Search results for: gastrointestinal enzymes

Authors: D.T. Mirzarakhmetova

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The studying of enzymatic esterification of carboxylic acids and higher alcohols was performed by esterase Saccharomyces cerevisiae in water-organic medium. Investigation of the enzyme specificity to acetic substrates showed the best result with acetic acid in esterification reactions with ethanol whereas within other carboxylic acids the esterification decreased with acids: hexanoic > pentanoic > butyric > decanoic. In relation to higher alcohols C3-C5, esterification increased with alcohols propanol < butanol < amylol. Also it was determined that esterase was more specific to alcohols with branched chain such as isobutyl alcohol and isoamyl alcohol. Data obtained may have important practical implications, for example, for application of yeast esterase in producing various volatile esters as well as in enzymatic transformation of volatile acids and toxic fusel alcohols into volatile esters by providing the production of the high quality alcoholic beverages with redused content of higher alcohols as well as with improved degustational and hygienic properties.

Keywords: enzymes in non-conventional media, esterification, higher alcohols, volatile esters, yeast esterase

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Authors: Nuttawan Yoswathana, M. N. Eshiaghi, K. Jaturapornpanich

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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.

Keywords: Agarwood, aquilaria crassna, hydrodistillation, subcritical water extraction.

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Authors: R. U. Hamzah, H. L. Muhammad, A. Sayyadi, M. B Busari, R. Garba, M. B. Umar, A. N Abubakar

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Lead (Pb) and Cadmium (Cd) are toxic, non-essential transition metals that pose many health risks for both humans and animals. They are environmental toxicants which contribute to testicular damage resulting to infertility problem among male populace worldwide. Chelating agents used for lead and cadmium toxicity are not readily available, toxic, expensive and unable to mop up most of the toxic metals accumulated in various organs. In this study, the effect of crude extract (CE), ethyl acetate fraction (EF) and acetone fraction (AF) of Pterocarpus mildbraedii leaf extract was assessed on cadmium-lead chloride induced testicular damaged in male albino Wistar rats. CE of the leaf was obtained by extracting in absolute methanol which was further subjected to solvent partitioning via vacuum liquid chromatographic (VLC) techniques using ethyl acetate, acetone and 70% methanol. A preliminary phytochemical screening and in vitro antioxidants guided activities on the CE and fractions were determined using standard methods. EF, AF and CE which exhibited significant in vitro activity were subjected to an in vivo study using Wistar rats. In vivo antioxidant markers, male reproductive hormones, testicular enzymes and DNA damage markers were analyzed on the rats’ testes supernatant. AF had the highest quantities of phenols (319.00 mg/g), flavonoids (8.87 mg/g) and tannins (8.87 mg/g) while methanol and EFs were richer in saponins (135.32 µg/g) and alkaloids (38.34 µg/g) respectively. A dose dependent 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, ferric reducing antioxidant power (FRAP) and lipid peroxidation were observed in all the extract with high antioxidants power in CE and AF. Administration of lead-cadmium chloride solution significantly (p > 0.05) decreases the testicular superoxide dismutase (SOD) activity to 6.82 unit/mg protein, Catalase (CAT) activity to 8.07 of H2O2 consumed/unit/mg protein and Glutathione (GSH) concentration to 31.30 ug/mg protein. There was a concomitant increase in the level of Malondialdehyde (MDA) to a value of 23.70 mmol/mg protein. In addition, lead-cadmium chloride solution significantly (p > 0.05) increases the testicular marker enzymes (Alkaline phosphatase (119.57 u/L), lactate dehydrogenase (357.05 u/L), Acid phosphatase (98.65 u/L)) and DNA damage markers (conjugated dienes (93.39 nmol/mg protein), carbonyl protein (35.39 nmol/mg protein), DNA fragmentation percentage (32.12%)) with lowered testicular hormones (Testosterone (3.1 ng/mL), Follicle stimulating (0.35 IU/mL) and Luteinizing hormones (0.15 IU/mL)) of the animals in negative control group when compared with other treated groups. Treatment with Pterocarpus mildbraedii leaf extract reverts the observed changes with the best activities found in the CE and AFs in a dose dependent manner. Pterocarpus mildbraedii leaf extract ameliorated the lead/cadmium induced testicular damage in male albino rats. The restoration of the aforementioned parameters by some of the extract dosages were comparable to the standard drug with higher activities in the crude and AF. Therefore, Pterocarpus mildbraedii leaf extract can be explored further for the management of lead/cadmium induced toxicity.

Keywords: Cadmium, lead, Pterocarpus mildbraedii, testicular damage.

