International Science Index

International Journal of Biotechnology and Bioengineering

Surface Modified Polyamidoamine Dendrimer with Gallic Acid Overcomes Drug Resistance in Colon Cancer Cells HCT-116
Cancer cells can develop resistance to conventional therapies especially chemotherapeutic drugs. Resistance to chemotherapy is another challenge in cancer therapeutics. Therefore, it is important to address this issue. Gallic acid (GA) is a natural plant compound that exhibits various biological properties including anti-proliferative, anti-inflammatory, anti-oxidant and anti-bacterial. Despite of the wide spectrum biological properties GA has cytotoxic response and low bioavailability. To overcome this problem, GA was conjugated with the Polyamidoamine(PAMAM) dendrimer for improving the bioavailability and efficient delivery in drug-resistant HCT-116 Colon Cancer cells. Gallic acid was covalently linked to 4.0 G PAMAM dendrimer. PAMAM dendrimer is well established nanocarrier but has cytotoxicity due to presence of amphiphilic nature of amino group. In our study we have modified surface of PAMAM dendrimer with Gallic acid and examine their anti-proliferative effects in drug-resistant HCT-116 cells. Further, drug-resistant colon cancer cells were established and thereafter treated with different concentration of PAMAM-GA to examine their anti-proliferative potential. Our results show that PAMAM-GA conjugate induces apoptotic cell death in HCT-116 and drug-resistant cells observed by Annexin-PI staining. In addition, it also shows that multidrug-resistant drug transporter P-gp protein expression was downregulated with increasing the concentration of GA conjugate. After that we also observed the significant difference in Rh123 efflux and accumulation in drug sensitive and drug-resistant cancer cells. Thus, our study suggests that conjugation of anti-cancer agents with PAMAM could improve drug resistant property and cytotoxic response to treatment of cancer.
Microbial Consortia with Biodegradation and Metals Bioleaching Potential from Estonian Graptolite Argillite
Estonian graptolite-argillite (GA) is enriched with several heavy metals (U, Mo, V, Pb, Co, Zn, Re, Ni, etc.) as sulfide minerals and/or in the composition of organometallic compounds. We have observed that heterotrophic facultative anaerobes and methanogenic archaea of argillite indigenous microbial community ARGCON5 are able to decompose the organic matter of GA under anaerobic conditions and at pH 7 resulting in methane and CO₂ generation with releasing metals, like Co, Ni, Mo, Zn. Some of the microorganisms present in the microbial community ARGCON5 can tolerate extremely high temperature. For example, with cloning and DNA sequencing of the mcrA gene, the presence of Methanosarcina thermophila TM-1 (GB: CP009501.1) was verified in GA samples sterilized at t=100 °C. By DNA sequence analysis of the 16S rRNA gene, Aneurinibacillus migulanus (GB: GU397386.1) and Bacillus fumarioli TN18 (GB: JQ415990.1) were identified in GA samples sterilized at t=121 °C. As anaerobic biodegradation occurs slowly in comparison with aerobic growth, majority of bioleaching experiments worldwide have been performed in aerobic environment. Therefore aerobic cultivation experiments were also performed for comparing the degradation efficiency of organic matter of GA and bioleaching of metals under anaerobic and aerobic conditions. In adaptive laboratory evolution experiments both the growth of the currently existing consortium ARGCON5 and new consortia enriched from different GA samples from drill-core (near Keila) and outcrops (from Pakri peninsula) were monitored. Aerobic cultivation experiments were performed via serial passages in liquid growth medium containing GA as the sole carbon source. Compared to the anaerobic cultivation, aerobic neutrophilic consortia obtained in adaptive laboratory evolution experiments by enrichment from various GA samples were more efficient in leaching As, Mo, Se, Sb, V. Taking together, our results indicate that both anaerobic and aerobic cultivation conditions can be used for decomposing of organic component of GA and bioleaching of metals from GA. We suggest that in order to obtain higher yields of bioleached metals anaerobic and aerobic cultivation conditions could be combined.
Heterotrophic Bioleaching of Secondary Sources Using Ammonia
With the number of easily accessible ores depleting, particularly in Europe, alternate sources are required to meet the increasing demand of critical metals. Secondary sources, such as mining and metallurgical residues, sludge effluents and shredded materials, whilst bearing lower metal loadings, are thus becoming of increasing economic interest. Traditional metallurgical techniques often require the use of strong acids or solvents to leach metals from sources, but are expensive, non-selective and often create more waste. Bioleaching offers an alternate, greener and more sustainable approach of metal extraction. Chemolithotrophic and heterotrophic microorganisms can leach metals from materials through direct microbe interactions or the production of a biolixiviant solution to extract and chelate metals from the matrix. This research investigates the use of biogenic ammonia on six secondary materials, comprised of metallurgical residues, industrial sludges and automotive shredder residues, using an ureolytic strain of Bacillus. We also aimed to improve leaching by varying media components (yeast extract/urea), pulp density and impact of cells with and without lixiviant. We observed moderate to high yields (10-40%) and very high selectivity (>97% against non target metals) of copper, nickel and zinc leached from the metallurgical residues and sludges. For the automotive shredder residue, we observed higher degrees of leaching (>70 %) and selectivity (>99 %) and interestingly, this was higher in the biological system compared to the chemical counterpart. Furthermore, pre treatment using ammonia bioleaching, combined with subsequent next generation solvo/hydro metallurgical techniques enhanced leaching in some of the materials tested. Finally, mineralogical assessments (SEM and XRD) were also undertaken to determine whether the residual material was suitable for reuse in building materials. These results provide important findings for future sustainable metal recovery from secondary materials.
Accurate Ratio Computation Using Abstract Chemical Reaction Networks
Ratio computation is an important computation on the molecular level and it has been conjectured that biological systems have evolved motifs to execute ratio computation. We present new results on how abstract chemical reaction networks (ACRNs), viz., catalysis, annihilation and degradation, can be used to implement circuits that accurately compute ratio of two input signals. The input signals can be either constant values scalars or time-varying scalars or polynomials. We also characterise the robustness of our circuits to parameter variations, as would be encountered in wet-lab implementations. Our ACRNs can be implemented in wet-lab via DNA strand displacement techniques or genelets or DNA Toolbox.
