisms having only a limited ecological potential, accumulating in plant biomass and partially volatilizing contaminants of different structure. However, analysis of experimental data of the last two decades revealed the essential role of plants in environment remediation due to ability to carry out intracellular degradation processes leading to partial or complete decomposition of carbon skeleton of different structure contaminants. Though, phytoremediation technologies still are in research and development, their various applications have been successfully used. The paper aims to analyze mechanisms of organic contaminants uptake and detoxification in plants, being the less studied issue in evaluation and exploration of plants potential for environment remediation.