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Authors: Prachi Singh

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This paper presents a low-cost, eco-friendly and reproducible microbe mediated biosynthesis of TiO₂ nanoparticles. TiO₂ nanoparticles synthesized using the bacterium, Bacillus subtilis, from titanium as a precursor, were confirmed by TEM analysis. The morphological characteristics state spherical shape, with the size of individual or aggregate nanoparticles, around 30-40 nm. Microbial resistance represents a challenge for the scientific community to develop new bioactive compounds. Here, the antibacterial effect of TiO₂ nanoparticles on Escherichia coli was investigated, which was confirmed by CFU (Colony-forming unit). Further, growth curve study of E. coli Hb101 in the presence and absence of TiO₂ nanoparticles was done. Optical density decrease was observed with the increase in the concentration of TiO₂. It could be attributed to the inactivation of cellular enzymes and DNA by binding to electron-donating groups such as carboxylates, amides, indoles, hydroxyls, thiols, etc. which cause little pores in bacterial cell walls, leading to increased permeability and cell death. This justifies that TiO₂ nanoparticles have efficient antibacterial effect and have potential to be used as an antibacterial agent for different purposes.

Keywords: Antibacterial effect, CFU, Escherichia coli Hb101, growth curve, TEM, TiO2 nanoparticle, toxicity, UV-Vis.

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

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

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

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Authors: Boonyisa Wanitwattanarumlug, Apanee Luengnaruemitchai, Sujitra Wongkasemjit

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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.

Keywords: Corn cobs, Enzymatic hydrolysis, Microwave, Potassium hydroxide, Pretreatment.

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Authors: D. Leskauskaite, I. Jasutiene, M. Kersiene, E. Malinauskyte, P. Matusevicius

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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. 

Keywords: Digestibility, emulsions, lipids, rats.

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

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

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

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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: Rotchanaphan Hengaroonprasan, Malinee Sriariyanun, Prapakorn Tantayotai, Supacharee Roddecha, Kraipat Cheenkachorn

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Lignocellolusic material is a substance that is resistant to be degraded by microorganisms or hydrolysis enzymes. To be used as materials for biofuel production, it needs pretreatment process to improve efficiency of hydrolysis. In this work, chemical pretreatments on rice straw using three diluted organic acids, including acetic acid, citric acid, oxalic acid, were optimized. Using Response Surface Methodology (RSM), the effect of three pretreatment parameters, acid concentration, treatment time, and reaction temperature, on pretreatment efficiency were statistically evaluated. The results indicated that dilute oxalic acid pretreatment led to the highest enhancement of enzymatic saccharification by commercial cellulase and yielded sugar up to 10.67 mg/ml when using 5.04% oxalic acid at 137.11 oC for 30.01 min. Compared to other acid pretreatment by acetic acid, citric acid, and hydrochloric acid, the maximum sugar yields are 7.07, 6.30, and 8.53 mg/ml, respectively. Here, it was demonstrated that organic acids can be used for pretreatment of lignocellulosic materials to enhance of hydrolysis process, which could be integrated to other applications for various biorefinery processes. 

Keywords: Lignocellolusic biomass, pretreatment, organic acid response surface methodology, biorefinery.

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Authors: Anna Yu. Muratova, Vera V. Kapitonova, Marina P. Chernyshova, Olga V. Turkovskaya

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This research was undertaken to study enzymatic activity in the shoots, roots, and rhizosphere of alfalfa (Medicago sativa L.) grown in quartz sand that was uncontaminated and contaminated with phenanthrene at concentrations of 10 and 100 mg kg-1. The higher concentration of phehanthrene had a distinct phytotoxic effect on alfalfa, inhibiting seed germination energy, plant survival, and biomass accumulation. The plant stress response to the environmental pollution was an increase in peroxidase activity. Peroxidases were the predominant enzymes in the alfalfa shoots and roots. The peroxidase profile in the shoots differed from that in the roots and had different isoenzyme numbers. 2,2'-Azinobis-(3-ethylbenzo-thiazoline-6-sulphonate) (ABTS) peroxidase was predominant in the shoots, and 2,7-diaminofluorene (2,2-DAF) peroxidase was predominant in the roots. Under the influence of phenanthrene, the activity of 2,7-DAF peroxidase increased in the shoots, and the activity of ABTS peroxidase increased in the roots. Alfalfa root peroxidases were the prevalent enzyme systems in the rhizosphere sand. Examination of the activity of alfalfa root peroxidase toward phenanthrene revealed the possibility of involvement of the plant enzyme in rhizosphere degradation of the PAH.

Keywords: Medicago sativa, enzymatic activity, peroxidase, phenanthrene.

