Search results for: isolation and protein malnutrition

Authors: Kang-Hyun Leem, Myung-Gyou Kim, Hye Kyung Kim

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Deer antler, traditionally used as a tonic and valuable drug in oriental medicine, has been considered to possess bone-strengthening activity. The upper section, mid section, and base of the antler has been known to exhibit different biological properties. Present study was performed to examine the effects of different parts of deer antler extract (DH) on osteogenic gene expressions in MG-63 cells and longitudinal bone growth in adolescent male rats. The expressions of osteogenic genes, collagen, alkaline phosphatase, osteocalcin, and osteopontin, were measured by quantitative real-time PCR. Longitudinal bone growth was measured in 3-week-old male Sprague-Dawley rats using fluorescence microscopy. To examine the effects on the growth plate metabolism, the total height of growth plate and bone morphogenetic protein-2 (BMP-2) were measured. Collagen and osteocalcin mRNA expressions were increased by all three parts of the DH treatment while osteopontin gene expression was not affected by any of the DH treatment. Alkaline phosphatase gene expression was increased by upper and mid part of DH while base part of DH fails to affect alkaline phosphatase gene expression. The upper and mid parts of the DH treatment enhanced longitudinal bone growth and total height of growth plate. The induction of BMP-2 protein expression in growth plate assessed by immunostaining was also promoted by upper and mid parts of the DH treatment. These results suggest that DH, especially upper and mid parts, stimulate osteogenic gene expressions and have the effect on bone growth in adolescent rats and might be used for the growth delayed adolescent and inherent growth failure patient.

Keywords: bone morphogenetic protein-2, deer antler, longitudinal bone growth, osteogenic genes

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Authors: Belinda Vega, Claudio Alvarez, Héctor Flores, Marcia Oliva, Katherine Alveal, Teresa Toro, María José Tapia, Fanny Guzmán

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With the increase in global temperatures and the decrease of oxygen (O2) concentration in the oceans, fish cultures are exposed to frequent fluctuations in dissolved O2 (DO) concentration that can cause chronic stress in the animals, altering the normal functioning of their immune system and making them vulnerable to infections, consequently increasing morbidity and mortality in the farms with economic losses. The mucosal organs (skin -and mucus-, gills, gut, and nasal mucosa) are the first line of defense of the fish against pathogens. Therefore, the objective of this study is to evaluate the effect of hypoxia on the antimicrobial activity of epidermal mucus from corvina drum (Cilus Gilberti), a native marine species with the potential for the diversification of aquaculture in Chile. To achieve this, the epidermal mucus of juveniles (~220g) kept under normoxia (7 mg/L DO) and hypoxia (2 mg/L DO) environmental conditions was collected after 6 weeks, as well as after 6 days of intraperitoneal inoculation with lipopolysaccharide from Vibrio anguillarum to induce an immune response in the fish. Total protein extracts of the mucus were used for bactericidal activity and lysozyme and peroxidase activity assays. Although the mucus from both experimental groups showed inhibitory effects on the bacterial growth of Vibrio anguillarum and Vibrio ordalli, this effect was more long-lasting in the normoxia group. We also observed a notable reduction in the presence of lysozyme in the mucus from fish exposed to hypoxia, with no differences in peroxidase content. Future proteomic studies of corvina mucus associated with the environmental conditions studied in this work will allow the isolation and identification of peptides with antimicrobial activity, those responsible for the results obtained. This will help establish strategies aimed at minimizing the impacts of hypoxia on the defense responses of corvina drum against potential pathogens. Funding: FONDECYT 3200440 and FONDECYT 1210056

Keywords: Cilus gilberti, mucus, antimicrobial activity, HYPOXIA

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Authors: Naveen Kumar B. T., Anuj Tyagi, Niraj Kumar Singh, Visanu Boonyawiwat, Shanthanagouda A. H., Orawan Boodde, Shankar K. M., Prakash Patil, Shubhkaramjeet Kaur

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Since 2009, Acute Hepatopancreatic Necrosis Disease (AHPND) outbreaks have increased rapidly, and these have led to the major economic losses to the global shrimp industry. In comparison to other treatments, passive immunity and monoclonal antibody (MAb) based farmer level kit have proved their importance in controlling and treating the diseases in the shrimp industry. In the present study, MAbs were produced against the recombinant PirB protein Vibrio parahaemolyticus strain causing AHPND. Briefly, Balb/C mice were immunized with rPirB at 15 days interval, and antibody titer was determined by ELISA. Spleen cells from mice showing high antibody titer were fused with SP2O myeloma cells for hybridoma production. Among 130 hybridomas, four showed high antibody titer and positive reactivity in an immunoblot assay. In Western blot assay, three out of four MAbs (4C4, 2C2 and 4G3) showed reactivity to rPirB protein. However, in the natural host, only Mab clone 4G3 show strong reactivity (with a strain of V. parahemolyticus causing EMS/AHPND). These clones also showed reactivity with less than 20 kDa proteins in AHPND free V. parahaemolyticus (Thailand stain). Further, on from MAb 4G3 clone, four panels of single cell MAbs clones (G3F5, G3B8, G3H2, and G3D6) were produced of which three showed strong positive reactivity to rPirB protein in the Western blot. These MAbs have potential for controlling and prevention of the AHPND through passive immunity and development of filed level rapid diagnostic kits.

Keywords: shrimp, economic loss, AHPND, MAb

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Authors: Maryam Farzaneh, Seyyedeh Nafiseh Hassani, Hossein Baharvand

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Objective: Chicken gonadal tissue has a two population such primordial germ cells (PGCs) and stromal cells (somatic cells). PGCs and embryonic germ cells (EGCs) that is a pluripotent type of PGCs in long-term culture are suitable sources for the production of chicken pluripotent stem cell lines, transgenic birds, vaccine and recombinant protein production. In general, the effect of growth factors such bFGF and mouse LIF on derivation of PGCs in vitro are important and in this study we could see the unique effect of small molecules such PD032 and SB43 as a chemical, in comparison to growth factors. Materials and Methods: After incubation of fertilized chicken egg up to 6 days and isolation of primary gonadal tissues and culture of mixed cells like PGCs and stromal cells. PGCs proliferate in the present of fetal calf serum (FCS) and small molecules and in another group bFGF, that these factors are important for PGCs culture and derivation. Somatic cells produce a multilayer feeder under the PGCs in primary culture and PGCs make a small cluster under these cells. Results: In present of small molecules and high volume of FCS (15%), the present of EGCs as a pluripotent stem cells were clear four weeks, that they had a positive immune-staining and periodic acid-Schiff staining (PAS), but in present of growth factors like bFGF without any chemicals, the present of PGCs were clear but after 7 until 10 days, there were disappear. Conclusion: Until now we have seen many researches about derivation and maintenance of chicken PGCs, in the hope of understanding the mechanisms that occur during germline development and production of a therapeutic product by transgenic birds. There are still many unknowns in this area and this project will try to have efficient conditions for identification of suitable culture medium for long-term culture of PGCs in vitro without serum and feeder cells.

