Effect of IL-2 co-expressed or co-inoculated with immuno-dominant epitopes from VP1 protein of FMD virus on immune responses in BALB/c mice

Objective(s): The results of studies on vaccine development for foot-and-mouth disease (FMD) virus show that the use of inactivated vaccines for FMD virus is not completely effective. Novel vaccinations based on immuno-dominant epitopes have been shown to induce immune responses. Furthermore, for safety of immunization, access to efficient adjuvants against FMD virus seems to be critical.Materials and Methods: In this study, we produced epitope recombinant vaccines from the VP1 protein of the FMD virus for serotype O of Iran. Constructs were included polytope (tandem-repeat multiple-epitope), polytope coupled with interleukin-2 (polytope-IL 2) as a molecular adjuvant and IL-2. Three expression vectors were constructed and expressed in Escherichia coli BL21 (DE3). To evaluate whether these recombinant vaccines induce immune responses, BALB/c mice were injected with the recombinant vaccines and their immune responses were compared with a negative control group. The humoral and cellular immune responses were measured by ELISA.Results: The results showed that IL-2 co-expressed or co-inoculated with Polytope protein enhances the immune effect of multiple epitope recombinant vaccine against FMD virus. The results of total immunoglobulin G (IgG), IgG1, and IgG2a levels and secretion of interferon gamma (IFN-γ), IL-4 and IL-10 revealed that there were significant differences between negative control group and other injected mice with the recombinant vaccines (

Comparative Analysis of Peripheral Alkaline Phytase Protein Structures Expressed in E. coli

Background: Degradation of phytic acid to inorganic phosphate in domestic animals’ diets requires thermostable phytase. Although Basillus subtilis phytase shows a potential to be degraded phytate complex in high temperature, the enzyme activities and yields need to be increased to make them possible for industrial application. Methods: The phytase gene from Bacillus subtilis DR8886 was isolated from Dig Rostam hot mineral spring in Iran and cloned into pET21(+) and pET32(+). Expression was induced with 1.5 mM IPTG and the proteins were purified. Results: The recombinant protein affected by thioredoxin (Trx) from pET32a-PhyC was estimated to constitute about 31% of the total soluble protein in the cells; its concentration was 3.5 μg/ml, and its maximal phytase activity was 15.9 U/ml, whereas the recombinant phytase from pET21a-PhyC was estimated to comprise about 19% of the total soluble protein; its concentration was 2.2 μg/ml, and its maximal phytase activity was 69 U/ml. The molecular masses of recombinant phytase with and without Trx were about 60 kDa and 42 kDa, respectively. Zymography confirmed that the recombinant enzymes were active. Although the concentration of the alkaline phytase expressed by pET32a was approximately 59% greater than that expressed by pET21, its phytase activity was approximately 77% less. Conclusion: This study showed that the peripheral gene (Trx) encoded by the pET32a (+) vector are the principal reason for the decrease in recombinant phytase enzyme activity

Quercetin in Food: Possible Mechanisms of Its Effect on Memory

Abstract: Quercetin (3,3�,4�,5,7-pentahydroxyflavone) is found in vegetables and fruits. It is one of the major flavonoids that is part of human diets. Quercetin has several pharmacological effects in the nervous system as a neuroprotective agent. In this review, we summarize the research on quercetin and its role in memory in both animals and humans. Articles were chosen from the Scopus, PubMed, and Web of Science databases. In this review, we describe and summarize the importance of quercetin’s presence in the body, particularly in the brain; its kinetics, including its absorption, metabolism, distribution, and excretion; its behavioral effects; and some of the possible mechanisms of action of quercetin on memory in different animal models. Several important pathways that may be involved in the processes of learning and memory, long-term potentiation, and cognition may be impaired during neurological diseases or other medical conditions. As dietary quercetin is important, provision of its best formulation for delivery to the brain as a nutraceutical and in clinical translational research for the prevention or treatment of Alzheimer’s disease and other types of dementia is necessary. Keywords: Alzheimer’s disease, flavonoids, food, memory, querceti

Up-regulation of wheat nitric oxide synthase gene in response to Zataria multiflora essential oil dispersion

