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Hilsa species are broadly described as largely anadromous fish with a large valuable population size. It is well-known as one of the most critical commercial and occurs in marine, estuarine and riverine environments Hilsa shad, Tenualosa ilisha locally known as Sbour, migrates to the freshwater environment of the Shatt Al-Arab River systems for breeding. It was determined the genetic variation in 70 specimens of T. ilisha from four sites at the Shatt Al-Arab River, the mtDNA cytochrome-b gene was selected as a molecular marker for detecting genetic diversity, origin, and divergence of the population through comparing them with 6 samples from different locations at Indo-Pacific. The AMOVA analysis showed that the variation between groups is 60.97% and 39.02% within population indicating statistically significant P value (P

The lack of cost-effective methods for producing antimicrobial peptides has made it impossible to use their high potential as a new and powerful class of antimicrobial agents. In recent years, extensive research has been conducted to decrease the cost of recombinant proteins production through microorganisms, transgenic animals, and plants. Well-known genetic and physiological characteristics, short-term proliferation, and ease of manipulation make E. coli expression system a valuable host for recombinant proteins production. Expression in periplasmic space is recommended to reduce the inherently destructive behavior of antimicrobial peptides against the expressing microorganism and to decline susceptibility to proteolytic degradation. In this study, a pET-based expression system was used to express buforin I at E. coli periplasmic space, and its antimicrobial, hemolytic, and cell toxicity activities as well as structural stability were evaluated. The hemolysis activity and cytotoxicity of His-tagged buforin I were negligible and its antimicrobial activity did not show a significant difference compared to synthetic buforin I. In addition, in silico investigating of stability of native and His-tagged buforin I showed that RMSF, RMSD and Rg curves had followed a similar trend during 150 ns simulation. Furthermore, evaluating the modelled structures, FTIR and X-ray methods of both peptides indicated an insignificant structural difference. It was concluded that the recombinant buforin I could be a viable alternative to some currently used antibiotics by successfully expressing it in the pET-based expression system.

Antagonistic interactions among different functional guilds of nematodes have been recognized for quite some time, but the underlying explanatory mechanisms are unclear. We investigated responses of tomato (Solanum lycopersicum) to two functional guilds of nematodes - plant parasite (Meloidogyne javanica) and entomopathogens (Heterorhabditis bacteriophora, Steinernema feltiae belowground, and S. carpocapsae) - as well as a leaf mining insect (Tuta absoluta) aboveground. Our results indicate that entomopathogenic nematodes (EPNs): 1) reduced root knot nematode (RKN) infestation belowground, 2) reduced herbivore (T. absoluta) host preference and performance aboveground, and 3) induced overlapping plant defense responses by rapidly activating polyphenol oxidase (PPO) and guaiacol peroxidase (GP) activity in roots, but simultaneously suppressing this activity in aboveground tissues. Concurrently, we investigated potential plant signaling mechanisms underlying these interactions using transcriptome analyses. We found that both entomopathogens and plant parasites triggered immune responses in plant roots with shared gene expression. Secondary metabolite transcripts induced in response to the two nematode functional guilds were generally overlapping and showed an analogous profile of regulation. Likewise, we show that EPNs modulate plant defense against RKN invasion, in part, by suppressing active expression of antioxidant enzymes. Inoculations of roots with EPN triggered an immune response in tomato via up-regulated phenylpropanoid metabolism and synthesis of protease inhibitors (PIs) in plant tissues, which may explain decreased egg laying and developmental performance exhibited by herbivores on EPN-inoculated plants. Furthermore, changes induced in the volatile organic compound (VOC)-related transcriptome indicated that M. javanica and/or S. carpocapsae inoculation of plants triggered both direct and indirect defenses. Our results support the hypothesis that plants 'mistake' subterranean EPNs for parasites, and these otherwise beneficial worms activate a battery of plant defenses associated with systemic acquired resistance (SAR) and/or induced systemic resistance (ISR) with concomitant antagonistic effects on temporally co-occurring subterranean plant pathogenic nematodes and terrestrial herbivores.

Trabajo presentado en el XIII International Scientific Agriculture Symposium AGROSYM 2022, celebrado en Sarajevo (Bosnia y Herzegovina), del 6 al 9 de octubre de 2022, Exopolysaccharides are high molecular weight polymers composed of sugar subunits. Produced exopolysaccharides by lactic acid bacteria (LAB) play a significant role in improvement of organoleptic properties of fermented dairy products such as yogurt. Diversely, the probiotic function of these bacteria and the prebiotic properties of their produced biopolymers promote consumer¿s health. For this purpose, a traditional dairy product known as ¿Kishk¿ was selected. 143 strains of lactic acid bacteria were isolated from Iranian Kishk in Khorasan Province and cultured in formulated MRS mediums with different sugars such as glucose, fructose, sucrose and, lactose (40 g/L) and incubated in anaerobic conditions at 30 and 37°C for 48 hours. The microscopic features of the isolates were assessed and the production of exopolysaccharide in the culture medium was evaluated by disk and ruthenium red methods. The phenol-sulfuric and weight method were used to quantify exopolysaccharide production. Results showed pH of Kishk samples ranged from 3.60 to 4.08 and the average of total mesophilic count and LAB count of samples were 6.50 and 5.89 log CFU/g, respectively. Analysis of data exhibited 79 out of 143 lactic acid bacteria isolates were exopolysaccharide producer and 70% of them were cocci. The average of maximum and minimum production by weight method were 2.61 g/L and 0.08 g/L, respectively. The average of highest and the lowest amount of exopolysaccharide by phenol sulfuric method were measured 1.87 g/L and 0.06 g/L, respectively. This study indicates the potential of exopolysaccharide production by Iranian native species from dairy products.

In this study, the antimicrobial effect of exopolysaccharide (EPS) extracted from Enterococcus strains [E. durans K48 (MT437,248), E. faecium R114 (MT437,249) and E. faecium T52 (MT437,250)] isolated from Kishk was applied against some foodborne pathogenic bacteria using well diffusion and microdilution methods. The antioxidant activity of EPS was also evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging assay and ferric reducing antioxidant power (FRAP) method. The cytotoxicity effect of EPS on human Gingival Fibroblast (HGF) cell line was also assessed. The results obtained by antimicrobial test showed that the most resistant bacteria to the examined EPS was Listeria monocytogenes, and the most susceptible were Staphylococcus aureus and E. faecalis. The results showed that the DPPH inhibitory percentage of EPS (25 mg/mL) from E. durans K48, E. faecium R114, and E. faecium T52 was 53%, 58% and 64%, respectively. EPS from E. faecium T52 displayed the highest reducing power, but statistically, there was no significant difference between the reducing power of EPS T52 and EPS R114 (P ≥ 0.05). The lowest toxicity percentage of EPS k48, EPS T52, and EPS R114 on normal human cell line at a concentration of 0.2 mg/mL was 10%, 15%, and 13%, respectively, which was statistically significant (P < 0.05). The obtained results in the present study indicate that EPS from the examined LAB strains with no in vitro cytotoxicity can be a potential source of natural antioxidant and antibacterial agent to be used in food and pharmaceutical industries., This work was financially supported by a Grant (No. 47333) from Ferdowsi University of Mashhad (Research affairs), Iran.

