Search results for: whey protein isolated

Authors: R. Gounina-Allouane, N. Ouali, F. Z. Berrabah, A. Bentaleb

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For a long time, the Gram-positive spore-forming bacteria Bacillus thuringiensis (Bt) has been widely used in biological control against devastating and disease vectors insects. This is due to the insecticidal activity of its crystalline parasporal inclusion (crystals) predominantly comprised of one or more proteins (Cry and Cyt proteins) also called δ-endotoxins, produced during sporulation. The shape and composition of Bt crystals vary among strains and crystalline proteins are extremely varied (more than 475 cry gene were discovered). The insecticidal activity of Bt crystals is very well studied, thus their insecticidal mode of action is well established, however, their antimicrobial effect is largely unknown. The lack of data on the antimicrobial effect of crystalline proteins of Bt and the need for searching new antimicrobial molecules encouraged us to carried out this study. The antibacterial effect of δ-endotoxines produced by two Bt stains; a strain isolated from soil at northern of Algeria (Bt 7.2.B), and a strain isolated from a bioinsecticide (Bacillus thuringiensis var aizawai), activated by proteolysis, was assayed on clinical bacterial strains and ATCC collection ones respectively. Gram positive and negative clinical bacterial strains (Escherichia coli, Klebsiella pneumonaie, Pseudomonas aeruginosa, Staphylococcus aureus) were sensitive to activated Bt 72B endotoxins. Similarly, bacterial strains from ATCC collection (Escherichia coli ATCC 25922, Pseudomonas aerugenosa ATCC 27853, Staphylococcus aureus ATCC 25923) were sensitive to activated B. thuringiensis var aizawai δ-endotoxines. The activated δ-endotoxins were separated by SDS-PAGE.

Keywords: Bacillus thuringiensis, crystals, cry proteins, δ-endotoxins, antibacterial activity

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Authors: Sihem Bazid, Meriem El Kolli, Aicha Medjahed

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Proteins and polysaccharides are the two types of biopolymers most frequently used in the food industry to control the mechanical properties and structural stability and organoleptic properties of the products. The textural and structural properties of these two types of blend polymers depend on their interaction and their ability to form organized structures. From an industrial point of view, a better understanding of mixtures protein / polysaccharide is an important issue since they are already heavily involved in processed food. It is in this context that we have chosen to work on a model system composed of a fibrous protein mixture (gelatin)/anionic polysaccharide (sodium carboxymethylcellulose). Gelatin, one of the most popular biopolymers, is widely used in food, pharmaceutical, cosmetic and photographic applications, because of its unique functional and technological properties. Sodium Carboxymethylcellulose (NaCMC) is an anionic linear polysaccharide derived from cellulose. It is an important industrial polymer with a wide range of applications. The functional properties of this anionic polysaccharide can be modified by the presence of proteins with which it might interact. Another factor may also manage the interaction of protein-polysaccharide mixtures is the triple helix of the gelatin. Its complex synthesis method results in an extracellular assembly containing several levels. Collagen can be in a soluble state or associate into fibrils, which can associate in fiber. Each level corresponds to an organization recognized by the cellular and metabolic system. Gelatin allows this approach, the formation of gelatin gel has triple helical folding of denatured collagen chains, this gel has been the subject of numerous studies, and it is now known that the properties depend only on the rate of triple helices forming the network. Chemical modification of this system is quite controlled. Observe the dynamics of the triple helix may be relevant in understanding the interactions involved in protein-polysaccharides mixtures. Gelatin is central to any industrial process, understand and analyze the molecular dynamics induced by the triple helix in the transitions gelatin, can have great economic importance in all fields and especially the food. The goal is to understand the possible mechanisms involved depending on the nature of the mixtures obtained. From a fundamental point of view, it is clear that the protective effect of NaCMC on gelatin and conformational changes of the α helix are strongly influenced by the nature of the medium. Our goal is to minimize the maximum the α helix structure changes to maintain more stable gelatin and protect against denaturation that occurs during such conversion processes in the food industry. In order to study the nature of interactions and assess the properties of mixtures, polarimetry was used to monitor the optical parameters and to assess the rate of helicity gelatin.

Keywords: gelatin, sodium carboxymethylcellulose, interaction gelatin-NaCMC, the rate of helicity, polarimetry

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Authors: Mi-Hye Hwang, Young Min Son, Kichan Lee, Bang-Hun Hyun, Byeong Yeal Jung

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Botulism is a paralytic disease of human beings and animals caused by neurotoxin produced by Clostridium botulinum. The neurotoxins are genetically distinguished into 8 types, A to H. Ingestion of performed toxin, usually types B, C, and D, have been shown to produce diseases in most cases of cattle botulism. Vaccination is the best measure to prevent cattle botulism. However, the commercially available toxoid-based vaccines are difficult and hazardous to produce. We produced recombinant protein using gene of heavy chain domain of botulinum toxin B of which binds to cellular receptor of neuron cells and used as immunogen. In this study, we evaluated the safety and efficacy of botulism vaccine composed of recombinant types B. Safety test was done by National Regulation for Veterinary Biologicals. For efficacy test, female ICR mice (5 weeks old) were subcutaneously injected, intraperitoneally challenged, and examined the survival rates compared with vaccination and non-vaccination group. Mouse survival rate of recombinant types B vaccine was above 80%, while one of non-vaccination group was 0%. A vaccine composed of recombinant types B was safe and efficacious in mouse. Our results suggest that recombinant heavy chain receptor binding domain can be used as an effective vaccine candidate for type B botulism.

Keywords: botulism, livestock, vaccine, recombinant protein, toxin

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Authors: Noor Khan, Hira Waris

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This study was conducted on genetically male tilapia (GMT) fry reared in glass aquarium for three months to examine the synergetic effect of essential amino acids (EAA) supplementation on growth, body composition, and enzyme activities. Fish having average body weight of 16.56 ± 0.42g were fed twice a day on artificial feed (20% crude protein) procured from Oryza Organics (commercial feed) supplemented with EAA; methionine (M) and lysine (L) designated as T1 (0.3%M and 2%L), T2 (0.6%M and 4%L), T3 (0.9%M and 6%L) and control without EAA. Significantly higher growth performance was observed in T1, followed by T2, T3, and control. The results revealed that whole-body dry matter and crude protein were significantly higher (p ≤ 0.05) in T3 (0.9% and 6%) feeding fish, while the crude fat was lower (p ≤ 0.05) in a similar group of fish. Additionally, protease, amylase, and lipase activities were also observed maximum (p ≤ 0.05) in response to T3 than other treatments and control. However, the EAA, especially lysine and methionine, were found significantly higher (p ≤ 0.05) in T1 compared to other treatments. Conclusively, the addition of EAA, methionine, and lysine in the feed not only enhanced the growth performance of GMT fry but also improved body proximate composition and essential amino acid profile.