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Authors: C. Zarcula, R. Croitoru, L. Corîci, C. Csunderlik, F. Peter

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Lipases are enzymes particularly amenable for immobilization by entrapment methods, as they can work equally well in aqueous or non-conventional media and long-time stability of enzyme activity and enantioselectivity is needed to elaborate more efficient bioprocesses. The improvement of Pseudomonas fluorescens (Amano AK) lipase characteristics was investigated by optimizing the immobilization procedure in hybrid organic-inorganic matrices using ionic liquids as additives. Ionic liquids containing a more hydrophobic alkyl group in the cationic moiety are beneficial for the activity of immobilized lipase. Silanes with alkyl- or aryl nonhydrolizable groups used as precursors in combination with tetramethoxysilane could generate composites with higher enantioselectivity compared to the native enzyme in acylation reactions of secondary alcohols. The optimal effect on both activity and enantioselectivity was achieved for the composite made from octyltrimethoxysilane and tetramethoxysilane at 1:1 molar ratio (60% increase of total activity following immobilization and enantiomeric ratio of 30). Ionic liquids also demonstrated valuable properties as reaction media for the studied reactions, comparable with the usual organic solvent, hexane.

Keywords: Ionic liquids, lipase, enantioselectivity, sol-gelimmobilization

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Authors: R. Majidzadeh Heravi, M. Sankian, H. Kermanshahi, M. R. Nassiri, A. Heravi Moussavi, S. A. Lari, A. R. Varasteh

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The use of probiotics engineered to express specific enzymes has been the subject of considerable attention in poultry industry because of increased nutrient availability and reduced cost of enzyme supplementation. Phytase enzyme is commonly added to poultry feed to improve digestibility and availability of phosphorus from plant sources. To construct a probiotic with potential of phytate degradation, phytase gene (appA) from E. coli was cloned and transformed into two probiotic bacteria Lactobacillus salivarius and Lactococcus lactis. L. salivarous showed plasmid instability, unable to express the gene. The expression of appA gene in L. lactis was analyzed by detecting specific RNA and zymography assay. Phytase enzyme was isolated from cellular extracts of recombinant L. lactis, showing a 46 kDa band upon the SDS-PAGE analysis. Zymogram also confirmed the phytase activity of the 46 kDa band corresponding to the enzyme. An enzyme activity of 4.9U/ml was obtained in cell extracts of L. lactis. The growth of native and recombinant L. lactis was similar in the presence of two concentrations of ox bile.

Keywords: Lactobacillus salivarus, Lactococcus lactis, recombinant, phytase, poultry.

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Authors: Natthawan Likitmongkonsakun, Tawan Limpiyakorn

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17α-ethynylestradiol (EE2) is a synthetic estrogen used as a key ingredient in an oral contraceptives pill. EE2 is an endocrine disrupting compound, high in estrogenic potency. Although EE2 exhibits low degree of biodegradability with common microorganisms in wastewater treatment plants (WWTPs), this compound can be biotransformed by ammonia-oxidizing bacteria (AOB) via a co-metabolism mechanism in WWTPs. This study aimed to investigate the effect of real wastewater on biotransformation of EE2 by AOB. A preliminary experiment on the effect of nitrite and pH levels on abiotic transformation of EE2 suggested that the abiotic transformation occurred at only pH <6.8. Biotransformation of EE2 under the presence of municipal or industrial wastewater demonstrated that different types of wastewater affect EE2 biotransformation differently. Organic matters in wastewater were believed to deteriorate EE2 biotransformation via the competition effect. At a lower initial ammonium concentration, EE2 biotransformation can be retarded and the extent of the deterioration was COD-concentration dependent. However, when an initial ammonium concentration was elevated, thisphenomena disappeared. This is because when increasing the amount of the primary substrate, more AMO enzymes can be produced resulting in unlimited transformation of all compounds in the tests reducing the competitive effect of organic matters on EE2.

Keywords: 17α-ethynylestradiol, ammonia-oxidizing bacteria, nitrifying activated sludge, wastewater.

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Authors: Rosarin Rujananon, Poonsuk Prasertsan, Amornrat Phongdara, Tanate Panrat, Jibin Sun, Sugima Rappert, An-Ping Zeng

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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.

Keywords: Recombinant E.coli, 1, 3-propanediol, glycerol, fusion protein.

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Authors: Ashish Kumar Nayak, Dhamodharan K., Ajay S. Kalamdhad

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The present study is aimed at alteration of sewage sludge into stable compost product using vermicomposting of sewage sludge mixed with cattle manure and saw dust in five different proportions based on C/N ratios (C/N 15 (R1), 20 (R2), 25 (R3) and 30 (R4); and control (R5)) by employing an epigeic earthworm Eisenia fetida. Higher reductions in C/N ratio, CO₂ evolution and OUR were observed in R4 demonstrated the compost stability. In addition, R4 proved to be best combination for the growth of the earthworms. In order to observe the optimal degradation, kinetics for degradation of organic matter in vermicomposting were quantitatively evaluated. An approach model was developed by assuming that composting process is carried out in a homogeneous way and the kinetics for decomposition reaction is represented by a Monod-type equation. The results exhibit comparable variations in the kinetic constants K_m and K₃ under varying parameters during vermicomposting process. Results suggested that higher R² value in R4, enhanced suitability towards Lineweaver-Burke plot. R4 yields higher degradability coefficient (K) reveals that the occurrence of optimal nutrient balance, which not only enhanced the affinity of enzymes towards substrate but also improved its degradation process. Therefore, it can be proved that R4 provided to be the best feed combination for vermicomposting process as compared to other reactors.