Keywords: chicken gonadal primordial germ cells, pluripotent stem cells, growth factors, small molecules, transgenic birds

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Authors: Masataka Inada, Masanao Kinoshita, Nobuaki Matsumori

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It has been demonstrated that lipid molecules in biological membranes are responsible for the functionalization and structuration of membrane proteins. However, it is still unclear how the interaction of lipid molecules with membrane proteins is correlated with the function of the membrane proteins. Here we first developed an evaluation method for the interaction between membrane proteins and lipid molecules via surface plasmon resonance (SPR) analysis. Bacteriorhodopsin (bR), which was obtained by the culture of halobacteria, was used as a membrane protein. We prepared SPR sensor chips covered with self-assembled monolayer containing mercaptocarboxylic acids, and immobilized bR onto them. Then, we evaluated the interactions with various lipids that have different structures. As a result, the halobacterium-specific glycolipid S-TGA-1 was found to have much higher affinity with bRs than other lipids. This is probably due to not only hydrophobic and electrostatic interactions but also hydrogen bonds with sugar moieties in the glycolipid. Next, we analyzed the roles of the lipid in the structuration and functionalization of bR. CD analysis showed that S-TGA-1 could promote trimerization of bR monomers more efficiently than any other lipids. Flash photolysis further indicated that bR trimers formed by S-TGA-1 reproduced the photocyclic activity of bR in purple membrane, halobacterium-membrane. These results suggest that S-TGA-1 promotes trimerization of bR through strong interactions and consequently fulfills the bR’s function efficiently.

Keywords: membrane protein, lipid, interaction, bacteriorhodopsin, glycolipid

Procedia PDF Downloads 235

Authors: S. Dias, A. Tadeu, J. Antonio, F. Pedro, C. Serra

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The goal of the present work is to experimentally evaluate the feasibility of using vibration isolators made of expanded cork agglomerate. Even though this material, also known as insulation cork board (ICB), has mainly been studied for thermal and acoustic insulation purposes, it has strong potential for use in vibration isolation. However, the adequate design of expanded cork blocks vibration isolators will depend on several factors, such as excitation frequency, static load conditions and intrinsic dynamic behavior of the material. In this study, transmissibility tests for different static and dynamic loading conditions were performed in order to characterize the material. Since the material’s physical properties can influence the vibro-isolation performance of the blocks (in terms of density and thickness), this study covered four mass density ranges and four block thicknesses. A total of 72 expanded cork agglomerate specimens were tested. The test apparatus comprises a vibration exciter connected to an excitation mass that holds the test specimen. The test specimens under characterization were loaded successively with steel plates in order to obtain results for different masses. An accelerometer was placed at the top of these masses and at the base of the excitation mass. The test was performed for a defined frequency range, and the amplitude registered by the accelerometers was recorded in time domain. For each of the signals (signal 1- vibration of the excitation mass, signal 2- vibration of the loading mass) a fast Fourier transform (FFT) was applied in order to obtain the frequency domain response. For each of the frequency domain signals, the maximum amplitude reached was registered. The ratio between the amplitude (acceleration) of signal 2 and the amplitude of signal 1, allows the calculation of the transmissibility for each frequency. Repeating this procedure allowed us to plot a transmissibility curve for a certain frequency range. A number of transmissibility experiments were performed to assess the influence of changing the mass density and thickness of the expanded cork blocks and the experimental conditions (static load and frequency of excitation). The experimental transmissibility tests performed in this study showed that expanded cork agglomerate blocks are a good option for mitigating vibrations. It was concluded that specimens with lower mass density and larger thickness lead to better performance, with higher vibration isolation and a larger range of isolated frequencies. In conclusion, the study of the performance of expanded cork agglomerate blocks presented herein will allow for a more efficient application of expanded cork vibration isolators. This is particularly relevant since this material is a more sustainable alternative to other commonly used non-environmentally friendly products, such as rubber.

Keywords: expanded cork agglomerate, insulation cork board, transmissibility tests, sustainable materials, vibration isolators

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Authors: Ying Sun, Biao Cheng, Qing Lu, Xuefei Tao, Xiaoyu Lai, Cheng Guo, Dan Wang

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Fibrinogen-like protein 2 (FGL2) has recently been identified to play an important role in inflammatory diseases such as atherosclerosis through a thrombin-dependent manner. Here, a murine monoclonal antibody was raised against the critical residue Ser(89) of FGL2, and the effects of the anti-FGL2 mAb (SPF89) were analyzed in human umbilical vein endothelial cells (HUVECs) and THP-1 cells. Firstly, it was proved that SPF89, which belongs to the IgG1 subtype with a KD value of 44.5 pM, could specifically show the expression levels of protein FGL2 in different cell lines of known target gene status. The lipopolysaccharide (LPS)-mediated endothelial cell proliferation was significantly inhibited with a decline of phosphorylation nuclear factor-κB (NF-κB) in a dose-dependent manner after SPF89 treatment. Furthermore, SPF89 reduced LPS-induced expression of adhesion molecules and inflammatory cytokines such as vascular cell adhesion molecule-1, tumor necrosis factor-α, Matrix metalloproteinase MMP-2, Integrin αvβ3, and interleukin-6 in HUVECs. In macrophage-like THP-1 cells, SPF89 effectively inhibited LPS and low-density lipoprotein-induced foam cell formation. However, these anti-inflammatory and anti-atherosclerotic effects of anti-FGL2 mAb in HUVECs and THP-1 cells were significantly reduced after treatment with an NF-κB inhibitor PDTC. All the above suggest, by efficiently inhibiting LPS-induced pro-inflammatory effects in vascular endothelial cells by attenuating NF-κB dependent pathway, the new anti-FGL2 mAb SPF89 could to be a potential therapeutic candidate for protecting the vascular endothelium against inflammatory diseases such as atherosclerosis. This work was supported by the Program of Sichuan Science and Technology Department (2017FZ0069) and Collaborative Innovation Program of Sichuan for Elderly Care and Health(YLZBZ1511).

Keywords: monoclonal antibody, fibrinogen like protein 2, inflammation, endothelial cells

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Authors: Akshita Bhardwaj

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Keratinase is an enzyme obtained from extracellular sources that is involved in biodegradation of keratin. It is a member of a group of proteases that can break down keratin into amino acids. Keratinases are produced only in the presence of substrate that contain keratin. It attacked the disulfide bond of substrate and involve in keratin degradation. Human hair, feathers, animal hard tissues, horns, claws, and hooves all contain keratin.. It exists in two form alpha keratin (found in soft tissues) and beta keratin (found in hard tissue). By taking part in the degradation of keratin, keratinases derived from microbial sources, often referred to as microbial keratinases, are important in the process of turning wastes containing keratin into products with added value. Chicken feathers contain high level of keratin protein content than other sources and became a suitable protein source. Keratinase production occurs at near alkaline pH and thermophilic temperatures. The bioprocessing of keratinous waste benefits greatly from the use of keratinases. Additionally, it lessens the issue caused by poultry excrement. The use of feather meal, along with keratinase, improves the digestion of proteins and amino acids.