Improved Predictive Models for the in vivo Reverse-Engineering and Modeling Assessment Network Using Nonlinear Optimisation
Cellular complexity stems from the interactions among thousands of different molecular species. Thanks to the emerging fields of systems and synthetic biology, scientists are beginning to unravel these regulatory, signaling, and metabolic interactions and to understand their coordinated action. Reverse engineering of biological networks has several benefits but a poor quality of data combined with the difficulty in reproducing it limits the applicability of these methods. A few years back, many of the commonly used predictive algorithms were tested on a network constructed in the yeast Saccharomyces cerevisiae (S. cerevisiae) to resolve this issue. The network was a synthetic network of five genes regulating each other for the so-called in vivo reverse-engineering and modeling assessment (IRMA). The network was constructed in S. cereviase since it is a simple and well characterized organism. The synthetic network included a variety of regulatory interactions, thus capturing the behaviour of larger eukaryotic gene networks on a smaller scale. We derive a new set of algorithms by solving a nonlinear optimization problem and show how these algorithms outperform other algorithms on these datasets.
Design and Fabrication of Stiffness Reduced Metallic Locking Compression Plates through Topology Optimization and Additive Manufacturing
Bone fixation implants currently used to treat traumatic fractured bones and to promote fracture healing are built with biocompatible metallic materials such as stainless steel, cobalt chromium and titanium and its alloys (e.g., CoCrMo and Ti6Al4V). The noticeable stiffness mismatch between current metallic implants and host bone associates with negative outcomes such as stress shielding which causes bone loss and implant loosening leading to deficient fracture treatment. This paper, part of a major research program to design the next generation of bone fixation implants, describes the combined use of three-dimensional (3D) topology optimization (TO) and additive manufacturing powder bed technology (Electron Beam Melting) to redesign and fabricate the plates based on the current standard one (i.e., locking compression plate). Topology optimization is applied with an objective function to maximize the stiffness and constraint by volume reductions (i.e., 25-75%) in order to obtain optimized implant designs with reduced stress shielding phenomenon, under different boundary conditions (i.e., tension, bending, torsion and combined loads). The stiffness of the original and optimised plates are assessed through a finite-element study. The TO results showed actual reduction in the stiffness for most of the plates due to the critical values of volume reduction. Additionally, the optimized plates fabricated using powder bed techniques proved that the integration between the TO and additive manufacturing presents the capability of producing stiff reduced plates with acceptable tolerances.
Bean in Turkey: Characterization, Inter Gene Pool Hybridization Events, Breeding, Utilizations
Turkey is considered a bridge between Europe, Asia, and Africa and possibly played an important role in the distribution of many crops including common bean. Hundreds of common bean landraces can be found in Turkey, particularly in farmers’ fields, and they consistently contribute to the overall production. To investigate the existing genetic diversity and hybridization events between the Andean and Mesoamerican gene pools in the Turkish common bean, 188 common bean accessions (182 landraces and 6 modern cultivars as controls) were collected from 19 different Turkish geographic regions. These accessions were characterized using phenotypic data (growth habit and seed weight), geographic provenance, 12557 high-quality whole-genome DArTseq markers, and 3767 novel DArTseq loci were also identified. The clustering algorithms resolved the Turkish common bean landrace germplasm into the two recognized gene pools, the Mesoamerican and Andean gene pools. Hybridization events were observed in both gene pools (14.36% of the accessions) but mostly in the Mesoamerican (7.97% of the accessions), and was low relative to previous European studies. The lower level of hybridization witnessed the existence of Turkish common bean germplasm in its original form as compared to Europe. Mesoamerican gene pool reflected a higher level of diversity, while the Andean gene pool was predominant (56.91% of the accessions), but genetically less diverse and phenotypically more pure, reflecting farmers greater preference for the Andean gene pool. We also found some genetically distinct landraces and overall, a meaningful level of genetic variability which can be used by the scientific community in breeding efforts to develop superior common bean strains.
In Vitro Axillary Shoot Regeneration of Black Chickpea (Cicer arietinum L.) Using Kinetin
Black chickpea (Cicer arietinum) is an important dietary legume in Indian sub-continent. The seeds are relatively small and rich in protein and used as food and medicinal in different forms. In this study, different explants of black chickpea were used for in vitro shoot regeneration on Murashige and Skoog (MS) medium supplemented with different concentrations of Kinetin (0.25, 0.50, 1.0, 1.50, 2.0, 3.0 mg/l). Shoot tip and cotyledonary node explants were taken form 5-7 days old grown seedlings and cultured on kinetin based MS medium for 8 weeks. Shoot regeneration from all explants started within one week with multiple shoot induction in two weeks. Maximum number of 7.16 shoot with longer shoot of 4.41 cm were recorded from seeds explant. Comparision of kinetin doses revealed the increase in number of shoots with increased kinetin concentration with maximum number of 6.10 shoot were obtained on medium containing 3.0 mg/l kinetin. Interactive effect of explant x kinetin revealed the maximum number of shoots and longer length on MS medium with 2.0 mg/l kinetin for all explants. In vitro regenerated shoots were rooted on MS medium supplemented with different concentrations of Indole-6-Butyric Acid (IBA) followed by successful adaptation of plantlets in the pots. This regeneration protocol can be used for the application of other biotechnological techniques like genetic transformation.
Plant Mediated RNAi Approach to Knock Down Ecdysone Receptor Gene of Colorado Potato Beetle
RNA interference (RNAi) has proved its usefulness in functional genomic research on insects recently and is considered potential strategy in crop improvement for the control of insect pests. The different insect pests incur significant losses to potato yield worldwide, Colorado Potato Beetle (CPB) being most notorious one. The present study focuses to knock down highly specific 20-hydroxyecdysone hormone-receptor complex interaction by using RNAi approach to silence Ecdysone receptor (EcR) gene of CPB in transgenic potato plants expressing dsRNA of EcR gene. The partial cDNA of Ecdysone receptor gene of CPB was amplified using specific primers in sense and anti-sense orientation and cloned in pRNAi-GG vector flanked by an intronic sequence (pdk). Leaf and internodal explants of Lady Olympia, Agria and Granola cultivars of potato were infected with Agrobacterium strain LBA4404 harboring plasmid pRNAi-CPB, pRNAi-GFP (used as control). Neomycin phosphotransferase (nptII) gene was used as a plant selectable marker at a concentration of 100 mg L⁻¹. The primary transformants obtained have shown proper integration of T-DNA in plant genome by standard molecular analysis like polymerase chain reaction (PCR), real-time PCR, Sothern blot. The transgenic plants developed out of these cultivars are being evaluated for their efficacy against larvae as well adults of CPB. The transgenic lines are expected to inhibit expression of EcR protein gene, hindering their molting process, hence leading to increased potato yield.