Keywords: Vermicomposting, Eisenia fetida, Sewage sludge, C/N ratio, Stability, Enzyme kinetics concept.

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Authors: P. Thongmuang, Y. Sudjaroen

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The methanolic extracts from seeds of tamarind (Tamarindus indica) was prepared by Soxhlet apparatus extraction and evaluated for total phenolic content by Folin-Ciocalteu method. Then, methanolic extract was screened biological activities (In vitro) for anti-melanogenic activity by tyrosinase inhibition test, antiinflammation activity by cyclooxygenase 1 (COX-1) and cyclooxygenase 2 (COX-2) inhibition test, and cytotoxic screening test with Vero cells. The results showed that total phenolic content, which contained in extract, was contained 27.72 mg of gallic acid equivalent per g of dry weight. The ability to inhibit tyrosinase enzyme, which exerted by Tamarind seed extracts (1 mg/ml) was 52.13 ± 0.42 %. The extract was not possessed inhibitory effect to COX-1 and COX-2 enzymes and cytotoxic effect to Vero cells. The finding is concludes that tested seed extract was possessed antimelanogenic activity with non-toxic effects. However, there was not exhibited anti-inflammatory activity. Further studies include the use of advance biological models to confirm this biological activity, as well as, the isolation and characterization of the purified compounds that it was contained.

Keywords: Tamarindus indica, anti-melanogenic, antiinflammatotion, cytotoxicity, seed, phenolic compounds.

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Authors: Surasak Siripornadulsil, Nutt Poomai, Wilailak Siripornadulsil

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Due to a high ethanol demand, the approach for  effective ethanol production is important and has been developed  rapidly worldwide. Several agricultural wastes are highly  abundant in celluloses and the effective cellulase enzymes do exist  widely among microorganisms. Accordingly, the cellulose  degradation using microbial cellulase to produce a low-cost substrate  for ethanol production has attracted more attention. In this  study, the cellulase producing bacterial strain has been isolated  from rich straw and identified by 16S rDNA sequence analysis as Acinetobacter sp. KKU44. This strain is able to grow and exhibit the cellulase activity. The optimal temperature for its growth and  cellulase production is 37°C. The optimal temperature of bacterial  cellulase activity is 60°C. The cellulase enzyme from  Acinetobacter sp. KKU44 is heat-tolerant enzyme. The bacterial culture of 36h. showed highest cellulase activity at 120U/mL when  grown in LB medium containing 2% (w/v). The capability of  Acinetobacter sp. KKU44 to grow in cellulosic agricultural wastes as a sole carbon source and exhibiting the high cellulase activity at high temperature suggested that this strain could be potentially developed further as a cellulose degrading strain for a production of low-cost substrate used in ethanol production. 

 

Keywords: Acinetobacter sp. KKU44, bagasse, cellulase enzyme, rice husk.

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Authors: T. N. M. El Sebai, A. A.Khattab, Wafaa M. Abd-El Rahim, H. Moawad

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Fungal mutant strains have produced cellulase and xylanase enzymes, and have induced high hydrolysis with enhanced of rice straw. The mutants were obtained by exposing Penicillium strain to UV-light treatments. Screening and selection after treatment with UV-light were carried out using cellulolytic and xylanolytic clear zones method to select the hypercellulolytic and hyperxylanolytic mutants. These mutants were evaluated for their cellulase and xylanase enzyme production as well as their abilities for biodegradation of rice straw. The mutant 12 UV/1 produced 306.21% and 209.91% cellulase and xylanase, respectively, as compared with the original wild type strain. This mutant showed high capacity of rice straw degradation. The effectiveness of tested mutant strain and that of wild strain was compared in relation to enhancing the composting process of rice straw and animal manures mixture. The results obtained showed that the compost product of inoculated mixture with mutant strain (12 UV/1) was the best compared to the wild strain and un-inoculated mixture. Analysis of the composted materials showed that the characteristics of the produced compost were close to those of the high quality standard compost. The results obtained in the present work suggest that the combination between rice straw and animal manure could be used for enhancing the composting process of rice straw and particularly when applied with fungal decomposer accelerating the composting process.

Keywords: Rice straw, composting, UV mutants, Penicillium.