Keywords: mili litre (ml), micro litre (Ul), TCA - trichloroacetic acid, OD - optical density

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Authors: Joo-min Kim, Gi-wook Shin, Tae-ho Chung, Chul Park, Seong-hyun Kim, Namjung Kim

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Mealworm (Tenebrio molitor) was evaluated to investigate the effect of partial or total replacement of fish meal in diets for olive flounder, Paralichthys olivaceus. Experimental groups of fish with average initial body weight (287.5 ± 7.24 g) were fed each with 4 isonitrogeneous (52% crude protein) diets formulated to include 0, 7, 17 and 27% (diets 1 to 4, respectively) of fish meal substituted with mealworm. After six weeks of feeding trials, fish fed with diet 3 revealed the highest values for live weight gain(42.10), specific growth rates (0.445 ± 0.089) as well as better feed conversion ratio (12.08) compared to the other group with statistically significant manner (p<0.05). Hepatosomatic index was showed no significant difference in diet 3 compared to the control group. An increase in weight gain and other growth associated parameters was observed in diet 3. These results clearly indicate that 17% of fish meal protein in bastard halibut diet can be replaced by mealworm not only without any adverse effect but also the effect of promoting growth performance.

Keywords: mealworm, olive flounder, Paralichthys olivaceus, Tenebrio molitor

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Authors: Aqeel Ahmed Shad, Tanveer Ahmad, Muhammad Farooq Iqbal, Muhammad Javaid Asad

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The supplemental effects of novel protease produced from Bacillus subtilis K-5 and beta-mannanase were evaluated on growth performance, carcass characteristics, nutrients digestibility, blood profile and intestinal morphometry of broilers fed guar meal (Cyamopsis tetragonoloba) based diets with reduced Crude Protein (CP), Essential Amino Acids (EAAs), and Metabolizable energy (ME) contents. One-day old Ross 308 broiler chicks (n=360) were randomly allotted to thirty six experimental units in a way that each of the nine dietary treatments received four replicates with ten birds per replicate. A control diet without guar meal (0GM) was formulated with standard nutrient specifications of Ross 308 for the starter and finisher phases. Two negative control diets, one with 5% (5GM) and second with 10% (10GM) guar meal, were formulated with reduction of 5% CP, 5% EAAs and 80 Kcal/kg ME. These three basal diets (no enzyme) were supplemented with novel protease enzyme (PROT) and commercial beta-mannanase (Beta-M) enzyme. The birds were reared up to 35d of age. The data on weekly body weight gain (BWG) and feed intake were recorded to compute feed:gain for the starter (0-21d) and finisher (22-35d) phases. At the end of 35d of experimental period, four birds per experimental unit were randomly selected for blood samples collection and later slaughtered for ileal digesta, intestinal tract and carcass trait sampling. The data on overall performance (1-35d) indicated improved (P<0.05) BWG and feed:gain in birds supplemented with PROT (1.41% and 1.67) and Beta-M (2.79% and 1.64) than non-supplemented groups. Improved (P<0.05) carcass yield, breast meat yield and thigh meat yield were noted with the supplementation of Beta-M. However, non-significant (P>0.05) effect on carcass traits was noted in broiler fed guar meal based PROT supplemented diets. Crude protein digestibility, nitrogen retention (Nret) and apparent digestibility coefficient for nitrogen (ADCN) were improved (P<0.05) only with PROT. The improvement in apparent metabolizable energy (AME) and apparent metabolizable energy corrected for nitrogen (AMEn) was noted (P<0.05) with both supplemented enzymes. However, no effect (P>0.05) of enzyme addition was noted on blood glucose, total protein and cholesterol. Improved villus height of duodenum, jejunum and ileum was noted (P<0.05) with the addition of both enzymes. The EAAs digestibility was improved (P<0.05) only with PROT. In conclusion, beta-mannanase and protease supplementation better improved the overall bird performance in low nutrient profile guar meal based diets than non-supplemented diets.

Keywords: novel protease, guar meal, broilers, low protein diets, low metabolizable energy diets, nutrients digestibility

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Authors: Andrea Tonoli, Nicola Amati, Maria Cavatorta, Reza Mirsanei, Behzad Mozaffari, Hamed Ahani, Akbar Karamihafshejani, Mohammad Ghazivakili, Mohammad Abuabiah

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The vibrations transmitted to the drivers and passengers through vehicle seat seriously effect on the level of their attention, fatigue and physical health and reduce the comfort and efficiency of the occupants. Recently, some researchers have focused on vibrations at low excitation frequency(0.5-5 Hz) which are considered to be the main risk factors for lumbar part of the backbone but they were not applicable to A and B-segment cars regarding to the size and weight. A semi-active system with two symmetric negative stiffness structures (NSS) in parallel to a positive stiffness structure and actuators has been proposed to attenuate low frequency excitation and makes system flexible regarding to different weight of passengers which is applicable for A and B-Segment cars. Here, the 3 degree of freedom system is considered, dynamic equation clearly is presented, then simulated in MATLAB in order to analysis of performance of the system. The design procedure is derived so that the resonance peak of frequency–response curve shift to the left, the isolating range is increased and especially, the peak of the frequency–response curve is minimized. According to ISO standard different class of road profile as an input is applied to the system to evaluate the performance of the system. To evaluate comfort issues, we extract the RMS value of the vertical acceleration acting on the passenger's body. Then apply the band-pass filter, which takes into account the human sensitivity to acceleration. According to ISO, this weighted acceleration is lower than 0.315 m/s^2, so the ride is considered as comfortable.