One of the most promising ways to diagnose cancer especially colorectal cancer (CRC) is to trace its epigenetic events. In this article, a discovery step for detection of methylated DNA markers (MDMs) was performed using SureSelectXT Methyl-Seq in CRC case and control groups in addition to several methylation profiling datasets (GSE48684, GSE53051, GSE77718, GSE101764, and GSE42752). In silico validation of MDMs in colorectal and other cancers was conducted by Lnc2met. MethyLight assay was run on 40 and 47 case and control formalin-fixed paraffin-embedded tissues, respectively and the performance of selected genes were classified by support vector machine (SVM). As a result, 180 regions were identified among all common genes. In addition to SEPT9 and SFRP2, the best three MDM regions were selected from SLC30A10, AKR1B1 and GALNT14. Based on all assays, the best performance was accomplished by SEPT9/AKR1B1 with 98% sensitivity, 99% specificity, 125 positive likelihood ratio, 0.02 negative likelihood ratio and 5074 diagnostic odds ratio. Our results indicate that the AKR1B1/SEPT9 methylation panel detects CRC with a higher performance than SEPT9 methylation, which is a commercial diagnostic test for CRC. However, the creation of a clinically valuable test derived from this study requires performance evaluation in liquid biopsies.

Objective Immunotoxins are one of the most promising ways in therapeutic fields, specially cancer therapy which have a unique toxin-antibody structure, and kill the cancer cell by passing through the cell membrane and entering the target cell. The aim of this study was to engineer and design bovine pancreatic enzyme (RNase A) for to escape from ribonuclease inhibitor (RI) as a toxin segment in designing of immunotoxins used in therapeutic fields. Materials and Methods After bioinformatics investigation using PyMol software, engineering and substitution of target amino acid in wild gene coding RNase A sequence (K91A) were performed. The wild type and recombinant proteins in E.coli (BL21(D3) strain) were expressed and refolded, and then approved using SDS-PAGE. The purification of produced proteins was conducted using an immobilized-metal affinity chromatography for His-tagged proteins. Results The results showed that the protein concentrations were 3 and 2.2 g/L for wild RNase A and recombinant enzyme. The hydrolysis activity measurement of the produced enzymes, in presence of different concentrations of RI, indicated that recombinant variant had lower sensitivity and higher catalytic activity in comparison with the wild enzyme. The results obtained from enzyme activity of recombinant enzyme compared to the wild enzyme, in presence or absence of RI, demonstrated that the interaction effect between RNase A and RI can be disrupted by manipulation of amino acids involved in this interaction. Conclusion In respect to the current results, it was concluded that RNase A enzyme can be used as a promising drug in engineering and production of immunotoxins.

Sheep satellite cells more than satellite cells of the rat and mouse are similar to human satellite cells. These cells are widely used in the modeling and treatment of diseases like heart insufficiency, neurological diseases, muscular dystrophy, cerebral cell transplantation for the treatment of migraines, screening, and the production of new drugs. This study was aimed to isolate and culture primary satellite cells (PSCs) obtained from sheep fetus, and perform clonal expansion of transfected PSCs. Skeletal muscle tissues of hind limbs were collected from sheep fetuses obtained from a local abattoir. After enzymatic digestion, flasks were replaced after 3 hours to isolate non-myogenic cells, such as fibroblasts. After six days, the cells were differentiated to myoblasts. Using a differentiation medium containing the horse serum, myotube cells were observed in the flask, indicating that the cultured cells were satellite cells. The mRNA expression of the PAX7 gene was used to confirm the presence of satellite cells. In addition, the results showed that satellite cells grow in a culture medium containing 5% FBS without differentiation, while 10% FBS initiates their differentiation.

One of the most effective methods for increasing the antimicrobial activity of a substance is to combine it with one or more other antimicrobial agents. The aim of the present study was to evaluate the antimicrobial effect of buforin I and nisin alone and investigate the synergistic action of these compounds against the most important food spoilage microorganisms in clouding B. subtilis, S. epidermidis, L. innocua, E. coli, S. Enteritidis, A. oryzae, R. glutinis and G. candidum. The results of MIC and MBC/MFC examinations showed that buforin I had higher antimicrobial activity than nisin on all the microbial strains used in this study (p≤0.5). E.coli was the most resistant to both antimicrobial agents, while Listeria innocua and Staphylococcus epidermidis were the most sensitive to nisin and buforin I, respectively. The results of synergistic interaction between buforin I and nisin indicated that the combination of buforin I and nisin on B. subtilis, S. epidermidis and A. oryzae showed synergistic effect, while it had no effect on S. Enteritidis and Geotrichum candidum. The combination of buforin I and nisin showed partial synergistic effect on Listeria innocua, Escherichia coli, Rhodotorula glutinis. Assessment of viability of the microorganisms under the antimicrobial agents alone and in combination with each other at MICs and FICs indicated that use of these antimicrobial agents in combination enhances antimicrobial activity at lower concentrations of both agents. The present study investigated the antimicrobial properties of buforin I against food spoilage microorganisms for the first time and suggests that its use alone or in combination with nisin may provide a clear horizon for the application of antimicrobial peptides as natural preservatives. Thus, the combination of antimicrobial peptides and traditional antimicrobial food preservative could be a promising option for the prevention of contamination, spoilage, and infestation of food and beverage products.

To investigate the effects of dietary supplementation with industrial vinegar (IV) and waste date vinegar (WDV) on growth performance, intestinal morphology, and immune response of broilers, five hundred Ross 308 chickens were randomly divided into 50 cages in a 42-day breeding period. The experiment consisted of ten treatments with five replications in each treatment including; control 1 (standard feed without WDV or IV), control 2 (control 1 + 2% water into the feed), 1, 2, and 3% of WDV and 2% industrial vinegar (IV) into the feed and 0.5, 1, and 1.5% of WDV and 1% industrial vinegar (IV) into the water. One chicken from each replicate was slaughtered on days 24 and 42 to investigate gastrointestinal tissue growth as well as intestinal morphology. The results showed that growth performance was not affected by treatments in any period. The height and width of the villus in the treatments containing 1% of WDV into the water and 2% of WDV into the feed increased with time compared to the control treatments. On day 42 of the experiment, the crypt depth was also higher in the treatment containing 1% of WDV into the feed compared to the other treatments. The ileal coliforms were also affected by WDV addition into the feed at 42 days of age compared to other groups (P < 0.05). The highest amount of IgM and IgG were found to levels of 2% and 1% of WDV into the diet, respectively (P < 0.05). However, SRBC, ND titter was not affected by treatments. Also, no difference was found between industrial vinegar in water or feed in most of the studied parameters. The results of this study showed that supplementation of the diet with WDV had a positive effect on intestinal morphology and immune system of broilers compared to industrial vinegar.

