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Background: Immunological factors are important in pregnancy loss because of the interaction between mother and fetus. T-regulatory cells as the component of humeral immune response play important role in the fetu-maternal interface. One of the regulatory mechanisms for these cells is mediated by antigen independent co-stimulatory signals and interaction of Cytotoxic T-Lymphocyte Antigen 4 (B7/CTLA-4) is one of these signals. The CTLA-4 which down regulates the activation and proliferation of T-cells occurs in a competitive interaction with CD28 to bind to B7. The aim of this study was to find out the relationship of CTLA-4 +49A/G gene with Recurrent Miscarriage in a group of Iranian women. Methods: In the present study, 60 women with the history of two or more pregnancy loss were selected and considered as the case group. A group of women (n=60) with at least two live births without any previous history of pregnancy loss and autoimmune diseases were taken as control group. Genomic DNA was extracted from whole blood using standard protocols. The CTLA-4 +49 A/G were detected using polymerase chain reaction-restriction fragment length polymorphisms assay. Results: The results showed that CTLA-4 +49 A/G polymorphisms were not significantly different in women with the history of two or more pregnancy loss compared to normal individuals. The frequency of G-allele polymorphism was 39.16% and 35.83% in patients and controls respectively. Conclusions: The data presented may suggest that the CTLA-4 is not associated with recurrent miscarriage in an Iranian population in Northwest region.

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In this study, 100 fish (Astronotusocellatus) with density of 8 to 12 fish per aquarium distributed to four treatments (Control, 0/25%, 0/5% and %1 Macrogard) the experiment lasted forsix weeks. In this experiment it was observed that the number of white blood cells, especially neutrophils and serum the lysozyme level in fish fed with different levels Macrogard were significantly increased compared to the control group (p<0/05). Most changes in the white blood cell count was observed in fourth week of feeding for all Macrogard levels. This material affects the immune system of Astronotusocellatus, and we can say that the all levels Macrogard were safe. They are most effective when the fish are fed for 4 weeks with diets containing Macrogard.

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The effect of Pediococcus pentosaceus as a probiotic on the growth and immunological factors of the Siberian sturgeon, Acipencer baerii, was assessed. For this purpose, 180 fish specimens of 143.3±0.01 g body weight were randomly distributed in 12 fiberglass  tanks (15 fish per tank) and fed with four diets containing 10⁷, 10⁸ and 10⁹  bacterial cells per gram, respectively. The forth diet (control) contained no probiotics. Results showed significant improvement in FCR, PER, SGR and BWI in groups fed on diets containing probiotics compared to the control group. However, CF decreased in the probiotic- treated groups, though not significant. The result of immunological factors indicated increased lyzozyme activity, ACH50 and total igM. The best result from the assessment of immune factors was observed in the fish fed with 10⁹ bacteial cell per gram of diet.

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Aims: Methicillin-resistant Staphylococcus aureus (MRSA) is recognized as an important health problem worldwide. To counteract the human innate immunity, S. aureus produces a number of immune evasion cluster (IEC) including staphylokinase (SAK), staphylococcal enterotoxin P (SEP), staphylococcal enterotoxin A (SEA), staphylococcal complement inhibitor (SCIN), and chemotaxis inhibitory protein (CHIP) encoded by sak, sep, sea, scn, and chp genes, respectively. These genes are carried by β-hamolysin-converting bacteriophages. The present study was conducted to determine the IEC phage types and antibiotic resistance patterns in 145 clinical MRSA isolates from Khuzestan Province, Iran.
Methods: All the isolates were investigated by disc diffusion method and PCR assay of sak, sep, sea, scn, and chp genes.
Findings: The assessment of antibiotic resistance showed the highest rate of resistance towards penicillin (97.25%), followed by methicillin (95.8%), ceftazidime (81.4%), erythromycin (71.8%), clindamycin (61.4%), ciprofloxacin (60.7%), gentamycin (56%), imipenem (56.55%), and vancomycin (0%), respectively. Also, the frequency of IEC types was as follows: type A (4.8%), type B (9%), type C (13.1%), type D (12.4%), type E (27.6%), type F (1.4%), type G (0.7%), and type H (6.9%). On the other hand, 24.1% of the isolates did not show any of the IEC types.
Conclusion: The findings showed that IEC-carrying bacteriophages are highly prevalent among MRSA strains, resulting in the adaptation and counteraction of bacteria to the human immune system. Therefore, understanding the role of IEC in the virulence of bacteria can improve our knowledge about the evolution, vaccination, and treatment of S. aureus infection.