Keywords: low frequency excitation, negative stiffness, seat vehicle, vibration isolation

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Authors: Mona G. Alharbi

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A new family of methionine-sulfoxide reductase (Msr) was recently discovered and was named free methionine sulfoxide reductase (fRMsr). This family includes enzymes with a reductase activity toward the free R isomer of a methionine sulfoxide substrate. The fRMsrs have a GAF domain topology, a domain, which was previously identified as having in some cases a cyclic nucleotide phosphodiesterase activity. The classification of fRMsrs as GAF domains revealed a new function can be added to the GAF domain family. Interestingly the four members identified in the fRMsr family share the GAF domain structure and the presence of three conserved cysteines in the active site with free R methionine sulfoxide substrate specificity. This thesis presents the crystal structures of reduced, free Met-SO substrate-bound and MES-bound forms of a new fRMsr from Burkholderia pseudomallei (BPSL2418). BPSL2418 was cloned, overexpressed and purified to enable protein crystallization. The crystallization trials for reduced, Met-SO-bound and MES-bound forms of BPSL2418 were prepared and reasonable crystals of each form were produced. The crystal structures of BPSL2418MES, BPSL2418Met-SO and BPSL2418Reduced were solved at 1.18, 1.4 and 2.0Å, respectively by molecular replacement. The BPSL2418MES crystal belongs to space group P 21 21 21 while BPSL2418Met-SO and BPSL2418Reduced crystals belong to space group P 1 21 1. All three forms share the GAF domain structure of six antiparallel β-strands and four α-helices with connecting loops. The antiparallel β-strands (β1, β2, β5 and β6) are located in the center of the BPSL2418 structure flanked on one side by a three α-helices (α1, α2 and α4) and on the other side by a (loop1, β3, loop2, α3, β4 loop4) unit where loop4 forms a capping flap and covers the active site. The structural comparison of the three forms of BPSL2418 indicates that the catalytically important cysteine is CYS109, where the resolving cysteine is CYS75, which forms a disulfide bond with CYS109. They also suggest that the third conserved cysteine in the active site, CYS85, which is located in α3, is a non-essential cysteine for the catalytic function but it may play a role in the binding of the substrate. The structural comparison of the three forms reveals that conformational changes appear in the active site particularly involving loop4 and CYS109 during catalysis. The 3D structure of BPSL2418 shows strong structure similarity to fRMsrs enzymes, which further suggests that BPSL2418 acts as a free Met-R-SO reductase and shares the catalytic mechanism of fRMsr family.

Keywords: Burkholderia pseudomallei, GAF domain protein, methionine sulfoxide reductase, protein crystallization

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Authors: Tesfaye Walle Mekonnen

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Maize is one of the most important staple food crops for most low-income households in the Sub-Saharan (SSA). Combined heat and drought stress is the major production threats that reduce the yield potential of biofortified maize and restrain various macro and micronutrient deficiencies highly prevalent in low-income people who rely solely on maize-based diets, SSA. This problem can be alleviated by crossing the biofortified inbred lines with different nutritional attributes, Fe, Zn, Protein, and Provitamin A, and developing agronomically superior and stable multi-nutrient maize of various genetic backgrounds. This aimed to understand the correlation between biofortified inbred lines per se and hybrid performance under combined heat and drought stress conditions (CSC). The experiment was conducted at CIMMYT, Zimbabwe, using α-lattice design with three replications. The hybrid effect was highly significant for zein fractions (α-, β-, γ- and δ-zein) zinc, (Zn), and iron (Fe) provitamin A, phytic acid, and grain yield. Under CSC, Fe, Zn concentration, provitamin A in grain and grain yield of hybrids were significantly decreased, however, the zein fraction content and phytic acid content increases in grain were increased under CSC. The phenotypic correlation between grain yield with Zn, Fe concentration, and Provitamin A in grain was strongly positive and higher under CSC than in well-watered conditions. The present investigation confirmed that under CSC, Fe, and Zn-enhanced hybrids could be forecasted to a certain scope based on the performance of and scientifically selected for desirable grain yield and related traits with CSC tolerance during hybrid development programs. In conclusion, the development of high-yielding and micronutrient-dense maize variety is possible under CSC, which could reduce the highly prevalent micronutrient in SSA.

Keywords: drought, Fe, heat, maize, protein, zein fractions, Zn

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Authors: Irshad Ali Khan, Jian-Ping Lu, Xiao-Hong Liu, Fu-Cheng Lin

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This study reports the functional analysis of a gene MoNUC1 in M. oryzae, which is homologous to the Saccharomyces cerevisiae NUC1 encoding a mitochondrial nuclease protein. The MoNUC1 having a gene locus MGG_05324 is 1002-bp in length and encodes an identical protein of 333 amino acids. We disrupted the gene through gene disruption strategy and isolated two mutants confirmed by southern blotting. The deleted mutants were then used for phenotypic studies and their phenotypes were compared to those of the Guy-11 strain. The mutants were first grown on CM medium to find the effect of MoNUC1 gene disruption on colony growth and the mutants were found to show normal culture colony growth similar to that of the Guy-11 strain. Conidial germination and appressorial formation were also similar in both the mutants and Guy-11 strains showing that this gene plays no significant role in these phenotypes. For pathogenicity, the mutants and Guy-11 mycelium blocks were inoculated on blast susceptible barley seedlings and it was found that both the strains exhibited full pathogenicity showing coalesced and necrotic blast lesions suggesting that this gene is not involved in pathogenicity. Mating of the mutants with 2539 strain formed numerous perithecia showing that MoNUC1 is not essential for sexual reproduction in M. oryzae. However, the mutants were found to form reduced conidia (1.06±8.03B and 1.08±9.80B) than those of the Guy-11 strain (1.46±10.61A) and we conclude that this protein is not required for the blast fungus to cause pathogenicity but plays significant role in conidiation. Proteins of signal transduction pathways that could be disrupted/ intervened genetically or chemically could lead to antifungal products of important fungal cereal diseases and reduce rice yield losses. Tipping the balance toward understanding the whole of pathogenesis, rather than simply conidiation will take some time, but clearly presents the most exciting challenge of all.

Keywords: appressorium formation, conidiation, NUC1, Magnaporthe oryzae, pathogenicity

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Authors: Fidelis E. Uwumiro, Chimaobi O. Nwevo, Favour O. Osemwota, Victory O. Okpujie, Emeka S. Obi, Omamuyovbi F. Nwoagbe, Ejiroghene Tejere, Joycelyn Adjei-Mensah, Christopher N. Ekeh, Charles T. Ogbodo