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Authors: Tatsaporn Todhanakasem, Kamonchanok Areerat, Pornthap Thanonkeo, Roungdao KlinjapoandGlenn M. Young

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Rice bran has been abandoned as agricultural waste for million tonnes per year in Thailand, therefore they have been proposed to be utilized as a rich carbon source in the production of bioethanol. Many toxic compounds are possibly released during the pretreatment of rice bran prior the fermentation process. This study aims to analyze on the availability of toxic compounds and the amount of glucose obtained from 2 different pretreatments using sulfuric acid and mixed cellulase enzymes (without and with delignification/ activated charcoal). The concentration of furfural, 5- hydroxymethyl furfural (5-HMF), levulinic acid, vanillin, syringaldehyde and4-hydroxybenzaldehyde (4-HB) and the percent acetic acid were found to be 0.0517 ± 0.049 mg/L, 0.032 ± 0.06 mg/L, 21074 ± 1685.62 mg/L, 126.265 ± 6.005 mg/L, 2.89 ± 0.30 mg/L, 0.37 ± 0.031mg/L and 0.72% under the pretreatment process without delignification/ activated charcoal treatment and 384.47 ± 99.02 g/L, 0.068 mg/L, 142107.62 ± 8664.6 mg/L, 0.19 mg/L, 5.43 ± 3.29 mg/L, 4.80 ± 0.76 mg/L and 0.254% under the pretreatment process with delignification/ activated charcoal treatment respectively. The presence of high concentration of acetic acid was found to impede the growth of Zymomonas mobilis strain TISTR 551 despite the present of high concentration of levulinic acid. Z. mobilis strain TISTR 551 was found to produce 8.96 ± 4.06 g/L of ethanol under 4 days fementation period in biofilm stage in which represented 40% theoretical yield.

Keywords: Rice bran, Zymomonas mobilis, biofilm, ethanol.

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Authors: Fereshteh Shacheraghi, Mohammad Reza Shakibaie, Hanieh Noveiri

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Fourty one strains of ESBL producing P.aeruginosa which were previously isolated from burn patients in Kerman University general hospital, Iran were subjected to PCR, RFLP and sequencing in order to determine the type of extended spectrum β- lactamases (ESBL), the restriction digestion pattern and possibility of mutation among detected genes. DNA extraction was carried out by phenol chloroform method. PCR for detection of bla genes was performed using specific primer for each gene. Restriction Fragment Length Polymorphism (RFLP) for ESBL genes was carried out using EcoRI, NheI, PVUII, EcoRV, DdeI, and PstI restriction enzymes. The PCR products were subjected to direct sequencing of both the strands for identification of the ESBL genes.The blaCTX-M, blaVEB-1, blaPER-1, blaGES-1, blaOXA-1, blaOXA-4 and blaOXA-10 genes were detected in the (n=1) 2.43%, (n=41)100%, (n=28) 68.3%, (n=10) 24.4%, (n=29) 70.7%, (n=7)17.1% and (n=38) 92.7% of the ESBL producing isolates respectively. The RFLP analysis showed that each ESBL gene has identical pattern of digestion among the isolated strains. Sequencing of the ESBL genes confirmed the genuinety of PCR products and revealed no mutation in the restriction sites of the above genes. From results of the present investigation it can be concluded that blaVEB-1 and blaCTX-M were the most and the least frequently isolated ESBL genes among the P.aeruginosa strains isolated from burn patients. The RFLP and sequencing analysis revealed that same clone of the bla genes were indeed existed among the antibiotic resistant strains.

Keywords: ESBL genes, PCR, RFLP, Sequencing, P.aeruginosa

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Authors: Lebari B. Gboeloh

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Ficus exasperata is a plant used in the traditional management of malaria in south-south Nigeria. An investigation into the effects of the ethanolic extract of the leaf of the plant on some biochemical indices in albino mice infected with Plasmodium berghei (NK 65) was conducted. 48 mice with weight range of 13-23 g were grouped into six (A, B, C, D, E, and F). Each group contained 8 mice. Groups A, B, C, D and E were infected with blood containing the parasite. Group F was not infected and served as the normal control. On the 6^th day after infection, 4 mice from each group were sacrificed and blood samples are collected for investigation. The remaining mice in each group were treated. Mice in Groups A, B and C were administered orally with 200, 300 and 500 mg/kg body weight of Ficus exasperata respectively for six days. Group D was not treated while Group F was given distilled water. Group E was treated with 5 mg/kg body weight of chloroquine. On the 6^th day post treatment, these mice were sacrificed and blood samples were collected for biochemical analysis. The results indicated that on the 6^th day post inoculation, the levels of aspartate aminotransferase (AST), alkaline phosphatase (ALP) and alanine aminotransferase (ALT) in all the mice infected with the parasite were significantly (p < 0.05) elevated. However, on the 6^th day post administration of extract, the increased levels of AST, ALP and ALT were significantly (p < 0.05) reduced in groups administered with 300 and 500 mg/kg body weight of the extract compared with groups D and F. The reduction in the levels of these enzymes is an indication that F. exasperata have no hepatotoxic effect on the mice at the dose levels administered.