Objective Salmonella is a Gram-negative, anaerobic, flagellated bacterium of Salmonella type in Enterobacteriaceae family, which is known as a zoonosis infectious agent. In recent years, different types of antibiotics have been used to overcome the Salmonella infection; however, the increasing number of antibiotic-resistant bacteria, the provision of alternative methods seems be necessary. Specific immunoglobulin Y (IgY) originated from egg yolk causes passive immunity in the fetus, and extracting these antibodies from egg yolk is cheaper and more feasible than other antibodies. Numerous studies have shown that the egg yolk IgY has immunogenic function against a wide range of bacterial and viral infections in mammals. The purpose of this study was to produce specific immunoglobulin Y against Salmonella typhimurium and Salmonella enteritidis bacteria in laying hens. Materials and Methods In the current study, six pullets were inoculated and boosted with one milliliter of killed Salmonella typhimurium and Salmonella enteritidis. Injections were done fortnightly until one week before egg production. After breast muscle injection of chickens, the eggs were collected, and the IgY was extracted from yolk by the Polson method with polyethylene glycol 6000. The produced IgY was approved by SDS-PAGE method. The specific indirect ELISA done by specificity test for purified IgY also approved the specific IgY against both Salmonella typhimurium and Salmonella enteritidis. Results The results of SDS - PAGE demonstrated the presence of two protein bands of 27 and 67 kDa representing light and heavy chains of IgY, respectively. The specific indirect ELISA method proved that the specific IgY against Salmonella typhimurium and Salmonella enteritidis was produced in the egg yolk. Conclusions It might be possible to administer the egg yolk containing specific IgY against both Salmonella typhimurium and Salmonella enteritidis, to induce passive immunity for preventing some pathogenic diseases in calves such as diarrhea.

Background: Due to the heating of amino acids with edible oils to high temperatures, different amounts of acrylamide are produced. Objective: The purpose of this study was to compare the level of acrylamide in the tahdig of bread and tahdig of potato prepared with the common liquid and solid edible oils, including sunflower, corn, canola, frying oil and solid oils. Methods: The tahdig of bread and potato was prepared under the same temperature and time with different oils. Acrylamide isolation was performed on a solid-phase extraction (SPE) cartridge and acrylamide was determined using High-performance liquid chromatography (HPLC). Results: The highest amount of acrylamide was obtained with sunflower oil in the tahdig of potato (194.091 mg/Kg) and the lowest amount of acrylamide was obtained with solid oil in the tahdig of bread (48.54 mg/Kg). For all the oils, the acrylamide content of the tahdig of potato was higher than bread. Conclusion: This study clearly demonstrated the involvement of the kind of oils in the formation of acrylamide in the tahdig of bread.

The novel coronavirus (SARS-CoV-2) has without a doubt escalated to become a global crisis. Taking into consideration our limited knowledge regarding the virus, all the efforts to provide better understanding or explore the solutions are highly welcomed. In this article, 88 conventional drugs, 16 vitamins, and 63 natural (plant) compounds were chosen to perform a binding simulation with the reported COVID-19 main protease (Mpro) in search for probable inhibitors. Based on docking results, various vitamins (B9, A, K, and E vitamins) exhibited a significantly strong interaction with the studied receptor which might refer to the importance of these supplements in daily diets. Additionally, the strong ligand-protein interactions of some conventional drugs such as Pleconaril, Adefovir dipivoxil, and Stavudine in addition to plant-based compounds such as Curcumin (Curcuma longa), Anolignan A (Anogeissus acuminata), and Phyllamyricin B (Phyllanthus myrtifolius) render these compounds promising and recommended for further studies.

Recently, ‘solid tumor biopsies’ have been challenged by the emergence of ‘liquid biopsies’, which are aimed at the isolation and detection of circulating cell-free tumor DNA (ctDNA) in body fluids. Here, we developed and optimized a method for selective capture of ctDNA on magnetic beads (SCC-MAG) for mutation detection in plasma of patients with colorectal cancer (CRC). Blood and tissue samples from 28 CRC patients were included for the detection of KRAS mutations. For the tissue samples, mutation analysis was conducted by high resolution melting (HRM) analysis and sequencing. For the SCC-MAG method, ctDNA was isolated from 200 µl plasma from patients with a mutant KRAS gene. For comparison, ctDNA extraction was carried out using a silica membrane-based method, after which mutations were detected using Intplex allele-specific PCR. The mean ctDNA integrity index in plasma samples of cancer patients was 1.03, comparable with that of silica membrane-derived ctDNA (1.011). Notably, the limit of detection for the SCC-MAG approach was lower than that of the silica membrane method and measured 2.25 pg/ml ctDNA in plasma. Our analyses showed that while the silica membrane-based approach was capable of collecting ctDNA from two out of six CRC patient samples (average Cq 34.23), the SCC-MAG captured ctDNA from all samples with an average Cq of 29.76. We present a robust, reproducible, and highly sensitive method for the analysis of mutation statuses in liquid biopsies. The SCC-MAG method can readily be applied to any nucleic acid target for diagnostic purposes upon careful design of the specific capture probes, and can be multiplexed by several probes to identify multiple targets.

This study was performed to investigate the effects of dietary threonine (Thr). level on performance, metabolizable energy, intestinal morphology, and immune system in coccidian–infected broiler chickens. The diets contained: 88%, 100% (Non challenged (NC) and challenged control (PC)), 112%, 124%, and 136% of Thr requirement according to Cobb 500 recommendation and fed during grower (pre challenged) and finisher (post challenged) periods. On d 23 (end of grower period), each bird received 0.5 mL of distilled water or received around 24000 sporulated oocysts. On d 23 and 31, one bird per replicate was slaughtered to measure the performance criteria. Mean dietary apparent metabolizable energy corrected for nitrogen (AMEn) and digestible energy were greater in NC birds than the challenged birds fed on 88% or 100% Thr diets. Feed intake and blood parameters were not significantly influenced by increasing levels of Thr in the diet. Compared with unchallenged birds with coccidia (NC), the growth performance, morphological parameters (not crypt depth), and immune responses decreased (P < 0.05) in the birds (PC) that were challenged with coccidia, and oocyte numbers were enhanced. Growth performance, jejunal morphology, and immune responses improved and oocyte count decreased in coccidian- challenged birds fed on the diets with greater levels of Thr (P < 0.05). Feeding the challenged birds with the diet containing greater levels of Thr improved (P < 0.05) their growth performance, morphology, and immune responses and decreased oocyte number. The birds fed on the diet with 124% Thr demonstrated a similar response as the NC birds. Increased diet Thr level linearly increased average daily gain and decreased feed conversion ratio in the grower and the whole experimentation periods. The AMEn and digestibility of crude protein were enhanced linearly Thr level increased in coccidian-challenged birds. It is concluded that diets containing 124% of Thr recommendation led to the greatest efficacy on the intestinal immuneresponse and normal growth maintenance of the birds contaminated with coccidia.

Lactoferrin is one of the most important components in milk. Lactoferrin is an 80 kDa iron-binding glycoprotein that involves a diverse range of biological activities. However, the antimicrobial properties are its most widely studied function. The aim of this study was to assay the antimicrobial and antibiofilm activities of bovine lactoferrin against five of the most important foodborne pathogenic bacterial strains including Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Listeria monocytogenes and Listeria innocua, in planktonic and biofilm forms. Bovine lactoferrin was purchased, MIC and MBC were determined for the mentioned bacterial strains and the effect of lactoferrin concentrations on biofilm formation was evaluated using microtiter plate method. Based on the results, lactoferrin had a greater effect on gram-positive bacteria, L. monocytogenes and L. innocua were the most sensitive strains. MIC for L. monocytogenes, L. innocua, S. aureus and P. aeruginosa was 6, 6, 8 and 14 mg/mL, respectively, Nevertheless, MIC for gram negative E. coli was not observed, even at 16 mg/mL. The result demonstrated that MBC for L. monocytogenes, L. innocua and S. aureus were 10, 12 and 16 mg/mL, respectively. However, MBC for P. aeruginosa and E. coli were not observed for the lactoferrin concentrations evaluated. The highest biofilm formation inhibitory effects of bovine lactoferrin were against L. monocytogenes and L. innocua. To our knowledge, this is the first report on the antibiofilm activity of bovine lactoferrin against foodborne pathogenic bacteria. Our findings suggest lactoferrin to possibly use as an anti-biofilm agent in both food and pharmaceutical industries.