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The effect of human chorionic gonadotropin (hCG) on the immune response, biochemical parameters and serum enzymes in adult male and female goldfish during breeding season was investigated. Mature male and female received an intramuscular injection of human chorionic gonadotropin (hCG) 1/ IU hCG g/ BW and the control group was injected with saline solution. Twelve and 24 hours after hCG injection, blood samples were taken from broodstock and immune parameters (IgM, IgG, C3, C4), biochemical and enzymes (glucose, uric acid, urea, cholesterol, total protein, triglycerides, calcium, creatinine, albumin, HDL, LDL, AST and ALT) were measured. There were no significant changes in IgM levels in females between 12 and 24 hours after hCG injection compared to the control group (P>0.05). However, IgM level in male showed a gradual decrease in comparison to control fish (P<0.05). On the other hand, the concentration of IgM and C4 complement decreased by passing time after hCG injection in both male and female (P<0.05). The C3 complement dramatically increased after hCG injection in both male and female (P<0.05). Injection of human chorionic gonadotropin caused a significant change in serum biochemical and enzyme in both male and female. The results showed that the hCG hormone even in a single dose within a short time after injection, strongly stimulate the immune system and changes physiological condition of adult male and female goldfish.

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Background: Infectious agents are considered as one of the possible etiological factors of systemic lupus erythematosus (SLE). It has been suggested that torque teno virus (TTV) may trigger autoimmune disorders, but few studies have been conducted on the relationship between this virus and autoimmune diseases, especially SLE. The present study aimed to evaluate the association between TTV frequency and SLE.
Materials & Methods: Serum samples were collected from a total of 116 participants, including 58 healthy people and 58 SLE patients who referred to the rheumatology clinic of Shahid-Beheshti hospital in Kashan, Iran from January 2020 to January 2021. After the extraction of viral DNA from the samples, a nested PCR test was performed using specific primers to detect TTV.
Findings: TTV was detected in 43 SLE patients (74.1%, 95% CI: 63.4-86.2) and 33 healthy individuals (56.9%, 95% CI: 44.1-69.0). A significant correlation was found between SLE and the presence of TTV (r= .32, p= .03). There was no correlation between the presence of TTV and musculoskeletal involvements, skin lesions, renal manifestations, and hematological manifestations (r< .05, p> .05). TTV was detected more frequently in patients with active lupus than in patients with quiescent disease, and this difference was significant (p= .048).
Conclusion: A significant association between TTV and SLE was observed in the present study; however, further studies are needed to investigate the role of TTV in the pathogenesis and clinical course of SLE.

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Because of the importance of osmoregulation in success of the juvenile Caspian Roach (Rutilus caspicus) release, maintenance and their restoration, this study was performed with the aim of the evaluation of these fish (average weight of 1.4 g± 0.018 g) to the salinity tolerance of the Caspian Sea by abrupt and gradual transfer methods on two different occasions of short-term (24 hours) and long-term (21 days). For this reason, 360 juvenile roaches were distributed to 9 different experimental treatments (each of them with 3 replicates) with 1) increased abrupt salinity (direct transfer to the Caspian Sea water), 2) increased gradual salinity (3 ppt for every 24 hrs) and 3) the Control group (freshwater). In order to evaluate the ion regulation, osmolality, Na⁺, K⁺ and Cl^- were measured in fish. Some immune responses related to the increased salinity (immunoglobulin and lysozyme) and stress indices (cortisol and glucose) were also measured. Based on the obtained results, the fish in both treatments of increased salinity showed high percentage of survival. All parameters related to ion regulation were decreased after a primary increase. Furthermore, the trend of cortisol and immunoglobulin changes were negatively correlated showing decreased stress response and increased immune system for both salinity transfer methods. Thus, the roach juveniles (1-2 g) are able to be transferred into the Caspian Sea directly.

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This experiment evaluated the effect of soluble polysaccharides in Spirulina platensis microalgae on growth performance, body composition and immune response of rainbow trout (Oncorhynchus mykiss). In this regard, 180 rainbow trout (17.22 ± 0.5g) were selected and distributed in five experimental treatments (with three replications). Treatments included different levels of polysaccharide in dosage of 0 (control), 500, 1000, 2000 and 3000 mg / kg of feed. According to the results, in growth factors (final weight, body weight gain, specific growth factor, protein efficiency, feed conversion ratio and condition factor), no significant difference was observed. The highest and the lowest amount of carcass proteins were observed in the diet containing 500 mg polysaccharide and control, which showed a significant difference. The amount of fatty acids of the muscle of the fish body did not show any significant difference among the treatments. The highest and lowest levels of lysozyme activity were observed in treatments of 2,000 mg polysaccharide and control and the highest and lowest levels of complement activity were observed in treatments containing 3000 mg polysaccharide and control respectively (P <0.05). In general, the use of polysaccharide extracted from micro-algae did not significantly improve rainbow trout growth, but a slight improvement in growth and body composition (protein) in 500 mg polysaccharide per kg of diet was observed. In terms of safety indicators, treatments of 2,000 and 3,000 mg of polysaccharides per kg of diet had a good performance and could be used whenever needed.
 