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Several risk factors associated with cardiovascular events have been identified as specific to Chronic Kidney Disease (CKD). This study endeavors to validate these CKD-specific risk factors using up-to-date national-level data, thereby highlighting the crucial significance of confirming the validity and generalizability of findings obtained from previous studies conducted on smaller patient populations. The study utilized the nationwide inpatient sample database to identify adult hospitalizations for CKD from 2016 to 2020, employing validated ICD-10-CM/PCS codes. A comprehensive literature review was conducted to identify both traditional and CKD-specific risk factors associated with cardiovascular events. Risk factors and cardiovascular events were defined using a combination of ICD-10-CM/PCS codes and statistical commands. Only risk factors with specific ICD-10 codes and hospitalizations with complete data were included in the study. Cardiovascular events of interest included cardiac arrhythmias, sudden cardiac death, acute heart failure, and acute coronary syndromes. Univariate and multivariate regression models were employed to evaluate the association between chronic kidney disease-specific risk factors and cardiovascular events while adjusting for the impact of traditional CV risk factors such as old age, hypertension, diabetes, hypercholesterolemia, inactivity, and smoking. A total of 690,375 hospitalizations for CKD were included in the analysis. The study population was predominantly male (375,564, 54.4%) and primarily received care at urban teaching hospitals (512,258, 74.2%). The mean age of the study population was 61 years (SD 0.1), and 86.7% (598,555) had a CCI of 3 or more. At least one traditional risk factor for CV events was present in 84.1% of all hospitalizations (580,605), while 65.4% (451,505) included at least one CKD-specific risk factor for CV events. The incidence of CV events in the study was as follows: acute coronary syndromes (41,422; 6%), sudden cardiac death (13,807; 2%), heart failure (404,560; 58.6%), and cardiac arrhythmias (124,267; 18%). 91.7% (113,912) of all cardiac arrhythmias were atrial fibrillations. Significant odds of cardiovascular events on multivariate analyses included: malnutrition (aOR: 1.09; 95% CI: 1.06–1.13; p<0.001), post-dialytic hypotension (aOR: 1.34; 95% CI: 1.26–1.42; p<0.001), thrombophilia (aOR: 1.46; 95% CI: 1.29–1.65; p<0.001), sleep disorder (aOR: 1.17; 95% CI: 1.09–1.25; p<0.001), and post-renal transplant immunosuppressive therapy (aOR: 1.39; 95% CI: 1.26–1.53; p<0.001). The study validated malnutrition, post-dialytic hypotension, thrombophilia, sleep disorders, and post-renal transplant immunosuppressive therapy, highlighting their association with increased risk for cardiovascular events in CKD patients. No significant association was observed between uremic syndrome, hyperhomocysteinemia, hyperuricemia, hypertriglyceridemia, leptin levels, carnitine deficiency, anemia, and the odds of experiencing cardiovascular events.

Keywords: cardiovascular events, cardiovascular risk factors in CKD, chronic kidney disease, nationwide inpatient sample

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Authors: Simona Dostalova, Pavel Kopel, Marketa Vaculovicova, Vojtech Adam, Rene Kizek

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Protein apoferritin seems to be a very promising structure for use as a nanocarrier. It is prepared from intracellular ferritin protein naturally found in most organisms. The role of ferritin proteins is to store and transport ferrous ions. Apoferritin is a hollow protein cage without ferrous ions that can be prepared from ferritin by reduction with thioglycolic acid or dithionite. The structure of apoferritin is composed of 24 protein subunits, creating a sphere with 12 nm in diameter. The inner cavity has a diameter of 8 nm. The drug encapsulation process is based on the response of apoferritin structure to the pH changes of surrounding solution. In low pH, apoferritin is disassembled into individual subunits and its structure is “opened”. It can then be mixed with any desired cytotoxic drug and after adjustment of pH back to neutral the subunits are reconnected again and the drug is encapsulated within the apoferritin particles. Excess drug molecules can be removed by dialysis. The receptors for apoferritin, SCARA5 and TfR1 can be found in the membrane of both healthy and cancer cells. To enhance the specific targeting of apoferritin nanocarrier, it is possible to modify its surface with targeting moieties, such as antibodies. To ensure sterically correct complex, we used a a peptide linker based on a protein G with N-terminus affinity towards Fc region of antibodies. To connect the peptide to the surface of apoferritin, the C-terminus of peptide was made of cysteine with affinity to gold. The surface of apoferritin with encapsulated doxorubicin (ApoDox) was coated either with gold nanoparticles (ApoDox-Nano) or gold (III) chloride hydrate reduced with sodium borohydride (ApoDox-HAu). The applied amount of gold in form of gold (III) chloride hydrate was 10 times higher than in the case of gold nanoparticles. However, after removal of the excess unbound ions by electrophoretic separation, the concentration of gold on the surface of apoferritin was only 6 times higher for ApoDox-HAu in comparison with ApoDox-Nano. Moreover, the reduction with sodium borohydride caused a loss of doxorubicin fluorescent properties (excitation maximum at 480 nm with emission maximum at 600 nm) and thus its biological activity. Fluorescent properties of ApoDox-Nano were similar to the unmodified ApoDox, therefore it was more suited for the intended use. To evaluate the specificity of apoferritin modified with antibodies, we used ELISA-like method with the surface of microtitration plate wells coated by the antigen (goat anti-human IgG antibodies). To these wells, we applied ApoDox without targeting antibodies and ApoDox-Nano modified with targeting antibodies (human IgG antibodies). The amount of unmodified ApoDox on antigen after incubation and subsequent rinsing with water was 5 times lower than in the case of ApoDox-Nano modified with targeting antibodies. The modification of non-gold ApoDox with antibodies caused no change in its targeting properties. It can therefore be concluded that the demonstrated procedure allows us to create nanocarrier with enhanced targeting properties, suitable for nanomedicine.

Keywords: apoferritin, doxorubicin, nanocarrier, targeting antibodies

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Authors: Mirnawati, Gita Ciptaan, Ferawati

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Background and Objective: Palm kernel cake (PKC) is a waste of the palm oil industry. Indonesia, as the largest palm oil producer in the world, produced 45-46% palm kernel cake. Palm kernel cake can potentially be used as animal ration but its utilization for poultry is limited. Thus, fermentation process was done in order to increase the utilization PKC in poultry ration. An experiment was conducted to study the effect between Inoculum Doses with Bacillus subtilis and fermentation time to improve the quality and nutrient content of fermented Palm Kernel Cake. Material and Methods: 1) Palm kernel cake derived from Palm Kernel Processing Manufacture of Andalas Agro Industry in Pasaman, West Sumatra. 2) Bacillus subtilis obtained from The Research Center of Applied Chemistry LIPI, Bogor. 3) Preparations nutrient agar medium (NA) produced by Difoo - Becton Dickinson. 4) Rice bran 5) Aquades and mineral standard. The experiment used completely randomize design (CRD) with 3 x 3 factorial and 3 replications. The first factors were three doses of inoculum Bacillus subtilis: (3%), (5%), and (7%). The second factor was fermentation time: (1) 2 day, (2) 4 day, and (3) 6 day. The parameters were crude protein, crude fiber, nitrogen retention, and crude fiber digestibility of fermented palm kernel cake (FPKC). Results: The result of the study showed that there was significant interaction (P<0.01) between factor A and factor B and each factor A and B also showed significant effect (P<0.01) on crude protein, crude fiber, nitrogen retention, and crude fiber digestibility. Conclusion: From this study, it can be concluded that fermented PKC with 7% doses of Bacillus subtilis and 6 days fermentation time provides the best result as seen from 24.65% crude protein, 17.35% crude fiber, 68.47% nitrogen retention, 53.25% crude fiber digestibility of fermented palm kernel cake (FPKC).