Sheep has been an essential source of food to the inhabitants of the Iranian plateau for centuries. Furthermore, this geographic area is considered the original place of sheep domestication. Phylogenetic studies are highly important in understanding the evolutionary relationships among species. This understanding assists in decision making and planning for genetic resources conservation programs. Analyzing sequences of mitochondrial genome regions provides more reliable evidence regarding the genetic diversity and evolutionary origin of the species, due to the high rate of mitochondrial genome evolution compared to the nucleus. The aim of this study was to use the sequence of mitochondrial control region to investigate the phylogenetic relationship and genetic distances between some domestic sheep breeds of Iran and other Middle Eastern countries. For this purpose, blood samples were collected from Zel, Dalagh, and Mehrabani sheep breeds. After DNA extraction, polymerase chain reaction (PCR) was carried out using specific primers for the targeted mitochondrial genome. Then, PCR products were purified and a standard sequencing was performed. Sequences obtained from this study were compared with other (NCBI) registered sequences of Middle Eastern breeds. Analysis of phylogenetic tree of the main haplotypes of sheep revealed that Zel breed was grouped in the haplotype B with other thin-tailed breeds, such as Karayaka and Sakiz. On the other hand, Mehrabani sheep breed was placed in the haplotype A along with other Middle Eastern fat-tailed breeds such as Naeimi and Saidi. This study provided additional proof for the use of control region sequence as a precise method of genetic distance estimation among sheep breeds.

KRAS mutation is responsible for 40–50% of colorectal cancers (CRCs). RNA-seq data and bioinformatics methods were used to analyze the transcriptional profiles of KRAS mutant (mtKRAS) in comparison with the wild-type (wtKRAS) cell lines, followed by in-silico and quantitative real-time PCR (qPCR) validations. Gene set enrichment analysis showed overrepresentation of KRAS signaling as an oncogenic signature in mtKRAS. Gene ontology and pathway analyses on 600 differentially-expressed genes (DEGs) indicated their major involvement in the cancer-associated signal transduction pathways. Significant hub genes were identified through analyzing PPI network, with the highest node degree for PTPRC. The evaluation of the interaction between co-expressed DEGs and lncRNAs revealed 12 differentially-expressed lncRNAs which potentially regulate the genes majorly enriched in Rap1 and RAS signaling pathways. The results of the qPCR showed the overexpression of PPARG and PTGS2, and downregulation of PTPRC in mtKRAS cells compared to the wtKRAS one, which confirming the outputs of RNA-seq analysis. Further, significant upregualtion of miR-23b was observed in wtKRAS cells. The comparison between the expression level of hub genes and TFs with expression data of CRC tissue samples deposited in TCGA databank confirmed them as distinct biomarkers for the discrimination of normal and tumor patient samples. Survival analysis revealed the significant prognostic value for some of the hub genes, TFs, and lncRNAs. The results of the present study can extend the vision on the molecular mechanisms involved in KRAS-driven CRC pathogenesis.

Early, specific, and sensitive detection methods of COVID-19 are essential for force stopping its worldwide infection. Although CT images of the lung and/or viral RNA extraction followed by real-time reverse-transcriptase-polymerase chain reaction (rRT-PCR) are widely used; they have some limitations. Here, we developed a highly sensitive magnetic bead-based viral RNA extraction assay followed by rRT-PCR.Case group included oropharyngeal/nasopharyngeal and blood samples from 30 patients diagnosed positive by PCR test for COVID-19 and control group included 30 same samples from COVID-19 negative PCR test individuals. RNA was extracted, using viral RNA extraction kit as well as using our hand-made capture bead-based technique. A one-step cDNA synthesis and Real Time PCR was conducted. A two-step comparison of the different viral RNA extraction methods for oropharyngeal/nasopharyngeal and blood samples was performed. Student t-test was applied with aIn the case group, all 30 mucosal samples extracted either with viral RNA extraction kit or with beads-based assay were COVID-19 positive although in the latter category, Cqs were much lower. Although 43% of plasma samples extracted by bead-based method were found to be positive but no plasma samples extracted with column-based kit were detected positive by Real Time PCR.Bead-based RNA extraction method can reduce RNA loss by its single-tube performance and enhance the test sensitivity. It is also more sensitive to lower viral loads as shown in the detection of blood samples and the lower Cqs of mucosal samples.

Clostridium perfringens-induced necrotic enteritis (NE) is an economically important disease of broiler chickens. The present study evaluated the effect of C. perfringens on the intestinal histomorphometry, enteric microbial colonization, and host immune responses using 3 experimental NE reproduction methods. The experimental groups consisted of 1) unchallenged Control diet (corn-soybean meal), 2) Control diet + Eimera inoculation at d 11 followed by C. perfringens challenge at d 15 (ECp), 3) Wheat-based diet + C. perfringens challenge (WCp), and 4) Wheat-based diet + Eimeria inoculation followed by C. perfringens challenge (WECp). The results showed that chickens receiving ECp and WECp had reduced (P < 0.05) bird performance coupled with enteric gross lesions and epithelial damage at d 17 and 24 of age compared to unchallenged control birds. These ECp and WECp administered birds also had increased (P < 0.05) ileal colonization by clostridia and E. coli at d 17 and 24, while the resident Lactobacillus counts were reduced (P < 0.05) at d 24 of age. Furthermore, at d 24, jejunal transcription of IL-6, IL-10, annexin-A1 and IL-2 genes was upregulated (P < 0.05) in the ECp group, whereas the transcription of TNF receptor associated factor (TRAF)-3 gene was increased (P < 0.05) in WECp treated birds when compared to unchallenged control group. Additionally, stimulation of chicken splenocytes and cecal tonsilocytes with virulent C. perfringens bacilli or their secretory proteins resulted in a higher (P < 0.05) frequency of T cells and their upregulation of MHC-II molecule, as determined by flow cytometry. These findings suggest that C. perfringens, while inducing epithelial damage and changes in microbiota, can also trigger host immune responses. Furthermore, NE reproduction methods using coccidia with or without the wheat-based dietary predisposition seem to facilitate an optimal NE reproduction in broiler chickens and thus, may provide better avenues for future C. perfringens research.