Keywords: genetically male tilapia, body composition, digestive enzyme activities, amino acid profile

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Authors: Yohanes Buang, Suwari

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Increasing interest of society in use and creation of herbal medicines has encouraged scientists/researchers to establish an ideal method to produce the best quality and quantity of pharmaceutical extracts. To have highest the antioxidative extracts, the method used must be at optimum conditions. Hence, the best method is not only able to provide highest quantity and quality of the isolated pharmaceutical extracts but also it has to be easy to do, simple, fast, and cheap. The characterization of solvents in maceration technique, in present study, involved various variables influencing quantity and quality of the pharmaceutical extracts, such as solvent’s optimum acidity-alkalinity (pH), temperature, concentration, and contact time. The shifting polarity of the solvent by combinations of water with ethanol (70:30) and (50:50) were also performed to completely record the best solvent system in application of maceration technology. Among those three solvents threated within Myrmecodia pendens, as a model of natural product, the results showed that water solvent system with conditions of alkalinity pH, optimum temperature, concentration, and contact time, is the best system to perform the maceration in order to have the highest isolated antioxidative activated extracts. The optimum conditions of the water solvent are at the alkalinity pH 9 up, 30 mg/mL of concentration, 40 min of contact time, 100 °C of temperature, and no ethanol used to replace parts of the water solvent. The present study strongly recommended the best conditions of solvent system to isolate the pharmaceutical extracts of natural products in application of the maceration technology.

Keywords: extracts, herbal medicine, natural product, maceration technique

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Authors: Indu Bhushan Sharma, Rashmi Saraswat

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The thermotolerant, solvent stable and alkalophilic lipase producing bacterial strain was isolated from the water sample of the foothills of Trikuta Mountain in Kakryal (Reasi district) in Jammu and Kashmir, India. The lipase-producing microorganisms were screened using tributyrin agar plates. The selected microbe was optimized for maximum lipase production by subjecting to various carbon and nitrogen sources, incubation period and inoculum size. The selected strain was identified as Bacillus subtilis strain kakrayal_1 (BSK_1) using 16S rRNA sequence analysis. Effect of pH, temperature, metal ions, detergents and organic solvents were studied on lipase activity. Lipase was found to be stable over a pH range of 6.0 to 9.0 and exhibited maximum activity at pH 8. Lipolytic activity was highest at 37°C and the enzyme activity remained at 60°C for 24hrs, hence, established as thermo-tolerant. Production of lipase was significantly induced by vegetable oil and the best nitrogen source was found to be peptone. The isolated Bacillus lipase was stimulated by pre-treatment with Mn2+, Ca2+, K+, Zn2+, and Fe2+. Lipase was stable in detergents such as triton X 100, tween 20 and Tween 80. The 100% ethyl acetate enhanced lipase activity whereas, lipase activity were found to be stable in Hexane. The optimization resulted in 4 fold increase in lipase production. Bacillus lipases are ‘generally recognized as safe’ (GRAS) and are industrially interesting. The inducible alkaline, thermo-tolerant lipase exhibited the ability to be stable in detergents and organic solvents. This could be further researched as a potential biocatalyst for industrial applications such as biotransformation, detergent formulation, bioremediation and organic synthesis.

Keywords: bacillus, lipase, thermotolerant, alkalophilic

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Authors: Joshua Bradley, Matthew Buns

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This study aimed to identify the effect of concurrent nutritional supplementation on soccer performance as players ingested either carbohydrate CHO (52 g of Cytocarb Maltodextrin) or a combined carbohydrate and protein PRO (Muscle Milk Pro Series 17g CHO + 50 g PRO liquid) supplement. Twelve male, junior college soccer players (age: 18 ± 6 years, wt. 73.3 ± 8.6 kg) completed three trials wearing global positioning systems (GPS) to measure total running distance and sprinting distance during soccer simulation games. The first match simulation was a baseline match with no supplementation. One hour prior to the second match, simulation players were randomly assigned to one of two supplemental groups CHO or CHO + PRO. A repeated measures ANOVA with a Greenhouse-Geisser correction revealed a statistically significant increase in the total distance run for the CHO supplementation group in comparison to the CHO + PRO group (10.19 ± .200 km vs. 9.77± .194km, p = .035). Although the total running distance was meaningfully influenced by the supplementation, the pattern of response for total sprinting distance was not influenced by supplementation. There was a decline in sprinting distance and total running distance from first half to second half, both for the control (M = -0.01 km, SD = 0.17) and CHO supplementation group (-0.04 km, SD = .19), although these differences were not statistically meaningful. There was a positive correlation between sprinting distance and total distance, which was statistically significant (r = -.514, n = 36, p = .01) In conclusion, supplementation influenced the pattern of activity and demonstrated between-trial differences.

Keywords: GPS, nutrition, simulation, supplementation

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Authors: Gadiri Sabiha

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Introduction : IgE-dependent cow's milk protein allergy (APLV) is one of the most common allergies in children and is one of the three most common allergies observed in children under 6 years of age. Its natural evolution is most often towards healing. The objective is to determine the sensitization profile of patients allergic to cow's milk (VL). Material and method :A retrospective study carried out on a pediatric population (age < 12 years) over a period of four years (2018-2021) in the context of a suspected food allergy to cow's milk proteins carried out on 121 children aged between 8 months -12 years The search for specific IgE was carried out by immunodot (EUROLINE Pediatric; EUROIMMUN) test which allows a semi-quantitative determination of specific IgE. Results 36 patients (29.7%) had a cow's milk protein allergy (ALPV) with a slight female predominance (58.33% girls vs 41.66% boys) The main clinical signs were: acute diarrhoea; vomiting; Intense abdominal pain, and cutaneous signs (pruritus/urticaria) with respective frequencies of 72%; 58%; 44% and 19%. The 3 major and specific VL allergens identified were beta-lactoglobulin 59% caseins 51% and alpha-lactalbumin 29.7%, The profile of sensitization to LV varies according to age, in infants before 1 year of anti-casein, IgE are predominant 83.3%, followed by beta-lactoglobulin 66.66% and alpha-lactolbumin 50% Conclusion CMPA is a frequent pathology which ranks among the three most common food allergies in children. This is the first to appear, most often starting in infants under 6 months old.