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The cotton bollworm, Helicoverpa armigera (Hübner), is one of the most important pests of crops. Like other insects, this insect has an immune system against external threats such as various diseases, parasitoids, and chemical compounds. Phenoloxidase is a key factor in the immunity of insects and other arthropods. The present study investigated the lethal effects of four insecticides, indoxacarb, profenofos, chlorfluazuron, and hexaflumuron, and their effects on phenolic oxidase activity in cotton bollworm. Among the studied insecticides, in two bioassays (mixture of insecticide with artificial food and immersion of sugar beet leaves), Chlorfluazuron had the highest toxicity (LC₅₀ = 1.71 and 3.11 mg ai/liter, respectively). Also, the highest phenol-oxidase activity at 24 h was for larvae treated with chlorfluazuron and hexaflumuron. Also, when treated with chlorfluazuron and hexaflumuron, the larvae fed on an artificial diet had higher phenoloxidase activity than those fed on sugar beet cultivars.

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A research study was conducted to evaluate the effects of three herbal extracts and an antibiotic –virginiamycin- on growth performance, immune system, blood factors and selected intestinal bacterial populations in broiler chickens. A total of four hundred and eighty 1-day old male broiler chicks were assigned to the basal diet (control) , basal diet supplemented with 15 ppm of virginiamycin, basal diets with a 0.1% dose of either thyme (Thymus vulgaris), coneflower (Echinacea purpurea), garlic (Allium sativum) or a blend of the three extracts in the drinking water. The highest and the lowest body weight and weight gain were related to virginiamycin and coneflower, (P< 0.05) respectively. The lowest and the highest feed conversion rates were respectively related to virginiamycin and coneflower (P< 0.05). Relative weight of bursa Fabricius in the garlic group showed a significantly more increase as compared with other groups, while the relative weight of spleen was unaffected by treatments. Cutaneous basophils hypersensitivity response (to phytohemaglutinin injection) and antibody response to Sheep Red Blood Cells (SRBC) was higher in coneflower group (P< 0.05). Antibody responses to Newcastle Disease vaccine (LaSota) was unaffected by treatments but coneflower improved antibody levels (P> 0.05). Garlic (Allium sativum) significantly reduced the serum levels of cholesterol, LDL, and triglyceride as well as significantly increasing the level of HDL. Thyme (Thymus vulgaris) improved hematocrit percentage and hemoglobin concentration, but not significantly. The colony forming units of Escherichia coli in digesta of ileo-cecum in the blend group showed a significantly lower number compared with control. However, there was no difference observed in E. coli counts between blend group and others, except for control. The lactic acid bacteria counts in the thyme group increased as compared to other groups, except for coneflower (P< 0.05).

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In the last month of 2019, an unknown virus appeared in Wuhan, China. Sequencing studies have shown that the virus is a new member of the coronavirus family, which mostly causes a respiratory disease with pneumonia-like symptoms. The new coronavirus consists of 25 proteins, including 4 important structural proteins and 15 non-structural proteins. Spike protein is one of the most important structural proteins on the surface of the virus; It is highly glycosylated and plays a key role in the virus binding to the host cells. The binding of glycans to proteins affects their structure and function in two ways; They lead to proper protein folding, and can play an important role in protein interactions, and also, by covering the surface of the protein, it causes the virus to escape from the immune system. So it is obvious that the study of glycan structures becomes more important when either a vaccine is going to be designed or glycan structures have important roles in the folding, activity, and interaction of a protein. Therefore, since the spike protein is a non-functional structural protein, the study of glycan structures is important for two goals of vaccine design and investigating the role of glycans in protein interactions. In this article, we are going to review the most important findings on spike protein glycosylation and compare the amount of glycosylation in different viruses, indicating the importance of glycan structures in designing an effective vaccine.