This study aimed to monitor the effect of including rumen-protected L-carnitine (Carneon 20 Rumin-Pro, Kaesler Nutrition GmbH, Cuxhaven, Germany) in the transition diet on the productive and metabolic responses of multiparous high-producing Holstein dairy cows. Thirty-two multiparous cows were allocated in a completely randomized design to receive the same diet plus 60 g fat prill containing 85% palmitic acid (control, n = 16) or 100 g rumen-protected L-carnitine (RLC, n = 16); at 28 days before expected calving until 28 days in milk (DIM). Fat prill was included in the control diet to balance the palmitic acid content of both experimental diets. Milk production over the 28 DIM for the control and RLC groups was 46.5 and 47.7 kg, respectively. Milk fat content tended to increase upon rumen-protected L-carnitine inclusion (p = 0.1). Cows fed rumen-protected L-carnitine had higher fat- and energy-corrected milk compared with the control group. Pre- and post-partum administration of L-carnitine decreased both high- and low-density lipoprotein concentrations in peripheral blood of post-partum cows. The results of this study indicated that the concentration of triglycerides and beta-hydroxybutyrate was not significantly different between the groups, whereas the blood non-esterified fatty acid concentration was markedly decreased in cows supplemented with L-carnitine. Animals in the RLC group had a significant (p < 0.05) lower blood haptoglobin concentration at 7 and 14 DIM than the control. Animals in the RLC group had a lower concentration of blood enzymes than those of the control group. The mRNA abundance of Toll-like receptors 4, cluster of differentiation 14, and myeloid differential protein 2 did not significantly change upon the supplementation of L-carnitine in the transition diet. In summary, the dietary inclusion of RLC improved dairy cow's performance during the early lactation period. Greater production, at least in part, is driven by improved energy utilization efficiency and enhanced metabolic status in animals during the periparturient period.

DNA methylation is a fundamental epigenetic process and have a critical role in many biological processes. The study of DNA methylation at a large scale of genomic levels is widely conducted by several techniques that are next-generation sequencing (NGS)-based methods. Methylome data revealed by DNA methylation next-generation sequencing (mNGS), should be always verified by another technique which they usually have a high cost. In this study, we offered a low-cost approach to corroborate the mNGS data. In this regard, mNGS was performed on 6 colorectal cancer (case group) and 6 healthy individual colon tissue (control group) samples. An R-script detected differentially methylated regions (DMRs), was further validated by high resolution melting (MS-HRM) analysis. After analyzing the data, the algorithm found 194 DMRs. Two locations with the highest level of methylation difference were verified by MS-HRM, which their results were in accordance with the mNGS. Therefore, in the present study, we suggested MS-HRM as a simple, accurate and low-cost method, useful for confirming methylation sequencing results.

Computational prediction of signal peptides is one of the most important steps in genetic engineering experiments. The periplasmic expression cause the reducing in the inherent destructive behavior of Bofurin I against its host and also reducing its susceptibility to proteolytic degradation. In order to predict the best signal peptides for expression of Buforin I in E. coli, 103 signal sequences were retired from signal peptide databases. Since the purpose of this study was to introduce the optimal signal peptides for periplasmic expression, first, sub-cellular localization site of signal peptides was analyzed. Then, n, h, and c regions of signal peptide, signal peptide probability and physico-chemical features were investigated. Base on the results, MalE, hofQ, papK, ugpB, zraP, and sfmC were introduced as the best signal peptides. For increasing the half-life of mRNA and the increasing the stability of the mRNA against exonuclease activity, secondary structures of mRNA including Shine-Dalgarno, untranslated region of ompA, start codon, signal peptide and sequences of Buforin I were analyzed. Based on the total free energy pilot evaluated and mRNA conformations, papK seemed more appropriate than the rest of the signal peptides. The obtained result of this study can be used for design the periplasmic expression constructs.

Background:Thanatin is the smallest member of Beta-hairpin class of cationic peptide derived from insects with vast activities against various pathogens.Objective:n this study, the antimicrobial activity of this peptide against some species of human bacterial pathogens as well as its toxicity on NIH cells were evaluated.Method:Thanatin DNA sequence was cloned into pcDNA3.1+ vector and transformed into a DH5α bacterial strain. Then the recombinant plasmids were transfected into HEK-293 cells by calcium phosphate co-precipitation. After applying antibiotic treatment, the supernatant medium containing thanatin was collected. The peptide quantity was estimated by SDS-PAGE and GelQuant software. The antimicrobial activity of this peptide was performed with Minimum Inhibitory Concentration (MIC) method. In addition, its toxicity on NIH cells were evaluated by MTT assay.Results:The peptide quantity was estimated approximately 164.21 µmolL-1. The antibacterial activity of thanatin was estimated between 0.99 and 31.58 µmolL-1 using MIC method. The result of cytotoxicity test on NIH cell line showed that the peptide toxicity up to the concentration of 394.10 µmolL-1 and for 48 hours, was not statistically significant from negative control cells (P>0.05). The antimicrobial assay demonstrated that thanatin had an antibacterial effect on some tested microorganisms. The results obtained in this study also showed that thanatin had no toxicity on mammalian cell lines including HEK293 and NIH.Conclusion:Antimicrobial peptides such as thanatin are considered to be appropriate alternatives to conventional antibiotics in treating various human pathological diseases bacteria.

Gamma amino butyric acid (GABA) is used as drugs, food ingredients, and dietary supplements. l-glutamate is converted to GABA by the decarboxylation reaction, which is catalyzed by the glutamate decarboxylase (GAD). Escherichia coli is widely being used to express proteins. However, without appropriate signal peptide, it cannot be applied for secretory proteins. Selecting a suitable signal peptide (SP) is a critical step in the secretory production of different proteins. In silico identification of suitable SP is a reliable and cost-effective alternative to experimental approaches. In previous studies, the localization of proteins was not considered and the SPs of periplasmic, membranes and extracellular were compared. Therefore, this study aimed to predict the best SP for the expression of recombinant GAD in the outer membrane of E. coli only. Also, we compared twelve servers to evaluate protein localization, solubility, and secretory pathway. In the present study, 127 SPs were taken from the Signal Peptide database. The localization site, physico-chemical properties, location of cleavage sites, regions and D-score of them were determined by ProtComp, ProtParam, and SignalP 3.0 and 4.1 servers, respectively. To rank SPs based on the secretion properties, PRED-TAT and SignalP 5.0 webservers were used. Based on the results, the localization site of 13 SPs was in the outer membrane of E. coli. Among them, the most suitable candidates seemed to be torT with a reasonably high D-score, aliphatic index, and GRAVY, followed by ccmH and then pspE. TorT accelerates GAD scale-up production and might be useful in future experimental research.

In the present study, a novel chimeric peptide was derived from camel lactoferrin designed with a considerable anti-HCV activity and its neutralization mechanism was predicted by molecular modelling tools. A novel anti-HCV peptide derived from camel lactoferrin (cLF36) was designed and expressed it recombinantly in HEK-293-T cells. Anti-viral activity of this peptide was evaluated against hepatitis C virus by Real-time PCR assay in vitro. Finally, to have a better insight into the mode of action of peptide on HCV entry inhibition, we examined the interaction of cLF36 with envelope glycoprotein E2 by molecular dynamic simulation. This chimeric peptide had significant inhibitory effects on both HCV entry (44 µg/mL) and viral replication (88 µg/mL) under in vitro (p > 0.01). Moreover, cLF36 peptide was not toxic to HEK cells as a normal cell at twofold of its anti-viral concentrations for HCV entry and even at concentrations as high as 250 µg/mL exhibited minimal hemolysis (2.5%) against human RBCs (red blood cells). The results of in silico analysis showed that cLF36 interacted with β-sandwich and front layer of E2 protein as two potential CD81 binding sites. We generated and characterized a new camel lactoferrin derived HCV inhibitors. This peptide blocked HCV entry and also intracellular HCV replication in cell culture experiment.