Keywords: specific Ige, food allergy, cow 's milk, child

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Authors: Dina Fatmawati, Sofia Mubarika, Mae Sri Wahyuningsih

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Chemotherapy for breast cancer is largely ineffective, but innovative combinations of chemotherapeutic agents and natural compounds represent a promising strategy. In our previous study, the combination of Doxorubicin (Dox) and ethanolic extract of Dioscorea esculenta tuber ((EED) was found to have a synergistic effect on T47D human breast cancer cell line. In this study, we investigated the apoptotic effect of the combination on T47D human breast cancer cells and normal fibroblasts cell line and its effects on the expression of Caspase-3 and cleaved poly (ADP-Ribose) Polymerase-1 (cPARP-1) protein. T47D cell lines and fibroblasts cells were treated with the combination of Dox and EED. Apoptotic effect of the combination was determined using flow cytrometry assay. Protein expressions were determined by immunocytochemistry staining. The percentage of apoptotic cells were significantly higher in T47D cell lines (75%) than that of in fibroblast cells (23%). The expression of Caspase 3 (84.53%) and cPARP-1 (83.36%) were significantly higher in the cancer cell lines than those of normal cells. These results indicate that the combination of doxorubicin and Dioscorea esculenta is a promising candidate for the treatment of breast cancer cells.

Keywords: Dioscorea esculenta, Doxorubicin, apoptosis, immunocytochemistry, cancer cells

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Authors: Syed Asif Hassan, Tabrej Khan

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Tuberculosis (TB) caused by bacillus Mycobacterium tuberculosis (Mtb) continues to take a disturbing toll on human life and healthcare facility worldwide. The global burden of TB remains enormous. The alarming rise of multi-drug resistant strains of Mycobacterium tuberculosis calls for an increase in research efforts towards the development of new target specific therapeutics against diverse strains of M. tuberculosis. Therefore, the discovery of new molecular scaffolds targeting new drug sites should be a priority for a workable plan for fighting resistance in Mycobacterium tuberculosis (Mtb). Mtb non-acylated lipoprotein LprG (Rv1411c) has a Toll-like receptor 2 (TLR2) agonist actions that depend on its association with triacylated glycolipids binding specifically with the hydrophobic pocket of Mtb LprG lipoprotein. The detection of a glycolipid carrier function has important implications for the role of LprG in Mycobacterial physiology and virulence. Therefore, considering the pivotal role of glycolipids in mycobacterial physiology and host-pathogen interactions, designing competitive antagonist (chemotherapeutics) ligands that competitively bind to glycolipid binding domain in LprG lipoprotein, will lead to inhibition of tuberculosis infection in humans. In this study, a unified approach involving ligand-based virtual screening protocol USRCAT (Ultra Shape Recognition) software and molecular docking studies using Auto Dock Vina 1.1.2 using the X-ray crystal structure of Mtb LprG protein was implemented. The docking results were further confirmed by DSX (DrugScore eXtented), a robust program to evaluate the binding energy of ligands bound to the Ligand binding domain of the Mtb LprG lipoprotein. The ligand, which has the higher hypothetical affinity, also has greater negative value. Based on the USRCAT, Lipinski’s values and molecular docking results, [(2R)-2,3-di(hexadecanoyl oxy)propyl][(2S,3S,5S,6R)-3,4,5-trihydroxy-2,6-bis[[(2R,3S,4S,5R,6S)-3,4,5-trihydroxy-6 (hydroxymethyl)tetrahydropyran-2-yl]oxy]cyclohexyl] phosphate (XPX) was confirmed as a promising drug-like lead compound (antagonist) binding specifically to the hydrophobic domain of LprG protein with affinity greater than that of PIM2 (agonist of LprG protein) with a free binding energy of -9.98e+006 Kcal/mol and binding affinity of -132 Kcal/mol, respectively. A further, in vitro assay of this compound is required to establish its potency in inhibiting molecular evasion mechanism of MTB within the infected host macrophages. These results will certainly be helpful in future anti-TB drug discovery efforts against Multidrug-Resistance Tuberculosis (MDR-TB).

Keywords: antagonist, agonist, binding affinity, chemotherapeutics, drug-like, multi drug resistance tuberculosis (MDR-TB), RV1411c protein, toll-like receptor (TLR2)

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Authors: Nandhana Vivek, Bhaskar Gogoi, Ayyavu Mahesh

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Breast cancer metastasis plays a key role in cancer progression and fatality. The present study examines the potential causes of metastasis in breast cancer by investigating the novel interactions between genes and their pathways. The gene expression profile of GSE99394, GSE1246464, and GSE103865 was downloaded from the GEO data repository to analyze the differentially expressed genes (DEGs). Protein-protein interactions, target factor interactions, pathways and gene relationships, and functional enrichment networks were investigated. The proliferation pathway was shown to be highly expressed in breast cancer progression and metastasis in all three datasets. Gene Ontology analysis revealed 11 DEGs as gene targets to control breast cancer metastasis: LYN, DLGAP5, CXCR4, CDC6, NANOG, IFI30, TXP2, AGTR1, MKI67, and FTH1. Upon studying the function, genomic and proteomic data, and pathway involvement of the target genes, DLGAP5 proved to be a promising candidate due to it being highly differentially expressed in all datasets. The study takes a unique perspective on the avenues through which DLGAP5 promotes metastasis. The current investigation helps pave the way in understanding the role DLGAP5 plays in metastasis, which leads to an increased incidence of death among breast cancer patients.