Keywords: Ficus exasperata, albino mice, Plasmodium berghei, biochemical parameters.

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Authors: Mohd Sidek Ahmad, Zainon Mohd Noor, Zaidah Zainal Ariffin

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Fibrin degradation is an important part in prevention or treatment of intravascular thrombosis and cardiovascular diseases. Plasmin like fibrinolytic enzymes has given new hope to patient with cardiovascular diseases by treating fibrin aggregation related diseases with traditional plasminogen activator which have many side effects. Various researches involving wide range of sources for production of fibrinolytic proteases, from bacteria, fungi, insects and fermented foods. But few have looked into endophytic fungi as a potential source. Sixteen (16) endophytic fungi were isolated from Hibiscus sp. leaves from six different locations in Shah Alam, Selangor. Only two endophytic fungi, FH3 and S13 showed positive fibrinolytic protease activities. FH3 produced 5.78cm and S13 produced 4.48cm on Skim Milk Agar after 4 days of incubation at 27°C. Fibrinolytic activity was observed; 3.87cm and 1.82cm diameter clear zone on fibrin plate of FH3 and S13 respectively. 18srRNA was done for identification of the isolated fungi with positive fibrinolytic protease. S13 had the highest similarity (100%) to that of Penicillium citrinum strain TG2 and FH3 had the highest similarity (99%) to that of Fusarium sp. FW2PhC1, Fusarium sp. 13002, Fusarium sp. 08006, Fusarium equiseti strain Salicorn 8 and Fungal sp. FCASAn-2. Media composition variation showed the effects of carbon nitrogen on protein concentration, where the decrement of 50% of media composition caused drastic decrease in protease of FH3 from 1.081 to 0.056 and also S13 from 2.946 to 0.198.

Keywords: Isolation, identification, fibrinolytic protease, endophytic fungi, Hibiscus leaves.

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Authors: Wehash, F. E., Ismail I. Abo-Ghanema, Rasha Mohamed Saleh

Abstract:

This study was conducted to evaluate the anti-diabetic properties of ethanolic extract of two plants commonly used in folk medicine, Mormodica charantia (bitter melon) and Trigonella foenum-graecum (fenugreek). The study was performed on STZinduced diabetic rats (DM type-I). Plant extracts of these two plants were given to STZ diabetic rats at the concentration of 500 mg/kg body weight ,50 mg/kg body weight respectively. Cidophage® (metformin HCl) were administered to another group to support the results at a dose of 500 mg/kg body weight, the ethanolic extracts and Cidophage administered orally once a day for four weeks using a stomach tube and; serum samples were obtained for biochemical analysis. The extracts caused significant decreases in glucose levels compared with diabetic control rats. Insulin secretions were increased after 4 weeks of treatment with Cidophage® compared with the control non-diabetic rats. Levels of AST and ALT liver enzymes were normalized by all treatments. Decreases in liver cholesterol, triglycerides, and LDL in diabetic rats were observed with all treatments. HDL levels were increased by the treatments in the following order: bitter melon, Cidophage®, and fenugreek. Creatinine levels were reduced by all treatments. Serum nitric oxide and malonaldehyde levels were reduced by all extracts. GSH levels were increased by all extracts. Extravasation as measured by the Evans Blue test increased significantly in STZ-induced diabetic animals. This effect was reversed by ethanolic extracts of bitter melon or fenugreek.

Keywords: Cidophage®, Diabetic rats, Mormodica charantia, Trigonella foenum-graecum

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Authors: Yemisi Rufina Alli Smith, Isaac Gbadura Adanlawo

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Liver disorders are one of the major problems of the world. Despite its frequent occurrence, high morbidity and high mortality, its medical management is currently inadequate. This study was designed to evaluate the hepatoprotective effect of saponin extract of the root of Garcinia kola on the integrity of the liver of paracetamol induced wistar albino rats. Twenty five (25) male adult wistar albino rats were divided into five (5) groups. Group I was the Control group that received distilled water only, group II was the negative control that received 2 g/kg of paracetamol on the 13th day, and group III, IV and V were pre-treated with 100, 200 and 400mg/kg of the saponin extract before inducing the liver damage on the 13th day with 2 g/kg of paracetamol. Twenty four (24) h after administration, the rats were sacrificed and blood samples were collected. The serum Alanine Transaminase (ALT), Aspartate Transaminase (AST), Alkaline Phosphatase (ALP) activities, Bilirubin and conjugated bilirubin, glucose and protein concentrations were evaluated. The liver was fixed immediately in Formalin and was processed and stained in Haematoxylin and Eosin (H&E). Administration of saponin extract from the root of Garcinia kola significantly decreased paracetamol induced elevated enzymes in the test group. Also histological observations showed that saponin extract of the root of Garcinia kola exhibited a significant liver protection against the toxicant as evident by the cells trying to return to normal. Saponin extract from the root of Garcinia kola indicated a protection of structural integrity of the hepatocytic cell membrane and regeneration of the damaged liver.