Solubility and Dissolution Enhancement of Poorly Soluble Drugs Using Biosericin
Currently, sericin is being treated as waste of sericulture industry, especially at reeling process. Looking at prospective physicochemical properties, an attempt has been made to explore pharmaceutical applications of sericin waste in fabrication of medicated solid dispersions. Solid dispersions (SDs) of poorly soluble drugs (Lornoxicam, Meloxicam & Felodipine) were prepared by spray drying, solvent evaporation, ball milling and physical kneading in mass ratio of drug: sericin (1:0.5, 1:1, 1:1.5, 1:2, 1:2.5 and 1:3 w/w) and were investigated by solubility, ATR-FTIR, XRD and DSC, micromeritics and tablettability, surface morphology and in-vitro dissolution. It has been observed that sericin improves solubility of drugs by 8 to 10 times compared to pure drugs. The presence of hydrogen bonding between drugs and sericin was confirmed from the ATR-FTIR spectra. Amongst these methods, spray dried (1:2 w/w) SDs showed fully amorphous state representing molecularly distributed drug as confirmed from XRD and DSC study. Spray dried meloxicam SDs showed better compressibility and compactibility. The microphotograph of spray dried batches of lornoxicam (SDLX) and meloxicam SDs (SDMX) showed bowl shaped, and bowl plus spherical particles respectively, while spray dried felodipine SDs (SDFL) showed spherical shape. The SDLX, SDMX and SDFL (1:2 w/w) displayed better dissolution performance than other methods. Conclusively, hydrophilic matrix of sericin can be used to deliver poor water soluble drugs and its aerodynamic shape may show a great potential for various drug deliveries. If established as pharmaceutical excipient, sericin holds a potential to revolutionise economics of pharmaceutical industry, and sericulture farming, especially of Asian countries.
Statistical Optimization and Production of Rhamnolipid by P. aeruginosa PAO1 Using Prickly Pear Peel as a Carbon Source
Production of rhamnolipids by Pseudomonas aeruginosa has attracted a growing interest during the last few decades due to its high productivity compared with other microorganisms. In the current work, rhamnolipids production by P. aeruginosa PAO1 was statistically modeled using Taguchi orthogonal array, numerically optimized and validated. Prickly Pear Peel (Opuntia ficus-indica) has been used as a carbon source for production of rhamnolipid. Finally, the optimum conditions for rhamnolipid production were applied in 5L working volume bioreactors at different aerations, agitation and controlled pH for maximum rhamnolipid production. In addition, kinetic studies of rhamnolipids production have been reported. At the end of the batch bioreactor optimization process, rhamnolipids production by P. aeruginosa PAO1 has reached the worldwide levels and can be applied for its industrial production.
Plantlet Regeneration from Zygotic Embryos of Securidaca longepedunculata Fresen
Securidaca longepedunculata Fresen (Violet tree) belongs to the family Polygalaceae characterised by papillionaceous purplish flowers. This medicinally valued plant disappears at an alarming rate due to intensified anthropopressure particularly the unregulated manner of subterranean plant parts' collection from natural stands. Some indiscriminately harvested plants bear seeds containing both mature and immature zygotic embryos that are often discarded. Here, such seeds are collected for this experiment. Seeds were collected, washed, de-coated, and dipped in 70 % (v/v) ethanol for 30 s followed by rising in 5 % solution sodium hypochlorite, containing two drops of tween 20, for another 25 min. Mature zygotic embryos (MZEs) were excised from seeds and cultured in two basal media (MS and B5), three carbon sources (sucrose, glucose and fructose) at five concentrations (0-40 g/L) while immature zygotic embryos (iMZEs) were composed on similar basal media and carbon source supplemented with 0-2 mg/L Benzylaminopurine (BAP) and 0-2 mg/L Indole acetic acid (IAA). MZEs cultured on MS + 30g/L sucrose differed significantly from other treatments at p≤0.05 with maximum percent sprouting (85.24± 5.67 %) and shoot length (7.53±0.67 cm). MZEs culture had the maximum percent sprouting (85.24± 5.67 %) and shoot length (7.53±0.67 cm) in medium containing MS+ 30g L-1 sucrose. iMZEs on the other hand had maximum growth on MS + 40g/L sucrose supplemented with 1.5 mg/L IAA+ 1.0 mg/L BAP. This study is a geared towards creating an alternative path for the maximum production of plants in vitro, thereby, preventing the plants from disappearing.
Smart Polymeric Nanoparticles Loaded with Vincristine Sulfate for Applications in Breast Cancer Drug Delivery in MDA-MB 231 and MCF7 Cell Lines
Stimuli-responsive nanomaterials play an essential role in loading, transporting and well-distribution of anti-cancer compounds in the cellular surroundings. The outstanding properties as the Lower Critical Solution Temperature (LCST), hydrolytic cleavage and protonation/deprotonation cycle, govern the release and delivery mechanisms of payloads. In this contribution, we experimentally determine the load efficiency and release of antineoplastic Vincristine Sulfate into PNIPAM-Interpenetrated-Chitosan (PIntC) nanoparticles. Structural analysis was performed by Fourier Transform Infrared Spectroscopy (FT-IR) and Proton Nuclear Magnetic Resonance (1HNMR). ζ-Potential (ζ) and Hydrodynamic diameter (DH) measurements were monitored by Electrophoretic Mobility (EM) and Dynamic Light scattering (DLS) respectively. Mathematical analysis of the release pharmacokinetics reveals a three-phase model above LCST, while a monophasic of Vincristine release model was observed at 32 °C. Cytotoxic essays reveal a noticeable enhancement of Vincristine effectiveness at low drug concentration on HeLa cervix cancer and MDA-MB-231 breast cancer.