Keywords: genomics, metastasis, microarray, cancer

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Authors: Bhavini Patel, Aslihan Ugun-Klusek, Ellen Billet

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The two main contributing factors to familial and idiopathic form of Parkinson’s disease (PD) are oxidative stress and altered proteolysis. Monoamine oxidase-A (MAO-A) plays a significant role in redox homeostasis by producing reactive oxygen species (ROS) via deamination of for example, dopamine. The ROS generated induces chemical modification of proteins resulting in altered biological function. The ubiquitin-proteasome system, which consists of three different types or proteolytic activity, namely “chymotrypsin-like” activity (CLA), “trypsin-like” activity (TLA) and “post acidic-like” activity (PLA), is responsible for the degradation of ubiquitinated proteins. Defects in UPS are known to be strongly correlated to PD. Herein, the effect of ROS generated by MAO-A on proteasome activity and the effects of proteasome inhibition on MAO-A protein levels in WT, mock and MAO-A overexpressed (MAO-A+) SHSY5Y neuroblastoma cell lines were investigated. The data in this study report increased proteolytic activity when MAO-A protein levels are significantly increased, in particular CLA and PLA. Additionally, 20S proteasome inhibition induced a decrease in MAO-A levels in WT and mock cells in comparison to MAO-A+ cells in which 20S proteasome inhibition induced increased MAO-A levels to be further increased at 48 hours of inhibition. This study supports the fact that MAO-A could be a potential pharmaceutical target for neuronal protection as data suggests that endogenous MAO-A levels may be essential for modulating cell death and survival.

Keywords: monoamine oxidase, neurodegeneration, Parkinson's disease, proteasome

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Authors: Alok Kumar, Hari Ram, Lebin Thomas, Ved Pal Singh

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Organic solvent tolerant amylases and proteases of microbial origin are in great demand for their application in transglycosylation of water-insoluble flavanoids and in peptide synthesizing reaction in organic media. Most of the amylases and proteases are unstable in presence of organic solvent. In the present work two different bacterial strains M-11 and VP-07 were isolated from the soil sample of a dairy farm in Delhi, India, for the efficient production of extracellular amylase and protease through their screening on starch agar (SA) and skimmed milk agar (SMA) plates, respectively. Both the strains (M-11 and VP-07) were identified based on morphological, biochemical and 16S rRNA gene sequencing methods. After analysis through Ez-Taxon software, the strains M-11 and VP-07 were found to have maximum pairwise similarity of 98.63% and 100% with Bacillus subtilis subsp. inaquosorum BGSC 3A28 and Bacillus anthracis ATCC 14578 and were therefore identified as Bacillus sp. UKS1 and Bacillus sp. UKS2, respectively. Time course study of enzyme activity and bacterial growth has shown that both strains exhibited typical sigmoid growth behavior and maximum production of amylase (180 U/ml) and protease (78 U/ml) by these strains (UKS1 and UKS2) was commenced during stationary phase of growth at 24 and 20 h, respectively. Thereafter, both amylase and protease were tested for their tolerance towards organic solvents and were found to be active as well stable in p-xylene (130% and 115%), chloroform (110% and 112%), isooctane (119% and 107%), benzene (121% and 104%), n-hexane (116% and 103%) and toluene (112% and 101%, respectively). Owing to such properties, these enzymes can be exploited for their potential application in industries for organic synthesis.

Keywords: amylase, enzyme activity, industrial applications, organic solvent tolerant, protease

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Authors: Mehmet Demir Kaya, Engin Gökhan Kulan, Onur İleri, Süleyman Avcı

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This research was conducted to determine the difference in seed vigor between the seed lots cultivated in main and second crop of soybean in Turkey. Seeds from soybean cv. Cinsoy and Umut-2002 were evaluated in the laboratory for germination, emergence, cool test at 18°C for 10 days, and cold test at 10°C for 4 days and 25°C for 6 days. Result showed that the initial oil contents of Cinsoy and Umut-2002 and seeds were determined to be 19.8 and 20.1% in main crop, and 18.7 and 22.1% in second crop, respectively. It was determined that a clear difference between main and second crop soybean seed lots for seed vigor was found. Germination and emergence percentage were higher in the seed from second crop cultivation of the cultivars. There was no significant difference in germination percentage in cool and cold test while seedling growth was better in the seeds of second crop soybean. The highest seed vigor index (477.6) was found in the seeds of the cultivars grown at second crop. Standard germination percentage did not give a sensitive separation for determining seed vigor of soybean lots. It was concluded that second crop soybean seeds were found the most suitable for seed production while main crop soybean gave higher protein lower oil content.

Keywords: Glycine max L., germination, emergence, protein content, vigor test

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Authors: C. Benoudjafer, M. N. Tandjaoui, C. Benachaiba

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The use of kinetic energy of the wind is in full rise in the world and it starts to be known in our country but timidly. One or more aero generators can be installed to produce for example electricity on isolated places or not connected to the electrical supply network. To use the wind as energy source, it is necessary to know first the energy needs for the population and study the wind intensity, speed, frequency and direction.

Keywords: Algeria, renewable energies, wind, wind power, aero-generators, wind energetic potential

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Authors: A. Ferasat, S. Rostampour Yasouri

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Seasonal flu is a highly contagious infection caused by influenza viruses. These viruses undergo genetic changes that result in new epidemics across the globe. Medical attention is crucial in severe cases, particularly for the elderly, frail, and those with chronic illnesses, as their immune systems are often weaker. The purpose of this study was to detect new subtypes of the influenza A virus rapidly using a specific RT-PCR method based on the HA gene (hemagglutinin). In the winter and spring of 2022_2023, 120 embryonated egg samples were cultured, suspected of seasonal influenza. RNA synthesis, followed by cDNA synthesis, was performed. Finally, the PCR technique was applied using a pair of specific primers designed based on the HA gene. The PCR product was identified after purification, and the nucleotide sequence of purified PCR products was compared with the sequences in the gene bank. The results showed a high similarity between the sequence of the positive samples isolated from the patients and the sequence of the new strains isolated in recent years. This RT-PCR technique is entirely specific in this study, enabling the detection and multiplication of influenza and its subspecies from clinical samples. The RT-PCR technique based on the HA gene, along with sequencing, is a fast, specific, and sensitive diagnostic method for those infected with influenza viruses and its new subtypes. Rapid molecular diagnosis of influenza is essential for suspected people to control and prevent the spread of the disease to others. It also prevents the occurrence of secondary (sometimes fatal) pneumonia that results from influenza and pathogenic bacteria. The critical role of rapid diagnosis of new strains of influenza is to prepare a drug vaccine against the latest viruses that did not exist in the community last year and are entirely new viruses.