Bovine leukemia virus (BLV) is the etiologic agent of enzootic bovine leucosis (EBL) for the bovine host. In this study to examine gene expression changes in the manifestation of the EBL malignancy, four pooled RNA samples (three RNAs in each sample) were applied for transcriptome sequencing using RNA-seq technique. Differential expression analysis was done to compare the infected bovine group with the healthy bovine group using DESeq2 package in R software. Furthermore, functional gene ontology (GO) term and KEGG pathway enrichment analysis were stablished using the DAVID online database to identify involved GO terms and pathways in the host response to BLV infection. Our results suggested that 371 up- and 72 downregulated genes were involved in EBL with statistically significant threshold log2foldchange (LFC) = 1 and false discovery rate (FDR)

Growth hormone has mitotic and anti-apoptotic effects which may increase proliferation and transformation of cells when it is expressed aberrantly. This study investigated the effects of resistance training and growth hormone injection on circulating IGF-1, IGFBP-3 levels and IGF-1/IGFBP-3 ratio in male Wistar rats. Thirty-two male Wistar rats were randomly assigned to a control group (C, n = 8), a resistance training group (RT, n = 8), a growth hormone injection group (GI, n = 8) and a resistance training + growth hormone injection group (RG, n = 8). The resistance training protocol comprised of climbing a ladder (5 days/week, 3 sets/5 reps) while carrying a weight suspended from the tail. The growth hormone (2 mg/kg/day, 5 days/week) was injected before an exercise session. Serum IGF-1, IGFBP-3 levels, and IGF-1/IGFBP-3 ratio were measured after 8 weeks. One-way ANOVA analysis was used for comparison of serum IGF-1 and IGFBP-3 levels between groups. Serum IGF-1 levels and IGF-1/IGFBP-3 ratio signifi cantly decreased, but serum IGFBP-3 levels showed no significant change in the RT group compared to the C group. Also, both serum IGF-1 and IGFBP-3 levels and IGF-1/IGFBP-3 ratio in GI and RG groups significantly increased compared to the other groups. In conclusion, resistance training decreases serum IGF-1 levels and/or IGF-1/IGFBP-3 ratio in normal condition. On the other hand, the growth hormone injection with and without the resistance training increases serum IGF-1 levels and IGF-1/IGFBP-3 ratio which could be noted as a condition with a higher risk of neoplasm.

1. The objective of the study was to investigate the influence of maternal and parent of origin effects (POE) on genetic variation of Iranian native fowl on economic traits. 2. Studied traits were body weights at birth (BW0), at eight (BW8) and 12 weeks of age (BW12), age (ASM) and weight at sexual maturity (WSM), egg number (EN) and average egg weight (AEW). 3. Several models, including additive, maternal additive genetics, permanent environmental effects and POE were compared using Wombat software. Bayesian Information Criterion (BIC) was used to identify the best model for each trait. The chance of reranking of birds between models was investigated using Spearman correlation and Wilcoxon rank test. 4. Based on the best model, direct heritability estimates for BW0, BW8, BW12, ASM, WSM, EN and AEW traits were 0.05, 0.21, 0.23, 0.30, 0.39, 0.22 and 0.38, respectively. Proportion of variance due to paternal POE for BW8 was 4% and proportion of variance due to maternal POE for BW12 was 5%. 5. Estimated maternal heritability for BW0 was 0.30 and for BW8 and BW12 were 0.00 and 0.01, respectively, which shows that maternal heritability was reduced by age. 6. Based on the results, considering POE for BW8 and BW12 and maternal genetic effects for BW0 improved the accuracy of estimations and avoid reranking of birds for these traits.

The production of polysaccharides derived from lactic acid bacteria (LAB) can be a valuable alternative to current polysaccharides. In this study, eight samples of Kishk (traditional dairy product) were collected in sterile conditions and directly cultured on MRS agar medium. Following purification and examination of microscopic, macroscopic characteristics and doing biochemical tests, 143 isolates were selected, and the production of exopolysaccharide (EPS) was investigated by the ruthenium red and disc methods. The EPS production of 79 isolates was confirmed. Total carbohydrate of EPS was determined by the phenol sulfuric acid method. Finally, Enterococcus durans K48, Enterococcus faecium R114, and Enterococcus faecium T52 strains were selected as the best EPS producers. The optimization of EPS production was then performed using Central Composite Design (CCD) and Response Surface Methodology (RSM) approaches. Optimization plots showed the highest EPS yield for E. durans K48, E. faecium R114, and E. faecium T52 occurred at 38.4, 37.4 and 36.7 °C and pH of 5.9, 5.6 and 5.8, respectively, cultured in a Sucrose-MRS medium. Under optimal conditions, the maximum predicted and actual production of EPSs for the examined isolates were 3.18, 3.21, 2.99 and 3.02, 3.15, 3.15, g L, respectively. As determined by Gel Permeation Chromatography, the EPSs molecular weights were in the range of 2.93 × 10 to 3.52 × 10 Da. Analysis of the component monosaccharides by HPLC showed that all three tested EPSs were heteropolysaccharides. After appropriate evaluation, the EPSs produced by native species of LAB isolated from Iranian Kishk could be of interest for industrial applications or as functional food ingredients., This work was financially supported by a grant (No. 3/47333) from the Ferdowsi University of Mashhad (Research Affairs), Iran.

We investigated responses of tomato to two functional guilds of nematodes - plant parasite (Meloidogyne javanica) and entomopathogens (Heterorhabditis bacteriophora, Steinernema feltiae belowground, and S. carpocapsae) - as well as a leaf mining insect (Tuta absoluta) aboveground. Our results indicate that entomopathogenic nematodes (EPNs): 1) induced plant defense responses, 2) reduced root knot nematode (RKN) infestation belowground and 3) reduced herbivore (T. absoluta) host preference and performance aboveground. Concurrently, we investigated the plant signaling mechanisms underlying these interactions using biochemical and transcriptome analyses. We found that both entomopathogen and parasite triggered immune responses in plant roots with shared gene expression. Tomato plants responded similarly to presence of RKN or EPN in the root zone, by rapidly activating polyphenol oxidase (PPO) and guaiacol peroxidase (GP) activity in roots, but simultaneously suppressed this activity in aboveground tissues. We quantified changes in gene expression in tomato that may play essential roles in defense response to RKN, which were also coincidentally triggered by EPN. Overall, EPN inoculation directly mediated enhanced plant defense and reduced subsequent RKN infection. Likewise, we show that EPNs modulate plant defense against RKN invasion. Inoculation of tomato roots with EPNs belowground reduced both host preference and performance of the aboveground herbivore, T. absoluta. Inoculations of roots with EPN also triggered an immune response in tomato which could explain an observed decrease in egg laying and developmental performance exhibited by herbivores on EPN-inoculated plants. Our results support the hypothesis that subterranean EPNs activate a battery of plant defenses associated with systemic acquired resistance (SAR) and/or induced systemic resistance (ISR) with antagonistic effects on temporally co-occurring subterranean plant pathogenic nematodes a

Cationic antimicrobial peptides are being developed as a promising class of antimicrobial sub-stances. The introduction of a new antibiotic component requires a comprehensive study of its properties so that it can be relied upon to continue laboratory procedures and clinical trials on laboratory animals or human volunteers. Antimicrobial activity of buforin I was evaluated against 15 of the most important pathogenic bacterial and fungal strains. This was followed by assessing anti-biofilm activity, time-dependent inhibitory, thermal stability, plas-ma stability, hemolysis, and cytotoxic activities. The range of obtained MICs was between 4 and 16 μg/mL. The most resistant and most sensitive microbial strains were S. salivarius and C. perfringens, respectively. Buforin I not only inhibited biofilm formation, but also showed a high biofilm radiation activity. Buforin I was stable in human plasma and also at different temperatures including 40, 60, and 80 °C. Although no significant anti-cancer properties were observed for buforin I, the lack of cytotoxicity as well as the lack of hemolytic activity confirm its safety. The high therapeutic index indicated that buforin I has a considerable pharmaceutical potential and can be a reasonable candidate to replace antibiotics or administered in combination with antibiotics to increase the effectiveness as well as reduce the dose of antibiotics.