LIZTOXD: Inclusive Lizard Toxin Database by Using MySQL Protocol
LIZTOXD provides a single source of high-quality information about proteinaceous lizard toxins that will be an invaluable resource for pharmacologists, neuroscientists, toxicologists, medicinal chemists, ion channel scientists, clinicians, and structural biologists. We will provide an intuitive, well-organized and user-friendly web interface that allows users to explore the detail information of Lizard and toxin proteins. It includes common name, scientific name, entry id, entry name, protein name and length of the protein sequence. The utility of this database is that it can provide a user-friendly interface for users to retrieve the information about Lizard, toxin and toxin protein of different Lizard species. These interfaces created in this database will satisfy the demands of the scientific community by providing in-depth knowledge about Lizard and its toxin. In the next phase of our project we will adopt methodology and by using A MySQL and Hypertext Preprocessor (PHP) which and for designing Smart Draw. A database is a wonderful piece of equipment for storing large quantities of data efficiently. The users can thus navigate from one section to another, depending on the field of interest of the user. This database contains a wealth of information on species, toxins, toxins, clinical data etc. LIZTOXD resource that provides comprehensive information about protein toxins from lizard toxins. The combination of specific classification schemes and a rich user interface allows researchers to easily locate and view information on the sequence, structure, and biological activity of these toxins. This manually curated database will be a valuable resource for both basic researchers as well as those interested in potential pharmaceutical and agricultural applications of lizard toxins.
Encryption and Decryption of Nucleic Acid Using Deoxyribonucleic Acid Algorithm
The deoxyribonucleic acid text provides a single source of high-quality Cryptography about Deoxyribonucleic acid sequence for structural biologists. We will provide an intuitive, well-organized and user-friendly web interface that allows users to encrypt and decrypt Deoxy Ribonucleic Acid sequence text. It includes complex, securing by using Algorithm to encrypt and decrypt Deoxy Ribonucleic Acid sequence. The utility of this Deoxy Ribonucleic Acid Sequence Text is that, it can provide a user-friendly interface for users to Encrypt and Decrypt store the information about Deoxy Ribonucleic Acid sequence. These interfaces created in this project will satisfy the demands of the scientific community by providing fully encrypt of Deoxy Ribonucleic Acid sequence during this website. We have adopted a methodology by using C# and Active Server Page.NET for programming which is smart and secure. Deoxy Ribonucleic Acid sequence text is a wonderful piece of equipment for encrypting large quantities of data, efficiently. The users can thus navigate from one encoding and store orange text, depending on the field for user’s interest. Algorithm classification allows a user to Protect the deoxy ribonucleic acid sequence from change, whether an alteration or error occurred during the Deoxy Ribonucleic Acid sequence data transfer. It will check the integrity of the Deoxy Ribonucleic Acid sequence data during the access.
Optimization of Hepatitis B Surface Antigen Purifications to Improving the Production of Hepatitis B Vaccines on Pichia pastoris
Hepatitis B is a liver inflammatory disease caused by hepatitis B virus (HBV). This infection can be prevented by vaccination which contains HBV surface protein (sHBsAg). However, vaccine supply is limited. Several attempts have been conducted to produce local sHBsAg. However, the purity degree and protein yield are still inadequate. Therefore optimization of HBsAg purification steps is required to obtain high yield with better purification fold. In this study, optimization of purification was done in 2 steps, precipitation using variation of NaCl concentration (0,3 M; 0,5 M; 0,7 M) and PEG (3%, 5%, 7%); ion exchange chromatography (IEC) using NaCl 300-500 mM elution buffer concentration.To determine HBsAg protein, bicinchoninic acid assay (BCA) and enzyme-linked immunosorbent assay (ELISA) was used in this study. Visualization of HBsAg protein was done by SDS-PAGE analysis. Based on quantitative analysis, optimal condition at precipitation step was given 0,3 M NaCl and PEG 3%, while in ion exchange chromatography step, the optimum condition when protein eluted with NaCl 500 mM. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis indicates that the presence of protein HBsAg with a molecular weight of 25 kDa (monomer) and 50 kDa (dimer). The optimum condition for purification of sHBsAg produced in Pichia pastoris gave a yield of 47% and purification fold 17x so that it would increase the production of hepatitis B vaccine to be more optimal.
Effect of Media Osmolarity on Vi Biosynthesis on Salmonella enterica serovar Typhi Strain C6524 Cultured on Batch System
Typhoid fever disease can be prevented by using a polysaccharide-based vaccine Vi which is a virulence factor of S.typhi. To produce high yield Vi polysaccharide from bacteria, it is important to know the biosynthesis of Vi polysaccharide and the regulators involved. In the In vivo condition, S. typhi faces different osmolarity, and the bacterial two-component system OmpR-EnvZ, regulate by up and down Capsular Vi polysaccharide biosynthesis. A high yielded Vi Polysaccharide strain, S. typhi strain C6524 used to study the effect of media osmolarity on Vi polysaccharide biosynthesis and the osmoregulation pattern of S. typhi strain C6524. The methods were performed by grown S. typhi strain C6524 grown on medium with 50 mM, 100 mM, and 150 mM osmolarity with the batch system. Vi polysaccharide concentration was measured by ELISA method. For further investigation of the osmoregulation pattern of strain C6524, the osmoregulator gene, OmpR, has been isolated and sequenced using the specific primer of the OmpR gene. Nucleotide sequence analysis is done with BLAST and Lallign. Amino Acid sequence analysis is done with Prosite and Multiple Sequence Alignment. The results of cultivation showed the average content of polysaccharide Vi for 50 mM, 100 mM, and 150 mM osmolarities 11.49 μg/mL, 12.06 μg/mL, and 14.53 μg/mL respectively. Analysis using Anova stated that the osmolarity treatment of 150 mM significantly affects Vi content. Analysis of nucleotide sequences shows 100% identity between S. typhi strain C6524 and Ty2. Analysis of amino acid sequences shows that the OmpR response regulator protein of the C6524 strain also has a α4-β5-α5 motif which is important for the regulatory activation system when phosphorylation occurs by domain kinase. This indicates that the regulator osmolarity response of S. typhi strain C6524 has no difference with the response regulator owned by S. typhi strain Ty2. A high Vi response rate in the 150 mM osmolarity treatment requires further research for RcsB-RcsC, another two-component system involved in Vi Biosynthesis.