Keywords: influenza, molecular diagnosis, patients, RT-PCR technique

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Authors: Rachel Karasenty Saltoun, Charlotte Roddick, Chelsea D. Christie, Frances Chen

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Moving has become an integral part of many people’s lives. This research explores whether relocating to a new city is associated with an increase in loneliness and cardiovascular disease risk and if this increased risk diminishes with continued residency. To test this, various psychosocial variables and three cardiovascular disease risk markers (C-reactive protein, albumin, blood pressure) were assessed on two groups of individuals: those who have moved to Vancouver, Canada in the previous 6 weeks (‘Movers’) and those who have lived in Vancouver for at least five years (‘Non-Movers’). It was hypothesized that individuals who had recently relocated would have heightened levels of loneliness, blood pressure (BP), albumin, and C-reactive protein (CRP) compared to those who had not recently relocated. Length of residency was hypothesized to moderate these effects, such that after a few months, loneliness levels and cardiovascular disease risk would decrease among those who had recently relocated. Correlational analysis indicated a trend between the change in CRP and albumin levels and loneliness overtime on an individual level. However, these results must be interpreted with caution due to the small sample size. As Vancouver’s immigration rates continue to grow, this study has important implications regarding the social support resources offered to new immigrants, as well as bringing awareness at the healthcare level of the potential increase in cardiovascular disease risk among those who have recently relocated.

Keywords: cardiovascular disease risk, loneliness, moving, residential mobility

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Authors: Jerman Madonsela, Wallace Matizamhuka, Akiko Yamamoto, Ronald Machaka, Brendon Shongwe

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In this study, biocompatibility evaluation of nanostructured near beta Ti-24Nb-4Zr-8Sn (Ti2448) alloy with non-toxic elements produced utilizing Spark plasma sintering (SPS) of very fine microsized powders attained through mechanical alloying was performed. The results were compared with pure titanium and Ti-6Al-4V (Ti64) alloy. Cell proliferation test was performed using murine osteoblastic cells, MC3T3-E1 at two cell densities; 400 and 4000 cells/mL for 7 days incubation. Pure titanium took a lead under both conditions suggesting that the presence of other oxide layers influence cell proliferation. No significant difference in cell proliferation was observed between Ti64 and Ti2448. Potentiodynamic measurement in Hanks, 0.9% NaCl and cell culture medium showed no distinct difference on the anodic polarization curves of the three alloys, indicating that the same anodic reaction occurred on their surface but with different rates. However, Ti2448 showed better corrosion resistance in cell culture medium with a slightly lower corrosion rate of 2.96 nA/cm2 compared to 4.86 nA/cm2 and 5.62 nA/cm2 of Ti and Ti64 respectively. Ti2448 adsorbed less protein as compared to Ti and Ti64 though no notable difference in surface wettability was observed.

Keywords: biocompatibility, osteoblast, corrosion, surface wettability, protein adsorption

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Authors: Ramiro Quintanilla-Licea, Isvar K. Angeles-Hernandez, Javier Vargas-Villarreal

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Amebiasis caused by Entamoeba histolytica, associated with high morbidity and mortality, is currently a significant public health problem worldwide, especially in developing countries. In the world, around 50 million people develop this illness, and up to 100,000 deaths occur annually. Due to the side-effects and the resistance that pathogenic protozoa show against common antiparasitic drugs (e.g., metronidazole), growing attention has been paid to plants used in traditional medicine around the world to find new antiprotozoal agents. In this study is reported about the isolation and structure elucidation of antiamoebic compounds occurring in Lippia graveolens Kunth (Mexican oregano). The work-up of the methanol extract of L. graveolens afforded the known flavonoids pinocembrin (1), sakuranetin (2), cirsimaritin (3) and naringenin (4) by bioguided isolation using several chromatographic techniques. Structural elucidation of the isolated compounds was based on spectroscopic/spectrometric analyses (IR; 1H- and 13C-NMR; MS) and comparison with literature data. These compounds showed significant antiprotozoal activity against Entamoeba histolytica trophozoites using in vitro tests (positive control metronidazole IC50 0.205 µg/mL). The antiprotozoal activity of pinocembrin and naringenin (IC50 of 29.51 µg/mL and 28.85 µg/mL, respectively) was higher compared with sakuranetin (44.47 µg/mL) and with cirsimaritin (150.00 µg/mL), revealing that a 5,7-dihydroxylated A ring is essential for antiprotozoal activity. These research funds may validate the use of this plant in the traditional Mexican medicine for the treatment of some digestive disorders and can help to integrate the use of extracts of L. graveolens in the conventional and complementary medicine for the treatment of parasitic diseases.

Keywords: amoebiasis, antiprotozoal agents, bioguided isolation, infectious diseases

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Authors: Saroj Poudel, Joshua Mancini, Douglas Pike, Jennifer Timm, Alexei Tyryshkin, Vikas Nanda, Paul Falkowski

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On the early Earth, protein-metal complexes likely harvested energy from a reduced environment. These complexes would have been precursors to the metabolic enzymes of ancient organisms. Hydrogenase is an essential enzyme in most anaerobic organisms for the reduction and oxidation of hydrogen in the environment and is likely one of the earliest evolved enzymes. To attempt to reinvent a precursor to modern hydrogenase, we computationally designed a short thirteen amino acid peptide that binds the often-required catalytic transition metal Nickel in hydrogenase. This simple complex can achieve hundreds of hydrogen evolution cycles using light energy in a broad range of temperature and pH. Biophysical and structural investigations strongly indicate the peptide forms a di-nickel active site analogous to Acetyl-CoA synthase, an ancient protein central to carbon reduction in the Wood-Ljungdahl pathway and capable of hydrogen evolution. This work demonstrates that prior to the complex evolution of multidomain enzymes, early peptide-metal complexes could have catalyzed energy transfer from the environment on the early Earth and enabled the evolution of modern metabolism