There is a continuing need to prevent the increasing use of common antibiotic and find the replacement to combat the drug/antibiotic resistant bacteria such as antimicrobial peptides (AMPs) such as thanatin peptide. In this study, recombinant thanatin peptide was expressed in the HEK293 cell line. Then the antimicrobial properties of this peptide on some poultry and farm animal’s pathogen strains were assessed. The thermal-stability of thanatin was predicted in various temperatures through in silico analysis. Afterwards, according to Minimum Inhibitory Concentration (MIC) results, Escherichia coli and Pseudomonas aeruginosa were chosen to test the hypothesis of LptA/LptD–thanatin interaction, computationally. Relative amino acid sequences and crystallography structures were retrieved and missed tertiary structures were predicted. The interaction of thanatin with LptA and LptD of Escherichia coli and Pseudomonas aeruginosa were analyzed subsequently. The antibacterial activity of thanatin peptide was evaluated between 6.25 and 100 μg/mL using minimum inhibitory concentration. Also, the amounts of minimum bactericidal concentrations (MBC) were between 12.5 and 200 μg/mL. The bioinformatics analysis followed by the in vitro assessment, demonstrated that thanatin would be thermally stable in the body temperature of poultry and farm animals. Thanatin could penetrate to the outer membrane domain of LptD in Escherichia coli and it could block the transition path of this protein while the entrance of LptD in Pseudomonas aeruginosa was blocked for thanatin by extra residues in comparison with Escherichia coli LptD. In addition, the quality of interaction, with regard to the number and distance of interactions which leads to higher binding energy for thanatin and LptD of Escherichia coli was much better than Pseudomonas aeruginosa. But the site and quality of interaction for thanatin and LptA was almost the same for Escherichia coli and Pseudomonas aeruginosa. Accordingly, thanatin can prevent the assembly of LptA periplasmic bridge in both pathogens. The antibacterial and thermal stability of the thanatin peptide suggested that thanatin peptide might serve as a natural alternative instead of common antibiotics in the veterinary medicine. The outcome of this in silico study supports the MIC results. Therefore, a probable reason for different level of activity of thanatin against Escherichia coli and Pseudomonas aeruginosa might be the quality of LptA/LptD–thanatin interaction.

KRAS is one of the most widely prevalent proto-oncogenes in human cancers. The constitutively active KRAS oncoprotein contributes to both tumor onset and cancer development by promoting cell proliferation and anchorage-independent growth in a MAPK pathway-dependent manner. The expression of microRNAs (miRNAs) and the KRAS oncogene are known to be dysregulated in various cancers, while long noncoding RNAs (lncRNAs) can act as regulators of the miRNAs targetingKRASoncogene in different cancers and have gradually become a focus of research in recent years. In this review article, we summarize recent advances in the research on lncRNAs that have sponging effects on KRAS-targeting miRNAs as crucial mediators of KRAS expression in different cell types and organs. A deeper understanding of lncRNA function in KRAS-driven cancers is of major fundamental importance and will provide a valuable clinical tool for the diagnosis, prognosis, and eventual treatment of cancers.

Curcumin can be used for diabetes treatment. However, it rapidly degrades and has low absorption in gastrointestinal system. Its nanomicelle form might improve its efficiency. Therefore, they both assessed in streptozotocin-induced diabetic rats. Fifty male wistar rats were induced diabetes and divided into 5 groups and treated with 1) no dietary supplement, 2 and 3) 40 and 80 mg/kg curcumin, 4 and 5) 40 and 80 mg/kg nanomicelle curcumin. A group of 10 untreated rats was considered as healthy control. Serum concentrations of AST, ALT, glucose, insulin, insulin resistance, triglycerides, cholesterol, HDL-C, LDL-C and adiponectin were assessed. Body weight and weights of liver, heart and pancreas were also evaluated. Induction of diabetes increased the serum concentrations of AST, ALT, glucose, triglycerides, cholesterol, LDL-C and insulin resistance and decreased the serum concentrations of insulin, adiponectin, and HDL-C, and also body weight and weights of heart and pancreas (P

Three hundred and sixty 1-day-old male broiler chicks were randomly allocated to 4 treatments of 6 replicates to evaluate the effects of cLFchimera, a recombinant antimicrobial peptide (AMP), on gut health attributes of broiler chickens under necrotic enteritis (NE) challenge. Treatments were as follows: (T1) unchallenged group fed with corn-soybean meal (CSM) without NE challenge and additives (NC); (T2) group fed with CSM and challenged with NE without any additives (PC); (T3) PC group supplemented with 20 mg cLFchimera/kg diet (AMP); (T4) PC group supplemented with 45 mg antibiotic (bacitracin methylene disalicylate)/kg diet (antibiotic). Birds were sampled for villi morphology, ileal microbiota, and jejunal gene expression of cytokines, tight junctions proteins, and mucin. Results showed that AMP ameliorated NE-related intestinal lesions, reduced mortality, and rehabilitated jejunal villi morphology in NE challenged birds. While the antibiotic non-selectively reduced the count of bacteria, AMP restored microflora balance in the ileum of challenged birds. cLFchimera regulated the expression of cytokines, junctional proteins, and mucin transcripts in the jejunum of NE challenged birds. In conclusion, cLFchimera can be a reliable candidate to substitute growth promoter antibiotics, while more research is required to unveil the exact mode of action of this synthetic peptide.

The authors have requested that this preprint be removed from Research Square.

Imprinted genes display biased expression of paternal and maternal alleles in mammals. They are marked through epigenetic process during gametogenesis. Characterization of imprinted genes has expanded our understanding of the regulation and function of genes. In the current study, 22 experimentally validated imprinted genes in bovine (Bos Taurus) were analysed. Several supervised machine learning algorithms and attribute weighting methods were used to find characteristics of different types of imprinted genes and suggest a classification method for finding maternally and paternally expressed genes in bovine. For assessing the best model and comparing attributes in other organisms, we have also conducted a comparative analysis for human and sheep imprinted genes. According to the results of the present study, GC contents 10 and 100 kb upstream, Gly and Gln amino acids, Ile/ATC codon usage, LINE and SINE in 100kbup and length of first intron were significantly different between the maternal and paternal genes in cattle. Considering all species together, we found that GC content 100 kb up, LINE 100 kb up and the frequency of amino acids like Gly, Gln and Met were the most important attributes for identifying the paternal and maternal imprinted genes. These findings could imply conservation pattern in the attributes among these species.