Genetic Polymorphism and Insilico Study Epitope Block 2 MSP1 Gene of Plasmodium falciparum Isolate Endemic Jayapura
Malaria is an infectious disease caused by Plasmodium sp. This disease has a high prevalence in Indonesia, especially in Jayapura. The vaccine that is currently being developed has not been effective in overcoming malaria. This is due to the high polymorphism in the Plasmodium genome especially in areas that encode Plasmodium surface proteins. Merozoite Surface Protein 1 (MSP1) Plasmodium falciparum is a surface protein that plays a role in the invasion process in human erythrocytes through the interaction of Glycophorin A protein receptors and sialic acid in erythrocytes with Reticulocyte Binding Proteins (RBP) and Duffy Adhesion Protein (DAP) ligands in merozoites. MSP1 can be targeted to be a specific antigen and predicted epitope area which will be used for the development of diagnostic and malaria vaccine therapy. MSP1 consists of 17 blocks, each block is dimorphic, and has been marked as the K1 and MAD20 alleles. Exceptions only in block 2, because it has 3 alleles, among others K1, MAD20 and RO33. These polymorphisms cause allelic variations and implicate the severity of patients infected P. falciparum. In addition, polymorphism of MSP1 in Jayapura isolates has not been reported so it is interesting to be further identified and projected as a specific antigen. Therefore, in this study, we analyzed the allele polymorphism as well as detected the MSP1 epitope antigen candidate on block 2 P. falciparum. Clinical samples of selected malaria patients followed the consecutive sampling method, examining malaria parasites with blood preparations on glass objects observed through a microscope. Plasmodium DNA was isolated from the blood of malarial positive patients. The block 2 MSP1 gene was amplified using PCR method and cloned using the pGEM-T easy vector then transformed to TOP'10 E.coli. Positive colonies selection was performed with blue-white screening. The existence of target DNA was confirmed by PCR colonies and DNA sequencing methods. Furthermore, DNA sequence analysis was done through alignment and formation of a phylogenetic tree using MEGA 6 software and insilico analysis using IEDB software to predict epitope candidate for P. falciparum. A total of 15 patient samples have been isolated from Plasmodium DNA. PCR amplification results show the target gene size about ± 1049 bp. The results of MSP1 nucleotide alignment analysis reveal that block 2 MSP1 genes derived from the sample of malarial patients were distributed in four different allele family groups, K1 (7), MAD20 (1), RO33 (0) and MSP1_Jayapura (10) alleles. The most commonly appears of the detected allele is MSP1_Jayapura single allele. There was no significant association between sex variables, age, the density of parasitemia and alel variation (Mann Whitney, U > 0.05), while symptomatic signs have a significant difference as a trigger of detectable allele variation (U < 0.05). In this research, insilico study shows that there is a new epitope antigen candidate from the MSP1_Jayapura allele and it is predicted to be recognized by B cells with 17 amino acid lengths in the amino acid sequence 187 to 203.
rhCollagen-Based BioInk for 3D Bio‑Printing of Tissues and Organs
Collagen comprises the major component of connective tissues. Due to its dominance and unique characteristics, Type I collagen is frequently selected from a variety of biocompatible materials for use in tissue repair to provide structural integrity, induce cellular infiltration and promote tissue regeneration. Currently, collagen is primarily recycled from animal tissues. These sources suffer from several disadvantages, including consistency of the products, structural damage to the molecule, allergic response, risk of disease transmission and exposure to residual components, including growth factors originating from the source tissue. CollPlant’s unique technology enables to effectively produce, naïve human Type I collagen (rhCollagen) from tobacco plants. Two genes coding for the two human type I collagen chains, COL1 and COL2 were expressed, along with three human genes coding for the two modifying enzymes, proline hydroxylase (P4H) and lysine hydroxylase (LH3) which are critical to collagen maturation. The plant extracted rhCollagen forms thermally stable helical structures, preserving integrin and other essential binding sites. It fibrillates, and demonstrates bioactivity resembling that of native collagen. The protein is purified to homogeneity through a cost‑effective industrial process. The purified rhCollagen was evaluated in many models showing superiority over tissue extracted collagen in supporting viability and proliferation of cell types such as fibroblasts, endothelial cells, epithelial cells, and keratinocytes. An excellent safety profile of the material was demonstrated using standard assays, including advantage over bovine collagen in a T-cell immunogenicity profiling. Support for tissue regeneration was demonstrated in vivo showing superiority over tissue-derived collagen in both physical properties and biologic functionality. The unique biological and physical properties of rhCollagen were translated into approved medical devices which were already used to treat hundreds of humans in the fields of advanced wound care and orthobiologics. Recently, 3D bio‑printing is gaining momentum in many medicinal applications to address the need for complex scaffolds, tissues, and organs suitable for transplantation. Specifically for 3D application, the use of BioInk that is based on tissue extracted collagen is often limited due to its temperature-sensitive gelation kinetics at physiological conditions that require printing at low temperatures. For that purpose, rhCollagen was modified chemically to adopt the biological molecules for printing, such that the BioInk maintains controlled fluidity during printing, and cures to form hydrogel when irradiated by light ranging from UV to visible light. The unique viscosity and shear thinning properties of the modified rhCollagen allow the flexibility to easily formulate BioInks for different printing technologies including extrusion, ink-jet, Laser Induced Forward Transfer (LIFT) and Stereolithography. The control of chemical modification in combination with illumination energy allows tight control on the physical properties of the resulting scaffolds to match natural tissues properties, from stiff cartilage to soft adipose tissues. BioInks formulated from rhCollagen were evaluated with all major printing technologies and exhibited the required physical properties as well as excellent support for cells' proliferation and vitality including a series of primary and differentiated human cells. In conclusion, rhCollagen BioInk formulations provide a superior solution for 3D bioprinting of tissues and organs.
Impact of Autoclave Sterilization of Gelatin on Endotoxin Level and Physical Properties Compared to Surfactant Purified Gelatins
Introduction and Purpose: Endotoxins are found in the outer membrane of gram-negative bacteria and have profound in vitro and in vivo responses. They can trigger strong immune responses and negatively affect various cellar activities particular cells expressing toll-like receptors. They are therefore unwanted contaminants of biomaterials sourced from natural raw materials, and their activity must be as low as possible. Collagen and gelatin are natural extracellular matrix components and have, due to their low allergenic potential, suitable biological properties, and tunable physical characteristics, high potential in biomedical applications. The purpose of this study was to determine the influence of autoclave sterilization of gelatin on physical properties and endotoxin level compared to surfactant purified gelatin. Methods: Type A gelatin from Sigma-Aldrich (G1890) with endotoxin level of 35000 endotoxin units (EU) per gram gelatin and type A gelatins from Rousselot Gent with endotoxin activity of 30000 EU per gram were used. A 10 w/w% G1890 gelatin solution was autoclave sterilized during 30 minutes at 121°C and 1 bar over pressure. The physical properties and the endotoxin level of the sterilized G1890 gelatin were compared to a type A gelatin from Rousselot purified with Triton X100 surfactant. The Triton X100 was added to a concentration of 0.5 w/w% which is above the critical micellar concentration. The gelatin surfactant mixtures were kept for 30-45 minutes under constant stirring at 55-60°C. The Triton X100 was removed by active carbon filtration. The endotoxin levels of the gelatins were measured using the Endozyme recombinant factor C method from Hyglos GmbH (Germany). Results and Discussion: Autoclave sterilization significantly affect the physical properties of gelatin. Molecular weight of G1890 decreased from 140 to 50kDa, and gel strength decreased from 300 to 40g. The endotoxin level of the gelatin reduced after sterilization from 35000 EU/g to levels of 400-500 EU/g. These endotoxin levels are however still far above the upper endotoxin level of 0.05 EU/ml, which resembles 5 EU/g gelatin based on a 1% gelatin solution, to avoid cell proliferation alteration. Molecular weight and gel strength of Rousselot gelatin was not altered after Triton X100 purification and remained 150kDa and 300g respectively. The endotoxin levels of Triton X100 purified Rousselot gelatin was < 5EU/g gelatin. Conclusion: Autoclave sterilization of gelatin is, in comparison to Triton X100 purification, not efficient to inactivate endotoxin levels in gelatin to levels below the upper limit to avoid cell proliferation alteration. Autoclave sterilization gave a significant decrease in molecular weight and gel strength which makes autoclave sterilized gelatin, in comparison to Triton X100 purified gelatin, not suitable for 3D printing.