Keywords: hydrogenase, prebiotic enzyme, metalloenzyme, computational design

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Authors: Nwadike Emmanuel Chinagoron, Achebe Chukwunonso, Ezeliora Chukwuemeka Daniel, Azaka Onyemazuwa Andrew

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In this research work, the process of biodiesel production in a pilot plant was studied using cow tallow as raw material, methanol as the solvent and potassium hydroxide as catalysts. The biodiesel quality was determined by characterization. The tallow used in the production had a molecular weight of 860g. Its oil had a density value of 0.8g/ml, iodine value of 63.45, viscosity at 300C was 9.83pas, acid value was 1.96, free fatty acid (FFA) of 0.98%, saponification value of 82.75mleq/kg, specific gravity of 0.898, flash point of 1100C, cloud point of 950C and Calorific value also called Higher Heating Value (HHV) of 38.365MJ/Kg. The produced biodiesel had a density of 0.82g/ml, iodine value of 126.9, viscosity of 4.32pas at 300C, acid value of 0.561, FFA of 0.2805%, saponification value of 137.45 mleq/kg.Flash point, cloud point and centane number of the biodiesel produced are 1390C, 980C and 57.5 respectively, with fat content, protein content, ash content, moisture content, fiber content and carbohydrate content values of 10%, 2.8%, 5%, 5%, 20%, and 37.2% respectively. The biodiesel higher heating values (calorific values) when estimated from viscosity, density and flash points were 41.4MJ/Kg, 63.8MJ/Kg, and 34.6MJ/Kg respectively. The biodiesel was blended with conventional diesel. The blend B-10 had values of 1320C and 960C for flash and cloud points, with Calorific value (or HHV) of 34.6 MJ/Kg (when estimated from its Flash point) and fat content, protein content, ash content, moisture content, fiber content and carbohydrate content values of 5%, 2.1%,10%, 5%, 15%, and 62.9% respectively.

Keywords: biodiesel, characterization, cow-tallow, cetane rating

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Authors: Marthese Azzopardi, Liberato Camilleri

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This research was performed to investigate the study habits and level of difficulty perceived by post-secondary students in Biology at Advanced-level topics after completing their first year of study. At the end of a two-year ‘sixth form’ course, Maltese students sit for the Matriculation and Secondary Education Certificate (MATSEC) Advanced-level biology exam as a requirement to pursue science-related studies at the University of Malta. The sample was composed of 23 students (16 taking Chemistry and seven taking some ‘Other’ subject at the Advanced Level). The cohort comprised seven males and 16 females. A questionnaire constructed by the authors, was answered anonymously during the last lecture at the end of the first year of study, in May 2016. The Chi square test revealed that gender plays no effect on the various study habits (c² (6) = 5.873, p = 0.438). ‘Reading both notes and textbooks’ was the most common method adopted by males (71.4%), whereas ‘Writing notes on each topic’ was that mostly used by females (81.3%). The Mann-Whitney U test showed no significant difference in the study habits of students and the mean assessment mark obtained at the end of the first year course (p = 0.231). Statistical difference was found with the One-ANOVA test when comparing the mean assessment mark obtained at the end of the first year course when students are clustered by their Secondary Education Certificate (SEC) grade (p < 0.001). Those obtaining a SEC grade of 2 and 3 got the highest mean assessment of 68.33% and 66.9%, respectively [SEC grading is 1-7, where 1 is the highest]. The Friedman test was used to compare the mean difficulty rating scores provided for the difficulty of each topic. The mean difficulty rating score ranges from 1 to 4, where the larger the mean rating score, the higher the difficulty. When considering the whole group of students, nine topics out of 21 were perceived as significantly more difficult than the other topics. Protein synthesis, DNA Replication and Biomolecules were the most difficult, in that order. The Mann-Whitney U test revealed that the perceived level of difficulty in comprehending Biomolecules is significantly lower for students taking Chemistry compared to those not choosing the subject (p = 0.018). Protein Synthesis was claimed as the most difficult by Chemistry students and Biomolecules by those not studying Chemistry. DNA Replication was the second most difficult topic perceived by both groups. The Mann-Whitney U test was used to examine the effect of gender on the perceived level of difficulty in comprehending various topics. It was found that females have significantly more difficulty in comprehending Biomolecules than males (p=0.039). Protein synthesis was perceived as the most difficult topic by males (mean difficulty rating score = 3.14), while Biomolecules, DNA Replication and Protein synthesis were of equal difficulty for females (mean difficulty rating score = 3.00). Males and females perceived DNA Replication as equally difficult (mean difficulty rating score = 3.00). Discovering the students’ study habits and perceived level of difficulty of specific topics is vital for the lecturer to offer guidance that leads to higher academic achievement.

Keywords: biology, perceived difficulty, post-secondary, study habits

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Authors: Musarat Ishaq, Tara Karnezis, Ramin Shayan

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Lipedema, a poorly understood chronic disease of adipose hyper-deposition, is often mistaken for obesity and causes significant impairment to mobility and quality-of-life. To identify molecular mechanisms underpinning lipedema, we employed comprehensive omics-based comparative analyses of whole tissue, adipocyte precursors (adipose-derived stem cells (ADSCs)), and adipocytes from patients with or without lipedema. Transcriptional profiling revealed significant differences in lipedema tissue, adipocytes, and ADSCs, with altered levels of mRNAs involved inproliferation and cell adhesion. One highly up-regulated gene in lipedema adipose tissue, adipocytes and ADSCs, ZIC4, encodes Zinc Finger Protein ZIC 4, a class of transcription factor which may be involved in regulating metabolism and adipogenesis. ZIC4 inhibition impaired the adipogenesis of ADSCs into mature adipocytes. Epigenetic regulation study revealed overexpression of ZIC4 is involved in decreased promoter DNA methylation and subsequent decrease in adipogenesis. These epigenetic modifications can alter adipocytes microenvironment and adipocytes differentiation. Our study show that epigenetic events regulate the ability of ADSCs to commit and differentiate into mature adipocytes by modulating ZIC4.