Keywords: Garcinia kola, Hepatoprotective, paracetamol, Saponin.

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Authors: Raad Nari, Maura Moriaty, Maha T. Barakat

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Introduction: Sodium-glucose co-transporter-2 (SGLT2) inhibitors are a new class of oral anti-diabetic drugs with a unique mechanism of action. They are used to improve glycaemic control in adults with type 2 diabetes by enhancing urinary glucose excretion. In the UAE, there has been certainly an increased use of these medications. As with any new medication, there are safety considerations related to their use in patients with type two diabetes. A retrospective study was conducted at the three main centres of the Imperial College London Diabetes Centre. Methodology: All patients in electronic database (Diamond) from October 2014 to October 2017 were included with a minimum of six months usage of sodium glucose co-transporter inhibitors that comprise canagliflozin, dapagliflozin and empagliflozin. There were 15 paired sample biochemical and clinical correlations. The analysis was done at the start of the study, three months and six months apart. SPSS version 24 was used for this study. Conclusion: This study of sodium glucose co-transporter-2 inhibitors used showed significant reductions in weight, glycated haemoglobin A1C, systolic and diastolic blood pressures. As the case with systematic reviews, there were similar changes in liver enzymes, raised total cholesterol, low density lipopoptein and high density lipoprotein. There was slight improvement in estimated glomerular filtration rate too. Our analysis also showed that they increased in the incidence of urinary tract symptoms and incidence of urinary tract infections.

Keywords: SGLT2 inhibitors dapagliflozin empagliflozin canagliflozin, adverse effects, amputation diabetic ketoacidosis DKA, urinary tract infection.

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Authors: N. V. Enwuru, B. Nkeki, E. A. Adekoya, O. A. Adebesin, B. O. Ojo, R. F. Peters, V. A. Aikhomu, U. E. Mendie, O. Akinloye

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Awareness about probiotic health benefits is increasing tremendously. However, cell viability is often low due to harsh conditions exposed during processing, handling, storage, and gastrointestinal transit. Thus, encapsulation is a promising technique that increases cell viability. The study aims to encapsulate the probiotic, evaluate its viability and cytotoxic activity of its postbiotic on the Michigan Cancer Foundation (MCF)-7 breast cancer cell line. Human and animal raw milk was sampled for lactic acid bacteria. Isolated bacteria were identified using conventional and VITEK 2 systems. The identified bacteria were encapsulated using the spray-drying method. The free and encapsulated probiotic cells were exposed to simulated gastric intestinal (SGI) fluid conditions and different storage conditions for their viability. The properties of the formed probiotic granules, their disintegration time, and the weight uniformity of the microcapsules were tested. Furthermore, the postbiotic of the free cells was extracted, and its cytotoxic effect on the MCF-7 breast cancer cell line was tested through [3-(4,5-dimethylthiazolyl-2)-2,5 diphenyltetrazolium bromide] (MTT) assay. The bacteria isolated were identified as Lactobacillus plantarum. The size of the formed probiotic granules ranges within 0.71-1.00 mm in diameter, and disintegration time ranges from 2.14 ± 0.045 to 2.91 ± 0.293 minutes, while the average weight is 502.1 mg. The viability of encapsulated cells stored at refrigerated condition (4oC) was higher than that of cells stored at room temperature (25 oC). The encapsulated probiotic cells exhibited better viability after exposure to SGI solution at different pH levels than free cells. The Postbiotic Metabolites (PM) of L. plantarum produced a cytotoxic effect that shows significant activity similar to 5FU, a standard antineoplastic agent. The inhibition concentration of 50% growth (IC50) of postbiotic metabolite was consistent with the IC50 of the positive control (Cisplatin). Lactobacillus plantarum postbiotic exhibited a cytotoxic effect on the MCF-7 breast cancer cell line and could be used as combined adjuvant therapy in breast cancer management. The microencapsulation technique protects the probiotics and maintains their viability.

Keywords: Cytotoxicity effect, encapsulation, postbiotic, probiotic.