Keywords: fermentation, Bacillus Subtilis, inoculum, palm kernel cake, quality, nutrient

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Authors: Brian W. Mandikiana

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Homestead food production has the potential to alleviate hunger, improve health and nutrition for children and adults. This article examines the relationship between self-reported hunger and homestead food production using the ordered probit model. A sample of households participating in homestead food production was drawn from the first wave of the South African National Income Dynamics Survey, a nationally representative cross-section. The sample selection problem was corrected using an ordered probit model with sample selection approach. The findings show that homestead food production exerts a positive and significant impact on children and adults’ ability to cope with hunger and malnutrition. Yet, on the contrary, potential gains of homestead food production are threatened by shocks such as crop failure.

Keywords: agriculture, hunger, nutrition, sample selection

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Authors: Şeyma Özçirak Ergün, Ergün Şakalar, Emrah Yalazi̇, Nebahat Şahi̇n

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Food irradiation is a usage of exposing food to ionising radiation (IR) such as gamma rays. IR has been used to decrease the number of harmful microorganisms in the food such as spices. Excessive usage of IR can cause damage to both food and people who consuming food. And also it causes to damages on food DNA. Generally, IR detection techniques were utilized in literature for spices are Electron Spin Resonance (ESR), Thermos Luminescence (TL). Storage creates negative effect on IR detection method then analyses of samples have been performed without storage in general. In the experimental part, red pepper (Capsicum annuum) and mint (Mentha piperita) as spices were exposed to 0, 0.272, 0.497, 1.06, 3.64, 8.82, and 17.42 kGy ionize radiation. ESR was applied to samples irradiated. DNA isolation from irradiated samples was performed using GIDAGEN Multi Fast DNA isolation kit. The DNA concentration was measured using a microplate reader spectrophotometer (Infinite® 200 PRO-Life Science–Tecan). The concentration of each DNA was adjusted to 50 ng/µL. Genomic DNA was imaged by UV transilluminator (Gel Doc XR System, Bio-Rad) for the estimation of genomic DNA bp-fragment size after IR. Thus, agarose gel profiles of irradiated spices were obtained to determine the change of band profiles. Besides, samples were examined at three different time periods (0, 3, 6 months storage) to show the feasibility of developed method. Results of gel electrophoresis showed especially degradation of DNA of irradiated samples. In conclusion, this study with gel electrophoresis can be used as a basis for the identification of the dose of irradiation by looking at degradation profiles at specific amounts of irradiation. Agarose gel results of irradiated samples were confirmed with ESR analysis. This method can be applied widely to not only food products but also all biological materials containing DNA to predict radiation-induced damage of DNA.

Keywords: DNA, electrophoresis, gel electrophoresis, ionizeradiation

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Authors: Sarmiza Stanca, Wolfgang Fritzsche, Christoph Krafft, Jürgen Popp

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Independent of the cell type a thin layer of a few nanometers thickness surrounds the cell interior as the cellular membrane. The transport of ions and molecules through the membrane is achieved in a very precise way by pores. Understanding the process of opening and closing the pores due to an electrochemical gradient across the membrane requires knowledge of the pore constitutive proteins. Recent reports prove the access to the molecular level of the cellular membrane by atomic force microscopy (AFM). This technique also permits an electrochemical study in the immediate vicinity of the tip. Specific molecules can be electrochemically localized in the natural cellular membrane. Our work aims to recognize the protein domains of the pores using an AFM probe as a miniaturized amperometric sensor, and to follow the protein behavior while changing the applied potential. The intensity of the current produced between the surface and the AFM probe is amplified and detected simultaneously with the surface imaging. The AFM probe plays the role of the working electrode and the substrate, a conductive glass on which the cells are grown, represent the counter electrode. For a better control of the electric potential on the probe, a third electrode Ag/AgCl wire is mounted in the circuit as a reference electrode. The working potential is applied between the electrodes with a programmable source and the current intensity in the circuit is recorded with a multimeter. The applied potential considers the overpotential at the electrode surface and the potential drop due to the current flow through the system. The reported method permits a high resolved electrochemical study of the protein domains on the living cell membrane. The amperometric map identifies areas of different current intensities on the pore depending on the applied potential. The reproducibility of this method is limited by the tip shape, the uncontrollable capacitance, which occurs at the apex and a potential local charge separation.

Keywords: AFM, sensor, membrane, pores, proteins

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Authors: H. Mansoori Yarahmadi, A. Aghazadeh, K. Nazeradl

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This study was done to evaluate the effects of feeding canola seed for enrichment of UFA and milk performance of early lactation dairy cows. Twelve multi parous Holstein cows (635.3±18 kg BW and 36±9 DIM) were assigned to 1 of 3 treatments: 1- Control (CON) without canola seed, 2- 7.5% raw canola seed (CUT), and 3- 7.5% Heat-treated canola seed (CHT) of the total ration. Diets contained same crude protein, but varied in net energy. Diets were composed by basis of corn silage and alfalfa. Cows were milked twice daily for 4 wk. The inclusion of canola seed did not alter DM intake, weight gain, or body condition score of cows. Milk fat from CHT cows had greater proportions of UFA and MUFA (P < 0.05). Feeding CUT increased PUFA without significant difference. Milk fat from CHT had a greater proportion of C18 UFA and tended to have a higher proportion of other UFA. FCM milk yields, milk fat and protein percentages and total yield of these components were similar between treatments. Milk urea nitrogen was lower in cows fed CON and CHT. Feeding canola seed to lactating dairy cows resulted in milk fat with higher proportions of healthful fatty acids without adverse affecting milk yield or milk composition.

Keywords: canola seed, fatty acid, dairy cow, milk

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Authors: Silvia Gastaldello, Maria Grillo, Luca Tassoni, Claudio Maran, Stefano Balbo

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Antioxidants are nowadays attractive not only for the countless benefits to the human and animal health, but also for the perspective of use as food preservative instead of synthetic chemical molecules. In this study, the radical scavenging activity of six protein extracts from pulse and oleaginous seeds was evaluated. The selected matrices are Pisum sativum (yellow pea from two different origins), Carthamus tinctorius (safflower), Helianthus annuus (sunflower), Lupinus luteus cv Mister (lupin) and Glycine max (soybean), since they are economically interesting for both human and animal nutrition. The seeds were grinded and proteins extracted from 20mg powder with a specific vegetal-extraction kit. Proteins have been quantified through Bradford protocol and scavenging activity was revealed using DPPH assay, based on radical DPPH (2,2-diphenyl-1-picrylhydrazyl) absorbance decrease in the presence of antioxidants molecules. Different concentrations of the protein extract (1, 5, 10, 50, 100, 500 µg/ml) were mixed with DPPH solution (DPPH 0,004% in ethanol 70% v/v). Ascorbic acid was used as a scavenging activity standard reference, at the same six concentrations of protein extracts, while DPPH solution was used as control. Samples and standard were prepared in triplicate and incubated for 30 minutes in dark at room temperature, the absorbance was read at 517nm (ABS30). Average and standard deviation of absorbance values were calculated for each concentration of samples and standard. Statistical analysis using t-students and p-value were performed to assess the statistical significance of the scavenging activity difference between the samples (or standard) and control (ABSctrl). The percentage of antioxidant activity has been calculated using the formula [(ABSctrl-ABS30)/ABSctrl]*100. The obtained results demonstrate that all matrices showed antioxidant activity. Ascorbic acid, used as standard, exhibits a 96% scavenging activity at the concentration of 500 µg/ml. At the same conditions, sunflower, safflower and yellow peas revealed the highest antioxidant performance among the matrices analyzed, with an activity of 74%, 68% and 70% respectively (p < 0.005). Although lupin and soybean exhibit a lower antioxidant activity compared to the other matrices, they showed a percentage of 46 and 36 respectively. All these data suggest the possibility to use undervalued edible matrices as antioxidants source. However, further studies are necessary to investigate a possible synergic effect of several matrices as well as the impact of industrial processes for a large-scale approach.