Drop-on-Demand Bioprinting Process for Single-Spheroid Deposition and Immobilization
Spheroids are widely used in manifold bioprinting applications, yet, to the author’s best knowledge, the spatially controlled deposition of separated spheroids at defined locations within a three-dimensional scaffold remains one major challenge. This current study examined an automated bioprinting process, based on a piezo-electric drop-on-demand dispenser, combined with an optical detection setup, allowing the contact-free deposition of small volume droplets carrying single-spheroids. The dispenser was used to produce liquid droplets with a typical volume of 10 nl, while its nozzle was continuously monitored with a magnifying lens setup. A software algorithm was used to process respective images of the dispenser nozzle, for the detection of single-spheroids present within the liquid volume of subsequent ejected droplets. A downstream vacuum aspirator allowed selection of ejected droplets, by discarding void droplets on the fly, while exclusively single-spheroid laden droplets were delivered to the substrate surface. The whole system was mounted on a three-axis robotic stage for coordinate controlled deposition of these droplets. In an application example, the process was used for the production of HUVEC spheroid arrays for comparative investigations and quantifications of angiogenesis processes in fibrin and collagen hydrogels. In sum, the process has proven potential for a flexible usage to place single-spheroid laden droplets in a contact-free manner onto prepared hydrogel surfaces, as well as immobilization for complete embedding in hydrogels of these structures including following incubation. So far, deposition accuracy, together with a resolution limit of 500 µm remained challenges for further improvements. Nevertheless, the controlled distribution of spheroids within hydrogels may allow systematic investigations of spheroid interactions, improve tissue engineering approaches, as well as bioprinting of organs-on-chip and large-scale tissue constructs with spheroids as distinct building blocks.
Process Development of pVAX1/lacZ Plasmid DNA Purification Using Design of Experiment
Third generation of vaccines is based on gene therapy where DNA is introduced into patients. The antigenic or therapeutic proteins encoded from transgenes DNA triggers an immune-response to counteract various diseases. Moreover, DNA vaccine offers the customization of its ability on protection and treatment with high stability. The production of DNA vaccines become of interest. According to USFDA guidance for industry, the recommended limits for impurities from host cell are lower than 1%, and the active conformation homogeneity supercoiled DNA, is more than 80%. Thus, the purification strategy using two-steps chromatography has been established and verified for its robustness. Herein, pVax1/lacZ, a pre-approved USFDA DNA vaccine backbone, was used and transformed into E. coli strain DH5α. Three purification process parameters including sample-loading flow rate, the salt concentration in washing and eluting buffer, were studied and the experiment was designed using response surface method with central composite face-centered (CCF) as a model. The designed range of selected parameters was 10% variation from the optimized set point as a safety factor. The purity in the percentage of supercoiled conformation obtained from each chromatography step, AIEX and HIC, were analyzed by HPLC. The response data were used to establish regression model and statistically analyzed followed by Monte Carlo simulation using SAS JMP. The results on the purity of the product obtained from AIEX and HIC are between 89.4 to 92.5% and 88.3 to 100.0%, respectively. Monte Carlo simulation showed that the pVAX1/lacZ purification process is robust with confidence intervals of 0.90 in range of 90.18-91.00% and 95.88-100.00%, for AIEX and HIC respectively.
Optimization of Tangential Flow Filtration Process for Purifying DNA Vaccine
Nowadays, DNA vaccines become an interesting subject in the third vaccine generation. The platform of DNA vaccines production has been developed and its downstream process becomes challenging due to the quality of the products in terms of purity and percentage of supercoiled DNA. To overcome these challenges, tangential flow filtration (TFF), which is involved in the purification process, could be used since it provides effective separation of impurity prior to performing further purification steps. However, operating conditions of TFF is varied based on several factors such as sizes of target particle and impurities, a concentration of solution as well as a concentration polarization on the membrane surface. In this study, pVAX1/lacZ was used as a model of TFF optimization in order to prevent a concentration polarization that can lead to the membrane fouling and also minimize a diafiltration volume while maintaining the maximum permeate flux resulting in proper operating times and buffer volume. By using trans membrane pressure (TMP) excursion method, feed flow rates and TMP were varied. The results showed a correlation of permeate flux with TMP where the maximum volume concentration factor reached 2.5 times of the initial volume when feed flow rate and TMP were 7 liters/m²/min and 1 bar, respectively. It was optimal operating conditions before TFF system undergone pressure independent regime. In addition, the diafiltration volume was 14 times of the concentrated volume prior to performing a further anion chromatography process.
Platform Development for Vero Cell Culture on Microcarriers Using Dissociation-Reassociation Method
Vero cell is a continuous cell line that is widely used for the production of viral vaccines. However, due to its adherent characteristic, scaling up strategy in large-scale production remains complicated and thus limited. Consequently, suspension-like Vero cell culture processes based on microcarriers have been introduced and employed while also providing increased surface area per volume unit. However, harvesting Vero cells from microcarriers is a huge challenge due to difficulties in cells detaching, lower recovery yield, time-consuming and dissociation agent carry-over. To overcome these problems, we developed a dissociation-association platform technology for detaching and re-attaching cells during subculturing from microcarriers to microcarriers, which will be conveniently applied to seed trains strategies in large scale bioreactors. Herein, Hillex-2 was used to culture Vero cells in serum-containing media using spinner flasks as a scale-down model. The overall confluency of cells on microcarriers was observed using inverted microscope, and the sample cells were daily detached in order to obtain the kinetics data. The metabolites consumption and by-products formation were determined by Nova Biomedical BioprofileFlex.