This study was conducted to assess the effect of whey powder and physical form of feed on humoral immunity and transcript abundances of interleukin-4(IL-4) and interferon-gamma (In-γ) genes on 240 Ross-308 day-old-chickens. The experiment was conducted in a completely randomized design in 6 treatments in factorial arrangement (2×3), whey powder at three levels (0, 4,8%) and physical form in two levels (mash, pellet) in broiler chicken diet with 4 repeats and 10 chickens per replicate. The diets formulated based on nutritional requirements of Ross strain-308 (2009) and were fed to the birds in three periods. To determine the immunity titer of ND, IBV, IBD and influenza blood samples were collected at day 10, 31 and 42. At the end of periods, one chicken was selected from experimental units and slaughtered; lymphatic organs were weighted and gene expression of IL-4 and In-γ measured after mRNA extraction and cDNA synthesis. It was found that the addition of whey powder did not have a significant effect on the weights of the spleen and bursa of Fabricius, but the pelleting in the growing stage had a significant effect on the weight of spleen (p

Nowadays, engineered mammalian cell lines such as CHO and HEK-293 are extensively used to express recombinant proteins which require post translation modifications. Post translation modifications completely depend on presence of an appropriate signal peptide in nascent proteins. Erythropoietin is a complex protein with post translation modifications that is widely used to treat anemia, hence production this protein as a recombinant protein needs an appropriate signal peptide. The objective of this study was prediction of the best signal peptide to express recombinant erythropoietin in eukaryote system. In order to predict the best signal peptide, first, 28 signal sequences were extracted from database, then, regions and signal peptide probability of them were evaluated by Signal P version 4.1. To investigate physical and chemical features of signal sequences such as isoelectric point, net positive charge, aliphatic index and amino acid length, Protparam was applied. The results revealed that among 28 signal sequences, only 15 of them can be applied as signal peptide to express erythropoietin. Eventually, among 15 signal peptides, SCGB1D1, APOB, ALB, ULBP2 and scrg1 (respectively) were theoretically introduced as the best signal peptides to employ.

Probiotics are beneficial microorganisms and have long been used in food production as well as health promotion products. Bioengineered probiotics are used to express and transfer native or recombinant molecules to the mucosal surface of the digestive tract to improve feed efficiency and promote health. Lactococcus lactis is a potential probiotic candidate to produce useful biological proteins. The aim of this investigation was to develop a recombinant Lactococcus lactis with the potential of producing phytase. To enhance the efficiency of expression and secretion of recombinant phytase, usp45 signal peptide was added to the expression vector containing phytase gene (appA2) derived from Escherichia coli. Sequencing of recombinant plasmid containing appA2 showed the correct construction of plasmid. Total length of the phytase insert was 1.25 kbp. A Blast search of the cloned fragment showed 99% similarity to the reported E. coli phytase sequence in the GenBank (accession number: AM946981.2). A plasmid containing usp45 and appA2 electrotransferred into Lactococcus lactis. Zymogram with polyacrylamide gel revealed that the protein extract from the supernatant and the cell pellet of recombinant bacteria had phytase activity. Enzyme activity of 4 U/ml was obtained in cell extracts, and supernatant maximal phytase activity was 19 U/ml. The recombinant L. lactis was supplemented in broiler chicken feed and showed the increase of apparent digestibility on phytate phosphorus in the digestive tract and it was same as performance of E. coli commercial phytase.

Background and Aim: Short tandem repeat (STR) markers, are conserved region in human genome and highly polymorphic between individuals. Nowadays, genotyping of STR marker is widely known and used for the genetic identification of individuals in forensic DNA analyses. Based on allelic frequencies of STR loci varies between populations, investigation of genetically and forensically parameters in each population and characterization of these markers is necessary. The objective of this study was to optimizing laboratory method for application of 10autosomal STR loci (TPOX، vWA، D7S820، D8S1179، D13S317، D16S539، D18S51، D5S818، THO1، D21S311) in Kurd and Arab ethnics of Iran and investigation of population and forensic genetics parameter of these markers in these populations. Materials and Methods: In this semi-experimental study, blood samples from 93 Arab and 94 Kurd individuals were collected. After DNA extraction, PCR amplification was carried out for 10 autosomal STR loci, individually. Then, acrylamide gel electrophoresis was used to determine the genotype of each individual in each site. Results: Deviation from Hardy Weinberg equilibrium even after Bonfferroni correction was seen in the locus of D13S317in both Populations. After D13S317 loci, the highest observed heterozygosity was seen in D21S311loci for Kurd population (94%) and in THO1, vWA and D5S818 locifor Arab population (84%). The lowest observed heterozygosity (0.71 and 0.72) was seen in TPOX loci for both populations form Arab and Kurd ethnics, respectively. Investigation of forensic genetic parameters (PI, PE, PD, and PIC) showed that in except of the D13S317 loci other remaining evaluated locus had proper properties for using in genetics fingertips in both of Kourd and Arab ethnics. Conclusion: The results of current study indicate that the necessity investigation of forensic genetics for rapid characterization of the different ethnicities which located in different geographic parts of Iran in order to choose the appropriate data set to calculate of forensic genetics parameters not only within each ethnic but also between them. &nbsp

Identification of the genetic features of Persian mugger crocodile, the Gando, is crucial in management planning for conservation and preservation of this endangered species. Therefore, this study is aimed to investigate the genetic characteristics of Gando in Iran, based on population studies of the conserved and variable regions of the mitochondrial DNA. For this purpose, 12 samples of skin tissue and blood were collected from three regions of Chabahar city, south east of Iran. After DNA extraction, conserved and variable regions of the mitochondrial genome Cyt b and D-loop were amplified using PCR. The PCR products were sequenced and results were analyzed using bioinformatics software based on nucleotide data as well as protein codons. Results revealed that all samples had similar nucleotide sequences and no genetic variation was found among the samples. The closest species to Gando were Crocodylus palustris and then C. porosus. Comparison of sequences with other Crocodile species revealed the presence of 12 polymorphic loci among 1156 loci studied and also 5 haplogroups were introduced. As we didn’t find any nucleotide diversity among studied samples, therefore, to increase the genetic diversity of the Gando, it is essential to include specific conservational plans for this endangered species.

The use of anticancer peptides (ACPs) as an alternative/complementary strategy to conventional chemotherapy treatments has been shown to decrease drug resistance and/or severe side effects. However, the efficacy of the positively-charged ACP is inhibited by elevated levels of negatively-charged cell-surface components which trap the peptides and prevent their contact with the cell membrane. Consequently, this decreases ACP-mediated membrane pore formation and cell lysis. Negatively-charged heparan sulphate (HS) and chondroitin sulphate (CS) have been shown to inhibit the cytotoxic effect of ACPs. In this study, we propose a strategy to promote the broad utilization of ACPs. In this context, we developed a drug repositioning pipeline to analyse transcriptomics data generated for four different cancer cell lines (A549, HEPG2, HT29, and MCF7) treated with hundreds of drugs in the LINCS L1000 project. Based on previous studies identifying genes modulating levels of the glycosaminoglycans (GAGs) HS and CS at the cell surface, our analysis aimed at identifying drugs inhibiting genes correlated with high HS and CS levels. As a result, we identified six chemicals as likely repositionable drugs with the potential to enhance the performance of ACPs. The codes in R and Python programming languages are publicly available in https://github.com/ElyasMo/ACPs_HS_HSPGs_CS. As a conclusion, these six drugs are highlighted as excellent targets for synergistic studies with ACPs aimed at lowering the costs associated with ACP-treatment. This article is protected by copyright. All rights reserved.