Keywords: antioxidants, DPPH assay, natural matrices, vegetal proteins

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Authors: Benjaporn Buranrat, Nootchanat Mairuae

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Cratoxylum formosum (Jack) Dyer (CF) has been used for the traditional medicines in South East Asian and Thailand. Normally, northeast Thai vegetables have proven cytotoxic to many cancer cells. Therefore, the present study aims to explore the molecular mechanisms underlying CF-induced cancer cell death and apoptosis on breast and liver cancer cells. The cytotoxicity and antiproliferative effects of CF on the human breast MCF-7 and liver HepG2 cancer cell lines were evaluated using sulforhodamine B assay and colony formation assay. Cell migration assay was measured using wound healing assay. The apoptosis induction mechanisms were investigated through reactive oxygen species formation, caspase 3 activity, and JC-1 activity. Gene expression by real-time PCR and apoptosis related protein levels by Western blot analysis. CF induced MCF-7 and HepG2 cell death by time- and dose-dependent manner. Furthermore, CF had the greater cytotoxic potency on MCF-7 more than HepG2 cells with IC50 values of 85.70+4.52 μM and 219.03±9.96 μM respectively, at 24 h. Treatment with CF also caused a dose-dependent decrease in colony forming ability and cell migration, especially on MCF-7 cells. CF induced ROS formation, increased caspase 3 activities, and decreased the mitochondrial membrane potential, and causing apoptotic body production and DNA fragmentation. CF significantly decreased expression of the cell cycle regulatory protein RAC1 and downstream proteins, cdk6. Additionally, CF enhanced p21 and reduced cyclin D1 protein levels. CF leaf extract induced cell death, apoptosis, antimigration in both of MCF-7 and HepG2 cells. CF could be useful for developing to anticancer drug candidate for breast and liver cancer therapy.

Keywords: cratoxylum formosum (jack) dyer, breast cancer, liver cancer, cell death

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Authors: M. L. Mangena, N. Mokoena, K. Rashamuse, M. G. Tlou

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Tertiary alcohol ester (TAE) hydrolases are a group of esterases (EC 3.1.1.-) that catalyze the kinetic resolution of TAEs and as a result, they are sought-after for the production of optically pure tertiary alcohols (TAs) which are useful as building blocks for number biologically active compounds. What sets these enzymes apart is, the presence of a GGG(A)X-motif in the active site which appears to be the main reason behind their activity towards the sterically demanding TAEs. The genome of Pseudomonas syringae pv. maculicola (Psm) comprises a multitude of genes that encode esterases. We therefore, hypothesize that some of these genes encode TAE hydrolases. In this study, Psm was screened for TAE hydrolase activity using the linalyl acetate (LA) plate assay and a positive reaction was observed. As a result, the genome of Psm was screened for esterases with a GGG(A)X-motif using the motif search tool and two potential TAE hydrolase genes (PsmEST1 and 2, 1100 and 1000bp, respectively) were identified, PsmEST1 was amplified by PCR and the gene sequenced for confirmation. Analysis of the sequence data with the SingnalP 4.1 server revealed that the protein comprises a signal peptide (22 amino acid residues) on the N-terminus. Primers specific for the gene encoding the mature protein (without the signal peptide) were designed such that they contain NdeI and XhoI restriction sites for directional cloning of the PCR products into pET28a. The gene was expressed in E. coli JM109 (DE3) and the clones screened for TAE hydrolase activity using the LA plate assay. A positive clone was selected, overexpressed and the protein purified using nickel affinity chromatography. The activity of the esterase towards LA was confirmed using thin layer chromatography.

Keywords: hydrolases, tertiary alcohol esters, tertiary alcohols, screening, Pseudomonas syringae pv., maculicola genome, esterase activity, linalyl acetate

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Authors: Pratibha Goyal, Nupur Mathur, Anuradha Singh

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Antibiotic resistance is the worldwide threat to human health in this century. Excessive use of antibiotic after their discovery in 1940 makes certain bacteria to become resistant against antibiotics. Most common antibiotic-resistant bacteria include Staphylococcus aureus, Salmonella typhi, E.coli, Klebsiella pneumonia, and Streptococcus pneumonia. Among all Staphylococcus resistant strain called Methicillin-resistant Staphylococcus aureus (MRSA) is responsible for several lives threatening infection in human commonly found in the hospital environment. Our study aimed to isolate bacteriophage against MRSA from the hospital sewage treatment plant and to analyze its efficiency In Vivo in Swiss albino mice model. Sewage sample for the isolation of bacteriophages was collected from SDMH hospital sewage treatment plant in Jaipur. Bacteriophages isolated by the use of enrichment technique and after characterization, isolated phages used to determine phage treatment efficiency in mice. Mice model used to check the safety and suitability of phage application in human need which in turn directly support the use of natural bacteriophage rather than synthetic chemical to kill pathogens. Results show the plaque formation in-vitro and recovery of MRSA infected mice during the experiment. Favorable lytic efficiency determination of MRSA and Salmonella presents a natural way to treat lethal infections caused by Multidrug-resistant bacteria by using their natural host-pathogen relationship.

Keywords: antibiotic resistance, bacteriophages, methicillin resistance Staphylococcus aureus, pathogens, phage therapy, Salmonella typhi

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Authors: Jae Woong Sull

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Lipid levels are related to the risk of cardiovascular diseases. Cholesterol ester transfer protein (CETP) gene is one of the candidate genes of cardiovascular diseases. A total of 2,304 persons were chosen from a Hospital (N=4,294) in South Korea. Female subjects with the CG/GG genotype had a 2.03 -fold (p=0.0001) higher risk of having abnormal HDL cholesterol levels (<40 mg/dL) than subjects with the CC genotype. Male subjects with the CG/GG genotype had a 1.34 -fold (p=0.0019) higher risk than subjects with the CC genotype. When analyzed by body mass index, the association with CETP was much stronger in male subjects with BMI>=25.69 (OR=1.55, 95% CI: 1.15-2.07, P=0.0037) than in male lean subjects. When analyzed by family history of diabetes, the association with CETP was much stronger in male subjects with positive family history of low physical activity (OR=4.82, 95% CI: 1.86-12.5, P=0.0012) than in male subjects with negative family history of diabetes. This study clearly demonstrates that genetic variants in CETP influence HDL cholesterol levels in Korean adults.