Establishment and Validation of Correlation Equations to Estimate Volumetric Oxygen Mass Transfer Coefficient (KLa) from Process Parameters in Stirred-Tank Bioreactors Using Response Surface Methodology
Process scale-up is essential for the biological process to increase production capacity from bench-scale bioreactors to either pilot or commercial production. Scale-up based on constant volumetric oxygen mass transfer coefficient (KLa) is mostly used as a scale-up factor since oxygen supply is one of the key limiting factors for cell growth. However, to estimate KLa of culture vessels operated with different conditions are time-consuming since it is considerably influenced by a lot of factors. To overcome the issue, this study aimed to establish correlation equations of KLa and operating parameters in 0.5 L and 5 L bioreactor employed with pitched-blade impeller and gas sparger. Temperature, gas flow rate, agitation speed, and impeller position were selected as process parameters and equations were created using response surface methodology (RSM) based on central composite design (CCD). In addition, the effects of these parameters on KLa were also investigated. Based on RSM, second-order polynomial models for 0.5 L and 5 L bioreactor were obtained with an acceptable determination coefficient (R²) as 0.9736 and 0.9190, respectively. These models were validated, and experimental values showed differences less than 10% from the predicted values. Moreover, RSM revealed that gas flow rate is the most significant parameter while temperature and agitation speed were also found to greatly affect the KLa in both bioreactors. Nevertheless, impeller position was shown to influence KLa in only 5L system. To sum up, these modeled correlations can be used to accurately predict KLa within the specified range of process parameters of two different sizes of bioreactors for further scale-up application.
Eco-Friendly Green Synthesis of Silver Nanoparticles Using Some Egyptian Plants against Some Plant Pathogenic Bacteria and Fungi
Potatoes were the fourth important crop in Egypt which infected by many diseases such as viral, fungal and bacterial diseases. Tubers of sponta were collected from Abo-Homos City, El Behera Governorate, Egypt during season 2014-2015. Bacterial and fungal isolations were performed on the collected samples and the major pathogens were existed in the collected samples are; Fusarium oxisporum (dry rot disease), Ralstonia solaniserum (brown rot disease) and Erwinea carotovora (soft root disease). Molecular identification was performed using 16S r RNA and ITS region for the bacterial and fungal isolates, respectively. The molecular analysis revealed that the obtained sequence showed homology with percentage 99% with the other published strains listed in Gene Bank. Green synthesis of silver nanoparticles using two wild plants, Moringa oliefera (Formulation 1, F1) and Arctostaphylos uvaursi (Formulation 2, F2) as aqueous extracts were performed. The characterization for the biosynthesized nanopartilces was performed using TEM, FTIR, X-RAY diffraction and the results revealed that the obtained nanoparticles are spherical, smooth, and their size ranged from 10 to 50 nm. The activity of the biosynthesised nanoparticles were tested against the three isolated microbes. The results revealed that there is variation in microbe sensitivity obtained by the two nanoparticles formulations with the three different pathogenic microbes. It can be concluded that the nanoparticles have the most potent inhibition effect against bacterial growth at 600 mg/L (F1 and F2) (2.43 and 2.25 cm, respectively) against R. solanacearum. However, P. carotovra was more sensitive than Ralstonia. In addition, the antifungal activity showed that F1 had the highest effect with EC50 value of 999.61 mg/L followed by F2 (EC50 value was 1319.49 mg/L) against F. oxysporium. Nanoparticles synthesized by wild plants could be used as alternative pesticides.
Neuroprotective Effect of Vildagliptin against Cerebral Ischemia in Rats
The burden of stroke is intensely increasing worldwide. Brain injury following transient or permanent focal cerebral ischemia develops ischemic stroke as a consequence of a complex series of pathophysiological events. The aim of this study is to evaluate the possible neuroprotective effect of a dipeptidyl peptidase-4 inhibitor, vildagliptin, independent on its insulinotropic properties in non-diabetic rats subjected to cerebral ischemia. Anaesthetized Wistar rats were subjected to either left middle cerebral artery occlusion (MCAO) or sham operation followed by reperfusion after 30 min of MCAO. The other three groups were orally administered vildagliptin at 3 dose levels (2.5, 5, 10 mg/kg) for 3 successive weeks before subjected to left focal cerebral ischemia/reperfusion and till the end of the study. Neurological deficit scores and motor activity were assessed 24h following reperfusion. 48h following reperfusion, rats were euthanized and their left brain hemispheres were harvested and used in the biochemical, histopathological, and immunohistochemical investigations. Vildagliptin pretreatment improved neurological score deficit, locomotor activity and motor coordination in MCAO rats. Moreover, vildagliptin reduced malondialdehyde (MDA), elevated reduced glutathione (GSH), phosphotylinosital 3 kinase (PI3K), phosphorylated of protein kinase B (p-AKT), and mechanistic target of rapamycin (mTOR) brain contents in addition to reducing protein expression of caspase-3. Also, vildagliptin showed a dose-dependent attenuation in neuronal cell loss and histopathological alterations in MCAO rats. This study proves that vildagliptin exerted the neuroprotective effect in a dose-dependent manner as shown in amelioration of neuronal cell loss and histopathological damage in MCAO rats, which may be mediated by attenuating neuronal and motor deficits, it’s anti-oxidant property, activation of PI3K/AKT/mTOR pathway and its anti-apoptotic effect.