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Authors: Ismail I. Abo Ghanema, Kadry M. Sadek

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Oxidative stress and overwhelming free radicals associated with diabetes mellitus are likely to be linked with development of certain complication such as retinopathy, nephropathy and neuropathy. Treatment of diabetic subjects with antioxidant may be of advantage in attenuating these complications. Olive leaf (Oleaeuropaea), has been endowed with many beneficial and health promoting properties mostly linked to its antioxidant activity. This study aimed to evaluate the significance of supplementation of Olive leaves extract (OLE) in reducing oxidative stress, hyperglycemia and hyperlipidemia in Sterptozotocin (STZ)- induced diabetic rats. After induction of diabetes, a significant rise in plasma glucose, lipid profiles except High density lipoproteincholestrol (HDLc), malondialdehyde (MDA) and significant decrease of plasma insulin, HDLc and Plasma reduced glutathione GSH as well as alteration in enzymatic antioxidants was observed in all diabetic animals. During treatment of diabetic rats with 0.5g/kg body weight of Olive leaves extract (OLE) the levels of plasma (MDA) ,(GSH), insulin, lipid profiles along with blood glucose and erythrocyte enzymatic antioxidant enzymes were significantly restored to establish values that were not different from normal control rats. Untreated diabetic rats on the other hand demonstrated persistent alterations in the oxidative stress marker (MDA), blood glucose, insulin, lipid profiles and the antioxidant parameters. These results demonstrate that OLE may be of advantage in inhibiting hyperglycemia, hyperlipidemia and oxidative stress induced by diabetes and suggest that administration of OLE may be helpful in the prevention or at least reduced of diabetic complications associated with oxidative stress.

Keywords: Diabetes mellitus, olive leaves, oxidative stress, red blood cells

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Authors: Abou-Baker Salim, A. A. K. Abou-Arab, Sherif R. Mohamed, T. A. Eldesouky

Abstract:

Pomegranate (Punica granatum L.) is an ancient fruit of great medical interest and rich source of antioxidants. Pesticides as dimethoate play a crucial role in the occurrence many diseases in plants, animal and human. Therefore the ability of Pomegranate (Punica granatum L.) to alleviate hepatotoxicity induced by organophosphate pesticide dimethoate was investigated. Albino male rats were divided randomly into 4 groups and kept at 7 animals per group in an environmentally controlled condition for 6 weeks. The first group was served as a control group (basal diet), the second group fed on basal diet supplemented with 5% freeze dried pomegranate seeds, the third group fed on 20 ppm dimethoate contaminated diet and the last group fed on dimethoate contaminated diet supplemented with 5% freeze dried pomegranate seeds. The results revealed that administration of dimethoate caused high significant increased in liver functions: alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) activities as well as lipid peroxide (malonaldhyde, MDA); on the other hand high significant decreased on glutathione (GSH), glutathione peroxidase (GPx), albumin and total protein were observed. However addition of 5% freeze dried pomegranate seeds significantly improved all previously mentioned parameters. These results indicate the dimethoate induced hepatotoxicity and highlight the protective effect of pomegranate seeds as a potential protective agent against dimethoate induced hepatotoxicity. This may be attributed to the powerful antioxidants (polyphenols, total phenols, and total flavonoids) which present in high levels in pomegranate as well as improving the immunity by activation of antioxidant enzymes GSH and GPx.

Keywords: Pomegranate, organophosphate pesticides, dimethoate, liver toxicity, free radicals, antioxidants.

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Authors: Gnanaprakash K, Madhusudhana Chetty C, Ramkanth S, Alagusundaram M, Tiruvengadarajan VS, Angala Parameswari S, Mohamed Saleem TS

Abstract:

Carbon tetrachloride (CCl4) is a well-known hepatotoxin and exposure to this chemical is known to induce oxidative stress and causes liver injury by the formation of free radicals. Flacourtia indica commonly known as 'Baichi' has been reported as an effective remedy for the treatment of a variety of diseases. The objective of this study was to investigate the hepatoprotective activity of aqueous extract of leaves of Flacourtia indica against CCl4 induced hepatotoxicity. Animals were pretreated with the aqueous extract of Flacourtia indica (250 & 500 mg/kg body weight) for one week and then challenged with CCl4 (1.5 ml/kg bw) in olive oil (1:1, v/v) on 7th day. Serum marker enzymes (ALP, AST, ALT, Total Protein & Total Bilirubin) and TBARS level (Marker for oxidative stress) were estimated in all the study groups. Alteration in the levels of biochemical markers of hepatic damage like AST, ALT, ALP, Total Protein, Total Bilirubin and lipid peroxides (TBARS) were tested in both CCl4 treated and extract treated groups. CCl4 has enhanced the AST, ALT, ALP and the Lipid peroxides (TBARS) in liver. Treatment of aqueous extract of Flacourtia indica leaves (250 & 500 mg/kg) exhibited a significant protective effect by altering the serum levels of AST, ALT, ALP, Total Protein, Total Bilirubin and liver TBARS. These biochemical observations were supported by histopathological study of liver sections. From this preliminary study it has been concluded that the aqueous extract of the leaves of Flacourtia indica protects liver against oxidative damages and could be used as an effective protector against CCl4 induced hepatic damage. Our findings suggested that Flacourtia indica possessed good hepatoprotective activity

Keywords: Carbon Tetrachloride, Flacourtia indica, Hepatoprotective activity, Oxidative stress

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