Keywords: lipedema, adipose-derived stem cells, adipose tisue, adipocytes, zinc finger protein, epigenetic

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Authors: Syeda Kiran Shahzadi, Nasir Mahmood, Muhammad Abdul Qadir

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In the current research, three recombinant human interferon alpha-2b proteins (two modified and one normal form) were produced and Pegylated with an aim to produce more effective drugs against viral infections and cancers. The modified recombinant human interferon alpha-2b proteins were produced by site-directed modifications of interferon alpha 2b gene, targeting the amino acids at positions ‘R23’ and ‘H34’. The resulting chemically modified and unmodified forms of human interferon alpha 2b were conjugated with methoxy-polyethylene glycol propanealdehyde (400 KDa) and methoxy-polyethylene glycol succinimidyl succinate (400 KDa). Pegylation of normal and modified forms of Interferon alpha-2b prolong their release time and enhance their efficacy. The conjugation of PEG with modified and unmodified human interferon alpha 2b protein drugs was also characterized with 1H-NMR, HPLC, and SDS-PAGE. Antiproliferative assays of modified and unmodified forms of drugs were performed in cell based bioassays using MDBK cell lines. The results indicated that experimentally produced recombinant human interferon alpha-2b proteins were biologically active and resulted in significant inhibition of cell growth.

Keywords: protein refolding, antiproliferative activities, biomedical applications, human interferon alpha-2b, pegylation, mPEG-propionaldehyde, site directed mutagenesis, E. coli expression

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Authors: Chu Wan-Loy, Kok Yih-Yih, Choong Siew-Ling

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The human health risks due to poor air quality caused by a wide array of microorganisms have attracted much interest. Airborne algae have been reported as early as 19th century and they can be found in the air of tropic and warm atmospheres. Airborne algae normally originate from water surfaces, soil, trees, buildings and rock surfaces. It is estimated that at least 2880 algal cells are inhaled per day by human. However, there are relatively little data published on airborne algae and its related adverse health effects except sporadic reports of algae associated clinical allergenicity. A collection of airborne algae cultures has been established following a recent survey on the occurrence of airborne algae in indoor and outdoor environments in Kuala Lumpur. The aim of this study was to investigate the allergenic potential of the isolated airborne green and blue-green algae, namely Scenedesmus sp., Cylindrospermum sp. and Hapalosiphon sp.. The suspensions of freeze-dried airborne algae were adminstered into balb-c mice model through intra-nasal route to determine their allergenic potential. Results showed that Scenedesmus sp. (1 mg/mL) increased the systemic Ig E levels in mice by 3-8 fold compared to pre-treatment. On the other hand, Cylindrospermum sp. and Hapalosiphon sp. at similar concentration caused the Ig E to increase by 2-4 fold. The potential of airborne algae causing Ig E mediated type 1 hypersensitivity was elucidated using other immunological markers such as cytokine interleukin (IL)- 4, 5, 6 and interferon-ɣ. When we compared the amount of interleukins in mouse serum between day 0 and day 53 (day of sacrifice), Hapalosiphon sp. (1mg/mL) increased the expression of IL4 and 6 by 8 fold while the Cylindrospermum sp. (1mg/mL) increased the expression of IL4 and IFɣ by 8 and 2 fold respectively. In conclusion, repeated exposure to the three selected airborne algae may stimulate the immune response and generate Ig E in a mouse model.

Keywords: airborne algae, respiratory, allergenic, immune response, Malaysia

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Authors: Joanna Gerszon, Aleksandra Rodacka

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In the pathogenesis of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease, protein and peptide aggregation processes play a vital role in contributing to the formation of intracellular and extracellular protein deposits. One of the major components of these deposits is the oxidatively modified glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Therefore, the purpose of this research was to answer the question whether piceatannol, a stilbene derivative, counteracts and/or slows down oxidative stress-induced GAPDH aggregation. The study also aimed to determine if this natural occurring compound prevents unfavorable nuclear translocation of GAPDH in hippocampal cells. The isothermal titration calorimetry (ITC) analysis indicated that one molecule of GAPDH can bind up to 8 molecules of piceatannol (7.3 ± 0.9). As a consequence of piceatannol binding to the enzyme, the loss of activity was observed. Parallel with GAPDH inactivation the changes in zeta potential, and loss of free thiol groups were noted. Nevertheless, the ligand-protein binding does not influence the secondary structure of the GAPDH. Precise molecular docking analysis of the interactions inside the active center allowed to presume that these effects are due to piceatannol ability to assemble a covalent binding with nucleophilic cysteine residue (Cys149) which is directly involved in the catalytic reaction. Molecular docking also showed that simultaneously 11 molecules of ligand can be bound to dehydrogenase. Taking into consideration obtained data, the influence of piceatannol on level of GAPDH aggregation induced by excessive oxidative stress was examined. The applied methods (thioflavin-T binding-dependent fluorescence as well as microscopy methods - transmission electron microscopy, Congo Red staining) revealed that piceatannol significantly diminishes level of GAPDH aggregation. Finally, studies involving cellular model (Western blot analyses of nuclear and cytosolic fractions and confocal microscopy) indicated that piceatannol-GAPDH binding prevents GAPDH from nuclear translocation induced by excessive oxidative stress in hippocampal cells. In consequence, it counteracts cell apoptosis. These studies demonstrate that by binding with GAPDH, piceatannol blocks cysteine residue and counteracts its oxidative modifications, that induce oligomerization and GAPDH aggregation as well as it prevents hippocampal cells from apoptosis by retaining GAPDH in the cytoplasm. All these findings provide a new insight into the role of piceatannol interaction with GAPDH and present a potential therapeutic strategy for some neurological disorders related to GAPDH aggregation. This work was supported by the by National Science Centre, Poland (grant number 2017/25/N/NZ1/02849).