Abstract Overproduction of nitric oxide (NO) results in nitrative stress, in turn causing peroxidation of lipids, oxidation of protein and damage to nucleic acids among many others. Presumbly, a simple mode to suppression of nitrative stress is found to be a natural antioxidant from aromatic plants. Zataria multiflora essential oil (ZO) is known for its potent antioxidant activities. Highly insoluble in water, ZO can be easily degraded by light, oxygen, high temperature and extreme pH. Dispersion of ZO into polyvinylpyrrolidone (PVP) might enhance its stability and self-life while storage and applications. The present research assesses in vitro antioxidant function of PVP/ZO dispersion versus nitrite and nitric oxide (NO). Followed by this efficiency of the PVP/ZO dispersion on the production of nitric oxide synthase (NOS) mRNAs in the wheat seedlings in hydroponic condition was evaluated. PVP and PVP/ZO dispersions characterized with negative zeta-potential was a non-Newtonian shear-thickening fluid. PVP/ZO was fraught with effective in vitro nitrite (IC 50 , 160 µg/mL) and NO (IC 50 , 168 µg/mL) scavenging activities. The result indicated that PVP/ZO dispersion up-regulated NOS (3.5 folds, at 30 µg/mL) mRNA production while down-regulated it at higher concentration. Hence, once applied at low concentration PVP/ZO can be promising as plant modulator for plant growth regulation

Estimating occupant satisfaction of HVAC system noise using quality assessment index

Noise may be defined as any unwanted sound. Sound becomes noise when it is too loud, unexpected, uncontrolled, happens at the wrong time, contains unwanted pure tones or unpleasant. In addition to being annoying, loud noise can cause hearing loss, and, depending on other factors, can affect stress level, sleep patterns and heart rate. The primary object for determining subjective estimations of loudness is to present sounds to a sample of listeners under controlled conditions. In heating, ventilation and air conditioning (HVAC) systems only the ventilation fan industry (e.g., bathroom exhaust and sidewall propeller fans) uses loudness ratings. In order to find satisfaction, percent of exposure to noise is the valuable issue for the personnel who are working in these areas. The room criterion (RC) method has been defined by ANSI standard S12.2, which is based on measured levels of in HVAC systems noise in spaces and is used primarily as a diagnostic tool. The RC method consists of a family of criteria curves and a rating procedure. RC measures background noise in the building over the frequency range of 16-4000 Hz. This rating system requires determination of the mid-frequency average level and determining the perceived balance between high-frequency (HF) sound and low-frequency (LF) sound. The arithmetic average of the sound levels in the 500, 1000 and 2000 Hz octave bands is 44.6 dB; therefore, the RC 45 curve is selected as the reference for spectrum quality evaluation. The spectral deviation factors in the LF, medium-frequency sound and HF regions are 2.9, 7.5 and -2.3, respectively, giving a Quality Assessment Index (QAI) of 9.8. This concludes the QAI is useful in estimating an occupant′s probable reaction when the system design does not produce optimum sound quality. Thus, a QAI between 5 and 10 dB represents a marginal situation in which acceptance by an occupant is questionable. However, when sound pressure levels in the 16 or 31.5 Hz octave bands exceed 65 dB, vibration in lightweight office construction is possible

Quantiation of IL-4, IL-10 and IFN- Genes Expression after Immunization of Mice with CFP-10 and ESAT-6 Containing Vectors

ABSTRACT Background: Tuberculosis is a disease with high morbidity, caused mainly by Mycobaterium tuberculosis (M.tb.). DNA vaccines show a promising future due to their unique advantages over conventional methods. The early-secreted antigen target (ESAT)-6 and culture filtrate protein (CFP)-10 of M.tb. antigens have been identified as vaccine candidates against Mycobacteria and used as subunit vaccines, DNA or protein, in different studies. Objective: To investigate the potential of pcDNA3.1+ plasmid containing CFP-10 and ESAT-6 genes in induction of local immune responses after intramuscular injection in BALB/c mice. Methods: pcDNA 3.1+ CFP-10 and pcDNA3.1+ ESAT-6 plasmids were prepared and defined groups of mice were injected intramuscularly with the plasmids both separately and in combination. The RNA was extracted from muscles after one month and cDNA was made using RT-PCR. The expressions of IL-4, IL-10 and IFN-γ genes cytokines were evaluated using comparative real time PCR. Results: Expression of IL-4 and IL-10 increased in the injection site of the mice groups which received plasmids encoding ESAT-6 and CFP-10 individually or together. More than 10-fold increase in IFN-γ expression was found in samples taken from mice groups inoculated by plasmids encoding ESAT-6 and CFP-10 individually or together. Conclusion: pcDNA 3.1+ESAT-6 and pcDNA3.1+CFP-10 plasmids can increase the expression of IFN-γ in mice after immunization