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This study was conducted to investigate the effects of the medicinal plant yarrow (Achillea millefolium L.), a probiotic (Primalac) and an antibiotic (virginiamycin) on gastrointestinal tract (GIT) characteristics, microbial populations, immune response, serum lipids and growth performance of broiler chickens. A total of 250 one-day old male broilers (Ross 308) were randomly allocated to 5 treatments, 5 replicates with 10 birds in each in a completely randomized design. Experimental treatments included the control, yarrow powder at two different concentrations (1.5 and 3% of diet), Primalac (0.1% of diet) and virginiamycin (15 ppm). The highest feed conversion ratio (FCR) was observed in the control while the lowest FCR was seen in the virginiamycin group at 42 days of age (P< 0.05). Moreover, the highest body weight gain (BWG) was observed in the virginiamycin group while the lowest value was related to the control animals (P< 0.05). Carcass yields were not different between treatments (P> 0.05). Relative weights of breast and thigh were similar among all treatments (P> 0.05). Relative weights of bursa Fabricius, spleen and primary immune response (total titer, IgG and IgM) against sheep red blood cells (SRBC (were not affected by treatments. The serum cholesterol, triglyceride as well as high and low density lipoprotein (LDL and HDL) levels were different among treatments (P< 0.05). The lowest concentrations of the mentioned parameters were obtained in the group supplemented with 3% of yarrow (P< 0.05). The highest and lowest antibody titers (secondary immune response) against SRBC were observed by the yarrow (3%) and antibiotic supplementations, respectively (P< 0.05). The highest lactic acid bacteria (LAB) counts were detected in the crop, ileum and cecum of the Primalac group (P< 0.05). Inclusion of virginiamycin and yarrow (3%) caused a significant decrease in coliforms and total aerobic bacteria counts in crop, ileum and cecum (P< 0.05). The results of this study showed that the administration of yarrow (3%) can reduce the levels of serum lipids and boost the immune response in broilers. Moreover, it led to reduced pathogenic bacteria population in the GIT which could help to improve intestinal health and well being of poultry. It is proposed that yarrow can be used as an antibiotic alternative.

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Objective: This study investigated the effects of adipose-derived mesenchymal stem cells (MSCs) and conditioned medium injection on cell infiltration in the brains of an experimental C57BL/6 mice model of autoimmune encephalomyelitis (EAE). Methods: EAE was induced with myelin oligodendrocyte glycoprotein (35-55 peptides) in 20 mice. MSCs were obtained from the adipose tissue of C57BL/6 mice and cultured with Eagle’s minimum essential medium/alpha medium (DMEM) after removal of non-adherent cells. After 2 to 3 passages, 15 days after induction of EAE, 4 groups (n=5) of C57BL/6 mice were treated as follows: i) MSCs were injected intraperitoneally, ii) MSCs were injected intravenously, iii) conditioned medium was administrated intraperitoneally, and iv) the control group received no treatment. After day 60, the mice brains were removed and the effect of adipose-tissue MSCs and administration of conditioned medium was investigated. Results: Leukocyte infiltration and clinical scores were significantly reduced in animals that received MSC and conditioned medium compared to untreated animals. Body weight increased significantly in the treated groups compared to the control group. Percentage of survival also increased in the animals that received MSCs and conditioned medium as compared to the control group. Conclusion: MSCs had immunomodulatory and neurogenerative functions which reduced leukocyte infiltration and improved clinical scores in the EAE animals that received MSC and conditioned medium compared to untreated animals with EAE.

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Immune checkpoints are molecules that regulators  the immune system. However, some tumor cells can express the ligands of immune checkpoints to escape from antitumor immune responses. Some agents, such as antibodies, can inhibit these checkpoints that prevent the immune system from targeting and killing cancer cells. The aim of this study was to express a novel bispecific diabody in periplasmic space of E.coli for simultaneous targeting of two immune checkpoints, cytotoxic T‑lymphocyte‑associated protein 4 (CTLA‑4) and programmed death- ligand 1 (PD‑L1).
The bispecific diabody was constructed based on the variable regions gene of anti PD-L1 and anti CTLA‑4 antibodies. The optimum codon for expression in E. coli was chemically synthesized and subcloned in pET21 expression plasmid. After transformation, the effect of cultivation conditions on periplasmic expression of the protein in E. coli BL21(DE3) was evaluated. Then, the bispecific diabody was purified .
Expression of diabody with a molecular weight of 55 kDa was verified by Sodium dodecyl sulfate‑polyacrylamide gel electrophoresis and western blotting analysis. The best condition for soluble periplasmic expression was obtained to be incubation with 0.5 mM isopropyl β‑D‑1‑thiogalactopyranoside at 23°C. The protein was successfully purified using affinity chromatography with a final yield of 0.4 mg/L. The affinity of the purified protein interaction were checked by ELISA.
Recombinant Diabody protein was cloned, expressed, and purified in a bacterial system and Diabody Interaction with PDL-1 receptor conformed by Cell-Elisa.