Sustainable Treatment of Vegetable Oil Industry Wastewaters by Xanthomonas campestris
Increasing industrialization as a response to the demands of the consumer society greatly exploits resources and generates large amounts of waste effluents in addition to the desired product. This means it is a priority to implement technologies with the maximum utilization of raw materials and energy, minimum generation of waste effluents and/or their recycling (secondary use). Considering the process conditions and the nature of the raw materials used by the vegetable oil industry, its wastewaters can be used as substrates for the biotechnological production which requires large amounts of water. This way the waste effluents of one branch of industry become raw materials for another branch which produces a new product while reducing wastewater pollution and thereby reducing negative environmental impacts. Vegetable oil production generates wastewaters during the process of rinsing oils and fats which contain mainly fatty acid pollutants. The vegetable oil industry generates large amounts of waste effluents, especially in the processes of degumming, deacidification, deodorization and neutralization. Wastewaters from the vegetable oil industry are generated during the whole year in significant amounts, based on the capacity of the vegetable oil production. There are no known alternative applications for these wastewaters as raw materials for the production of marketable products. Since the literature has no data on the potential negative impact of fatty acids on the metabolism of the bacterium Xanthomonas campestris, these wastewaters were considered as potential raw materials for the biotechnological production of xanthan. In this research, vegetable oil industry wastewaters were used as the basis for the cultivation media for xanthan production with Xanthomonas campestris ATCC 13951. Examining the process of biosynthesis of xanthan on vegetable oil industry wastewaters as the basis for the cultivation media was performed to obtain insight into the possibility of its use in the aforementioned biotechnological process. Additionally, it was important to experimentally determine the absence of substances that have an inhibitory effect on the metabolism of the production microorganism. Xanthan content, rheological parameters of the cultivation media, carbon conversion into xanthan and conversions of the most significant nutrients for biosynthesis (carbon, nitrogen and phosphorus sources) were determined as indicators of the success of biosynthesis. The obtained results show that biotechnological production of the biopolymer xanthan by bacterium Xanthomonas campestris on vegetable oil industry wastewaters based cultivation media simultaneously provides preservation of the environment and economic benefits which is a sustainable solution to the problem of wastewater treatment.
Production, Characterisation, and in vitro Degradation and Biocompatibility of a Solvent-Free Polylactic-Acid/Hydroxyapatite Composite for 3D-Printed Maxillofacial Bone-Regeneration Implants
The current gold-standard for maxillofacial reconstruction surgery (MRS) utilizes auto-grafted cancellous bone as a filler. This study was aimed towards developing a polylactic-acid/hydroxyapatite (PLA-HA) composite suitable for fused-deposition 3D printing. Functionalization of the polymer through the addition of HA was directed to promoting bone-regeneration properties so that the material can rival the performance of cancellous bone grafts in terms of bone-lesion repair. This kind of composite enables the production of MRS implants based off 3D-reconstructions from image studies – namely computed tomography – for anatomically-correct fitting. The present study encompassed in-vitro degradation and in-vitro biocompatibility profiling for 3D-printed PLA and PLA-HA composites. PLA filament (Verbatim Co.) and Captal S hydroxyapatite micro-scale HA powder (Plasma Biotal Ltd) were used to produce PLA-HA composites at 5, 10, and 20%-by-weight HA concentration. These were extruded into 3D-printing filament, and processed in a BFB-3000 3D-Printer (3D Systems Co.) into tensile specimens, and were mechanically challenged as per ASTM D638-03. Furthermore, tensile specimens were subjected to accelerated degradation in phosphate-buffered saline solution at 70°C for 23 days, as per ISO-10993-13-2010. This included monitoring of mass loss (through dry-weighing), crystallinity (through thermogravimetric analysis/differential thermal analysis), molecular weight (through gel-permeation chromatography), and tensile strength. In-vitro biocompatibility analysis included cell-viability and extracellular matrix deposition, which were performed both on flat surfaces and on 3D-constructs – both produced through 3D-printing. Discs of 1 cm in diameter and cubic 3D-meshes of 1 cm3 were 3D printed in PLA and PLA-HA composites (n = 6). The samples were seeded with 5000 MG-63 osteosarcoma-like cells, with cell viability extrapolated throughout 21 days via resazurin reduction assays. As evidence of osteogenicity, collagen and calcium deposition were indirectly estimated through Sirius Red staining and Alizarin Red staining respectively. Results have shown that 3D printed PLA loses structural integrity as early as the first day of accelerated degradation, which was significantly faster than the literature suggests. This was reflected in the loss of tensile strength down to untestable brittleness. During degradation, mass loss, molecular weight, and crystallinity behaved similarly to results found in similar studies for PLA. All composite versions and pure PLA were found to perform equivalent to tissue-culture plastic (TCP) in supporting the seeded-cell population. Significant differences (p = 0.05) were found on collagen deposition for higher HA concentrations, with composite samples performing better than pure PLA and TCP. Additionally, per-cell-calcium deposition on the 3D-meshes was significantly lower when comparing 3D-meshes to discs of the same material (p = 0.05). These results support the idea that 3D-printable PLA-HA composites are a viable resorbable material for artificial grafts for bone-regeneration. Degradation data suggests that 3D-printing of these materials – as opposed to other manufacturing methods – might result in faster resorption than currently-used PLA implants.
Therapeutic Effect of Mango Peel Extract against Local Toxic Effect Induced by Egyptian Echis coloratus Venom
Background: Several million tons of mango peels are considered as annual by-product producing from several factors. In spite of their enrichment with natural products, searching for a new therapeutic effect of mango peel is considered as a promising goal. Aim: The present study was designed to assess both the antioxidant capacity of mango peel extracts (methanolic/ acetone) Hindi cultivar and the neutralizing ability of the extract against the toxic effect of venom of the highest vigorously Egyptian viper; Echis coloratus that may cause both morbidity and mortality. Methods: The total phenolic and flavonoid contents and the antioxidant capacity of the both mango extracts were estimated by free radical scavenging methods (diphenyl-2-picrylhydrazyl; DPPH and azinobis-3-ethylbenzothiazoline-6-sulphonic acid; ABTS ). In addition, the mango extracts were assessed for anti-venom properties under in vitro and in vivo standard assays. Results: In comparison to the methanolic extract, the acetone mango peel extract had the highest total phenolic and flavonoid content at 3.7 ± 0.32 mg of gallic acid equivalent per gram of dry weight (mg GAE/g dw) and 1.55 ± 0.1 mg of catechin equivalent per gram of dry weight (mg CE/g dw), respectively. In addition, the acetone extract showed higher antioxidant potential than the methanolic extract. These findings can be positively correlated to the high content of total polyphenols/flavonoids of the acetone extract. Furthermore, The both extracts were inhibited in vitro the enzymatic activities of protease, L-amino acid oxidase, phospholipase A2 of the venom in a dose-dependent manner. Moreover, Edema, hemorrhage and lethality induced by venom were completely neutralized either by pre-incubation of different doses of the extract with venom or after 10 min of venom injection in the experimental animals. Conclusion: The phenolic compounds of mango peel with potential antioxidant activity may be considered as a new avenue in viper bite therapy.