Design and Construction of Chimeric VP8-S2 Antigen for Bovine Rotavirus and Bovine Coronavirus

Purpose: Bovine Rotavirus and Bovine Coronavirus are the most important causes of diarrhea in newborn calves and in some other species such as pigs and sheep. Rotavirus VP8 subunit is the major determinant of the viral infectivity and neutralization. Spike glycoprotein of coronavirus is responsible for induction of neutralizing antibody response. Methods: In the present study, several prediction programs were used to predict B and T-cells epitopes, secondary and tertiary structures, antigenicity ability and enzymatic degradation sites. Finally, a chimeric antigen was designed using computational techniques. The chimeric VP8-S2 antigen was constructed. It was cloned and sub-cloned into pGH and pET32a(+) expression vector. The recombinant pET32a(+)-VP8-S2 vector was transferred into E.oli BL21CodonPlus (DE3) as expression host. The recombinant VP8-S2 protein was purified by Ni-NTA chromatography column. Results: The results of colony PCR, enzyme digestion and sequencing showed that the VP8-S2 chimeric antigen has been successfully cloned and sub-cloned into pGH and pET32a(+).The results showed that E.coli was able to express VP8-S2 protein appropriately. This protein was expressed by induction of IPTG at concentration of 1mM and it was confirmed by Ni–NTA column, dot-blotting analysis and SDS-PAGE electrophoresis. Conclusion: The results of this study showed that E.coli can be used as an appropriate host to produce the recombinant VP8-S2 protein. This recombinant protein may be suitable to investigate to produce immunoglobulin, recombinant vaccine and diagnostic kit in future studies after it passes biological activity tests in vivo in animal model and or other suitable procedure

Construction of Eukaryotic Expression Vectors Encoding CFP-10 and ESAT-6 Genes and Their Potential in Lymphocyte Proliferation

Background: Mycobacterium (M.) bovis is the agent of bovine tuberculosis (TB) in a range of animal species, including humans. Recent advances in immunology and the molecular biology of Mycobacterium have allowed identification of a large number of antigens with the potential for the development of a new TB vaccine. The ESAT-6 and CFP-10 proteins of M. bovis are important structural and functional proteins known to be important immunogens. Methods: In the current study, the DNAs encoding these genes were utilized in the construction of pcDNA 3.1+/ESAT-6 and pcDNA3.1+/CFP-10 plasmids. After intramuscular injection of BALB/c mice with these plasmids, ESAT-6 and CFP-10 mRNA expression was assessed by RT-PCR. Mice were inoculated and boosted with the plasmids to evaluate their effects on lymphocyte proliferation. Results: Our results indicate the plasmids are expressed at the RNA level and can induce lymphocyte proliferation. Conclusion: Further study is needed to characterize the effect of these antigens on the immune system and determine whether they are effective vaccine candidates against M. bovis

Gene expression profile analysis of residual feed intake for Isfahan native chickens using RNA-SEQ data

The factors that affect residual feed intake (RFI) of two chicken breeds (Ross as commercial and Isfahan as Iranian local breed with low and high RFI, respectively) were examined. We sequenced their liver transcriptomes using RNA-Seq while focusing on the identification of important candidate genes, which might influence RFI and growth rate. The differential gene expression analysis revealed that 121 and 279 known genes were significantly up- and down-regulated in the local breed, respectively. QTL enrichment analysis revealed that 63 down-regulated genes in the local breed were enriched in the QTL regions related to feed efficiency traits. Moreover, the functional enrichment analysis showed that the down-regulated genes in the native chickens were mainly involved in the different metabolic processes, such as carboxylic acid metabolic process and response to stress. The Differentially Expressed Genes explored between the chickens of the two breeds led to the identification of some important candidate genes for further breed improvement programmes, including RSAD2, IL15, LIPI, EGR1 and DUSP16. These findings will be a useful resource for the biological investigations of RFI-related genes in Isfahan local chickens and may provide some clues for understanding the molecular genetic mechanisms in other chicken breeds.highlights A global view of transcriptome differences in the liver tissues of commercial and local chicken breeds with different RFI values is obtained. This investigation will contribute to the improvement of chicken genome annotation through the identification of novel transcripts and novel gene. Some important candidate genes related to RFI is obtained