 

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In the present study, simultaneous evaluation for the survival ability of probiotic bacteria at pH 2.5, viability in the presence of pepsin-pancreatin, resistance to bile salt, Adhesion ability, bacterial resistance against antibiotic substances with measuring of minimum inhibitory concentration, auto-aggregation and co-aggregation with interference of (Escherichia coli (NCTC 12900 O157:H7), Salmonella enterica (ATCC 13076) and Listeria monocytogenes (ATTC 7644)) were investigated. In the test of Acid tolerance, it was found that all the strains except LF 55, 61 and F had high viability. Strains LF 56, LF 57, LF 55, O, F and strains LF 56, LF 57 and A7 showed that had the most resistant against pancreatin and in the presence of pepsin, respectively. Furthermore, all of the strains except La5, 52 and 49 were highly resistant to bile salts. Among the 19 strains, Lactobacillus gasseri had the highest adhesion ability (62.90%). Strain Lactobacillus plantarum LF 56 and Lactobacillus gasseri 54 C showed that had the highest potentials for auto-aggregation and co-aggregation (75.31% and 33%) on three pathogens, respectively. Several strains of Lactobacillus plantarum were sensitive to the studied antibiotic substance. However, strains M8 and M11 were resistant to Kanamycin and Chloramphenicol, and also, LF 55 and D showed high resistance against Ampicillin and Chloramphenicol, respectively. It was, moreover, found that Kanamycin, Tetracycline and Chloramphenicol had no significant influence on strains 61. It was demonstrated that Lactobacillus gasseri had medium resistance to Ampicillin, Erythromycin and Tetracycline, while Lactobacillus fermentum showed highly resistance to Ampicillin and Erythromycin. According to the results, human-isolated Lactobacillus gasseri gained the high scores in terms of probiotic properties as compared with other strains. Therefore, Lactobacillus gasseri can potentially protect epithelium tissue of intestine against pathogenic bacteria and it is a preferred candidate for prophylactic purposes, preventing of intestine infections and increasing of body immune system.
 

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Aims: New vaccines based on recombinant and DNA proteins are safer than traditional vaccines, but unfortunately, they have lower Therefore, there is a need for the development of safe and strong that can increase the immune PLGA), ester, consists of acidic and lactic acid. Its hydrolysis leads to the production of lactic acid and glycolic acid monomers. The aim of this study was to compare humoral and cell mediated immune response to coated PLGA in mice.
Materials and Methods: In this experimental study, PLGA nanoparticles were produced by water/oil (W/O) method. Tetanus toxin attached to by EDC. After coated characterization, they were injected into different groups of mice. The complete and Alum as After a single injection, the of was investigated by ELISA and cellular analyzed by spleen cell proliferation assay. One-way analysis of variance was used.
Findings: PLGA nanoparticles had a strong effect, and when used with antigens, could produce cellular and humoral immune response far more powerful than alum and than Freund’s adjuvant.
Conclusion: Glycolic polyester, in the form of conjugation with an antigen, can be used to increase the immune response, especially in the cellular immune arm, relative to the antigenic solution. Although PLGA seems not so successful to the humoral immune stimulus against in comparison to the full of it can be a significant competitor with

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The purpose of this study was to evaluate the effect of sumac (Rhus coriaria L.) administration on the growth performance, immune response, and haematological parameters of rainbow trout (Oncorhynchus mykiss). Rainbow trout juveniles with an initial weight of 38±2.65 g were allocated into 12 tanks at a density of 15 fish per tank. Fish were fed four experimental diets containing 0% (as control diet), 0.5%, 2%, and 5% sumac-supplementation for 56 days. After the feeding trial, fish were challenged with Yersinia ruckeri, and survival rate was calculated for 15 days. Sumac diet significantly increased resistance to the pathogen and led to the control of infection in rainbow trout without changing weight. The leukocyte (WBC) and erythrocyte (RBC), lymphocyte, monocyte, and neutrophil value was significantly higher in fish that were fed a sumac-supplemented diet when compared with the control. Serum lysozyme, and alternative complement pathway haemolytic activity (ACH50) as well as the hepatic expression of TNF-α and IL-1b were recorded to be the highest in fish fed 2% and 5% sumac-supplement. Meanwhile, mRNA expression levels of IL-10 significantly decreased in fish fed 5% sumac supplementation for 56 days. These results suggest that sumac administration, especially 2% and 5%, may effectively enhance the immune system, resistance to the pathogen, and hematopoiesis in rainbow trout.