Keywords: CETP, diabetes, obesity, polymorphisms

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Authors: Tansa Nisan Gunerhan

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Dementia comes in different forms, including Alzheimer's disease. The most common dementia diagnosis among elderly individuals is Alzheimer's disease. On average, for patients with Alzheimer’s, life expectancy is around 4-8 years after the diagnosis; however, expectancy can go as high as twenty years or more, depending on the shrinkage of the brain. Normally, along with aging, the brain shrinks at some level but doesn’t lose a vast amount of neurons. However, Alzheimer's patients' neurons are destroyed rapidly; hence problems with loss of memory, communication, and other metabolic activities begin. The toxic changes in the brain affect the stability of the neurons. Beta-amyloid and tau are two proteins that are believed to play a role in the development of Alzheimer's disease through their toxic changes. Beta-amyloid is a protein that is produced in the brain and is normally broken down and removed from the body. However, in people with Alzheimer's disease, the production of beta-amyloid increases, and it begins to accumulate in the brain. These plaques are thought to disrupt communication between nerve cells and may contribute to the death of brain cells. Tau is a protein that helps to stabilize microtubules, which are essential for the transportation of nutrients and other substances within brain cells. In people with Alzheimer's disease, tau becomes abnormal and begins to accumulate inside brain cells, forming neurofibrillary tangles. These tangles disrupt the normal functioning of brain cells and may contribute to their death, forming amyloid plaques which are deposits of a protein called amyloid-beta that build up between nerve cells in the brain. The accumulation of amyloid plaques and neurofibrillary tangles in the brain is thought to contribute to the shrinkage of brain tissue. As the brain shrinks, the size of the brain may decrease, leading to a reduction in brain volume. Brain atrophy in Alzheimer's disease is often accompanied by changes in the structure and function of brain cells and the connections between them, leading to a decline in brain function. These toxic changes that accumulate can cause symptoms such as memory loss, difficulty with thinking and problem-solving, and changes in behavior and personality.

Keywords: Alzheimer, amyloid-beta, brain atrophy, neuron, shrinkage

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Authors: Shahid-ul-Islam, Faqeer Mohammad

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The present study was conducted to investigate the effect of annatto, teak and flame of the forest natural dyes on color, fastness, and antimicrobial property of protein based textile substrate. The color strength (K/S) of wool samples at various concentrations of dyes were analysed using a Reflective Spectrophotometer. The antimicrobial activity of natural dyes before and after application on wool was tested against common human pathogens Escherichia coli, Staphylococcus aureus, and Candida albicans, by using micro-broth dilution method, disc diffusion assay and growth curve studies. The structural morphology of natural protein fibre (wool) was investigated by Scanning Electron Microscopy (SEM). Annatto and teak natural dyes proved very effective in inhibiting the microbial growth in solution phase and after application on wool and resulted in a broad beautiful spectrum of colors with exceptional fastness properties. The results encourage the search and exploitation of new plant species as source of dyes to replace toxic synthetic antimicrobial agents currently used in textile industry.

Keywords: annatto, antimicrobial agents, natural dyes, green textiles

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Authors: Mohsina Akhter

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Cancer has become a wide spread disease around the globe and takes many lives every year. Different treatments are being practiced but all have potential side effects with somewhat less specificity towards target sites. Pseudomonas aeruginosa is known to secrete a protein azurin with special anti-cancer function. It has unique cell penetrating peptide comprising of 18 amino acids that have ability to enter cancer cells specifically. Reported function of Azurin is to stabilize p53 inside the tumor cells and induces apoptosis through Bax mediated cytochrome c release from mitochondria. At laboratory scale, we have made recombinant azurin through cloning rpTZ57R/T-azu vector into E.coli strain DH-5α and subcloning rpET28-azu vector into E.coli BL21-CodonPlus (DE3). High expression was ensured with IPTG induction at different concentrations then optimized high expression level at 1mM concentration of IPTG for 5 hours. Purification has been done by using Ni+2 affinity chromatography. We have concluded that azurin can be a remarkable improvement in cancer therapeutics if it produces on a large scale. Azurin does not enter into the normal cells so it will prove a safe and secure treatment for patients and prevent them from hazardous anomalies.

Keywords: azurin, pseudomonas aeruginosa, cancer, therapeutics

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Authors: Tuji Jemal Ahmed, Abdi Mohammed, Geremew Geidare Kailo

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Food security, foodborne illness, and food safety are critical issues that affect the East Harerghe region of Oromia, Ethiopia. Despite the region's potential for agriculture, food insecurity remains a significant problem, with many households experiencing chronic hunger and malnutrition. The region also experiences high rates of foodborne illnesses, including cholera, typhoid, and diarrhea, which are caused by poor hygiene and sanitation practices. Additionally, food safety is a significant challenge, particularly in rural areas, where there is a lack of infrastructure, inadequate food storage facilities, and limited access to information about food safety. There are several factors that contribute to the current situation in the East Harerghe region; firstly, the region is susceptible to natural disasters, for instance, drought, which affects crop yields and livestock production. Secondly, the region also experiences poor infrastructure, which affects the storage and transportation of food, particularly in rural areas. Thirdly, there is a lack of awareness and knowledge on good hygiene and sanitation practices, specifically during food handling, processing, and storage. Fourthly, unitability due to conflict and other forms of land degradation exacerbates food insecurity and malnutrition. Finally, limited access to financial resources and markets commonly affects smallholder farmers by their ability to produce and sell food. To address the current situation in that area, several potential solutions can be implemented; investment in infrastructure is necessary, especially in rural areas, to improve the storage and transportation of food. Education and awareness programs on good hygiene and sanitation practices should target local communities, smallholder farmers, and food vendors. Financial resources and markets should be made more accessible to smallholder farmers, particularly through the provision of credit and improved access to markets. Addressing the underlying causes of conflict and promoting peaceful coexistence can help to reduce displacement and loss of livelihoods. Finally, the enforcement of food safety regulations and the implementation of standards for food processing and storage facilities are necessary to ensure food safety. In conclusion, addressing the challenges of food security, foodborne illness, and food safety in the East Harerghe region requires a coordinated effort from various stakeholders, including the government, non-governmental organizations, and local communities. By implementing the solutions outlined above, the region can improve its food security, prevent foodborne illnesses, and keep food safe for its population. Eventually, building the resilience of communities to shocks such as droughts, floods, and conflict is necessary to ensure long-term food security in the region.

Keywords: foodborne illness, food handling, food safety, food security

Procedia PDF Downloads 77