Keywords: glyceraldehyde-3-phosphate dehydrogenase, neurodegenerative disease, neuroprotection, piceatannol, protein aggregation

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Authors: Ashish Kumar Agrahari, Amit Kumar

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Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired clonal blood disorder that manifests with hemolytic anemia, thrombosis, and peripheral blood cytopenias. The disease is caused by the deficiency of two glycosylphosphatidylinositols (GPI)-anchored proteins (CD55 and CD59) in the hemopoietic stem cells. The deficiency of GPI-anchored proteins has been associated with the somatic mutations in phosphatidylinositol glycan class A (PIGA). However, the mutations that do not cause PNH is associated with the multiple congenital anomalies-hypotonia-seizures syndrome 2 (MCAHS2). To best of our knowledge, no computational study has been performed to explore the atomistic level impact of PIGA mutations on the structure and dynamics of the protein. In the current work, we are mainly interested to get insights into the molecular mechanism of PIGA mutations. In the initial step, we screened the most pathogenic mutations from the pool of publicly available mutations. Further, to get a better understanding, pathogenic mutations were mapped to the modeled structure and subjected to 50ns molecular dynamics simulation. Our computational study suggests that four mutations are highly vulnerable to altering the structural conformation and stability of the PIGA protein, which illustrates its association with PNH and MCAHS2 phenotype.

Keywords: homology modeling, molecular dynamics simulation, missense mutations PNH, MCAHS2, PIGA

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Authors: Bandi Aruna

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The increase of multidrug-resistant pathogens and the restriction on the use of antibiotics due to its side effects have drawn attention to the search for possible alternatives. Bacteriocins are ribosomally synthesized antimicrobial peptides that are active against Gram-positive and Gram-negative bacteria. The bacteriocins from lactic acid bacteria represent an important application of these peptides as clinical drugs or as food biopreservatives. The present study describes the isolation of bacteriocin producing lactic acid bacteria (LAB) from fermented batter of green gram and bengal gram using Man, Rogosa and Sharpe (MRS) media. The bacteriocin produced by these organisms inhibited the growth of Staphylococcus aureus, Escherichia coli, Klebsiella species, Pseudomonas aeruginosa, The isolates G1, G2 were isolated from green gram; B1 and B2 were isolated from fermented bengal gram batter. G1 and G2 were identified as Lactobacillus casie and B1 and B2 were identified as Streptococcus species. Antimicrobial activity of the bacteriocin produced by these strains was studied by agar well diffusion method. Bacteriocins produced by the Lactobacillus casie and Streptococcus secies retained their antagonistic property at pH of 5 and pH of 7. Exposure of bacteriocin to UV light for 4 min showed antibacterial activity. The antagonistic property was observed even at 100°C demonstrating stability at higher temperatures of the bacteriocin. The bacteriocins were stable for a period of 15 days at 27°C. The bacteriocins of G1, G2, and B2 exhibited highest antagonistic activity at pH of 5 and B1 at pH of 7. Therefore, the bacteriocins of the isolates may find important application in controlling the food-borne pathogens.

Keywords: Keywords: Antibacterial activity, Lactic acid bacteria, Bacteriocin

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Authors: Ghebremedhin Tsegay, Weldu Tesfagaber, Yuanmao Zhu, Xijun He, Wan Wang, Zhenjiang Zhang, Encheng Sun, Jinya Zhang, Yuntao Guan, Fang Li, Renqiang Liu, Zhigao Bu, Dongming Zhao*

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African swine fever (ASF) is a highly infectious viral disease of pigs, resulting in significant economic loss worldwide. As there is no approved vaccines and treatments, the control of ASF entirely depends on early diagnosis and culling of infected pigs. Thus, highly specific and sensitive diagnostic assays are required for accurate and early diagnosis of ASF virus (ASFV). Currently, only a few recombinant proteins have been tested and validated for use as reagents in ASF diagnostic assays. The most promising ones for ASFV antibody detection were p72, p30, p54, and pp62. So far, three ELISA kits based on these recombinant proteins have been commercialized. Due to the complex nature of the virus and variety forms of the disease, robust serodiagnostic assays are still required. ASFV p22 protein, encoded by KP177R gene, is located in the inner membrane of viral particle and appeared transiently in the plasma membrane early after virus infection. The p22 protein interacts with numerous cellular proteins, involved in processes of phagocytosis and endocytosis through different cellular pathways. However, p22 does not seem to be involved in virus replication or swine pathogenicity. In this study, E.coli expressed recombinant p22 protein was used to generate a monoclonal antibody (mAb), and its potential use for the development of blocking ELISA (bELISA) was evaluated. A total of 806 pig serum samples were tested to evaluate the bELISA. Acording the ROC (Reciever operating chracteristic) analysis, 100% sensitivity and 98.10% of specificity was recorded when the PI cut-off value was set at 47%. The novel assay was able to detect the antibodies as early as 9 days post infection. Finaly, a highly sensitive, specific and rapid novel p22-mAb based bELISA assay was developed, and optimized for detection of antibodies against genotype I and II ASFVs. It is a promising candidate for an early and acurate detection of the antibodies and is highly expected to have a valuable role in the containment and prevention of ASF.

Keywords: ASFV, blocking ELISA, diagnosis, monoclonal antibodies, sensitivity, specificity

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Authors: Mohamed Radwan

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Introduction: Polycystic ovary syndrome is becoming very common among girls from ages 14 to 21 years old, alongside women in their childbearing period. A combination of menstrual irregularities, overweight or obesity, insulin resistance (type 2 diabetes), and symptoms of virilization are characteristics of these cases. Background: A multidisciplinary team should be involved in managing these cases and may consist of A gynecologist, endocrinologist, nutritionist, and psychologist. The role of nutritionists is crucial in these cases in weight reduction and nutritional management. It is mandatory to present some questions that may help in expanding further our understanding of the nutritional management of this syndrome, such as it is not an easy process to reduce the weight of PCO patients (2) we need to know what suitable nutritional plans for these cases are. Methodology: It is not successful to use one diet plan for all patients or a specific plan, but we need to give tailored plans for each patient. We will summarize the medical, nutritional therapy and weight management in polycystic ovary patients and highlight the best eating plan and dietary composition in the treatment of these women. We will also discuss the role of dieticians in treating polycystic ovary cases and overcoming the challenges these women face. Conclusion: All diet plans would decrease the weight of Polycystic ovary cases as low calory diet, low glycemic index diet, high protein diet, as well as Dash diet, but the most suitable diet plan to improve hormones and lead to spontaneous pregnancy is the Mediterranean diet.

Keywords: obesity, PCO mediteranian diet, dash diet, high protein diet

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