Enhancement of immune responses by vaccine potential of three antigens, including ROP18, MIC4, and SAG1 against acute toxoplasmosis in mice

Toxoplasma gondii (T. gondii) causes considerable financial losses in the livestock industry and can present serious threats to pregnant women, as well as immunocompromised patients. Therefore, it is required to design and produce an efficient vaccine for controlling toxoplasmosis. The present study aimed to evaluate the protective immunity induced by RMS protein (ROP18, MIC4, and SAG1) with Freund adjuvant, calcium phosphate nanoparticles (CaPNs), and chitosan nanoparticles (CNs) in BALB/c mice. The RMS protein was expressed in Escherichia coli (E. coli) and purified using a HisTrap HP column. Thereafter, cellular and humoral immunity was assessed by injecting RMS protein on days 0, 21, and 35 into four groups [RMS, RMS-chitosan nanoparticles (RMS-CNs), RMS-calcium phosphate nanoparticles (RMS-CaPNs), and RMS-Freund]. Phosphate buffered saline (PBS), CNs, CaPNs, and Freund served as the four control groups. The results displayed that vaccination with RMS protein and adjuvants significantly elicited the levels of specific IgG antibodies and cytokines against toxoplasmosis. There were high levels of total IgG, IgG2a, and IFN-γ in vaccinated mice, compared to those in the control groups, especially in the RMS-Freund, indicating a Th-1 type response. The vaccinated and control mice were challenged intraperitoneally with 1 × 103 tachyzoites of the T. gondii RH strain four weeks after the last injection, and in RMS-Freund and RMS-CaPNs groups, the highest increase in survival time was observed (15 days). The RMS can significantly increase Th1 and Th2 responses; moreover, multi-epitope vaccines with adjuvants can be a promising strategy for the production of a vaccine against toxoplasmosis

Ganoderma lucidum: A promising anti-inflammatory medicinal plant

Inflammation is a complex process and part of the host immune defense against invading micro-organism or trauma. Over production of some pro-inflammatory mediators can lead to chronic diseases of the inflammatory origin. Medicinal Plants which are used as anti-inflammatory agents, mainly act affecting various stages of the process of inflammation. In general they can inhibit formation of a wide of mediators such as cytokines by immune cells to prevent the inflammatory reaction cascade from starting. The use of most of the medicinal plants in treatment of chronic disease of the inflammatory origin is based on clinical and pharmacological trials. Meanwhile, the use of most of them is based on their longstanding traditional use in folk medicine. In this review, we report some of anti-inflammatory effects of G. lucidum as an ancient Chinese herbal medicine

The effect of ethanolic extract of Thymus kotschyanus on cancer cell growth in vitro and depression-like behavior in the mouse

Cancer and depression are known as two of the most debilitating disease and disorder increasing evidence suggest an urgent need for new therapeutic agents with lower toxicity and high efficacy. Some Thyme species extracts have remarkably been shown to positively affect depression and cancer cells. In the present study, we investigated the effect of Thymus kotschyanus on depression and cancer cells. To this end, in experiment 1, NMRI mice were treated orally with the ethanolic extract of T. kotschyanus (50, 150 and 250 mg/ml) for seven days and then depression-like behavior was measured by Forced Swim Test (FST) and Tail Suspension Test (TST). In experiment 2, the pharmacological effect of the extract on the lung (A549) and cervical (Hela) cancer cell lines was also evaluated by MTT (3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide) in various concentration_(10, 5, 2.5, 1.25, 0.63, 0.31, 0.15 and 0.08 mg/ml). The results indicated that T. kotschyanus extract treatment (150 and 250 mg/kg) decreased depression-like behavior in the FST and TST tests in adult mice. Moreover, the treatment inhibited cancer cell growth and viability in a dose and time-dependent manner. Collectively these findings suggest that T. kotschyanus have antidepressant and anticancer effects

Development and immunereactivity evaluation of a chimeric recombinant protein encoding <i>Brucella</i> antigen: <i>In silico </i> to <i>in vitro</i>

30-36Brucellosis is an important health problem in developing countries and no vaccine is available for the prevention of infection in humans. Because of clinically infectious disease and its economic consequences in human and animals, designing a proper vaccine against Brucella is desirable. In the present study, we evaluated the immune responses induced by a designed recombinant chimera protein and investigated the immunogenic potential of some immune reactive antigens of Brucella. Three immune dominant antigens of Brucella including trigger factor (TF), Omp31 and Bp26 (have been characterized as potential immunogenic and protective antigens) were fused together by EAAAK linkers to produce a chimera. Recombinant chimeric protein was synthesized, cloned and expressed in Escherichia coli BL21 (structure were designed in silico). The purification of recombinant protein was performed by using Ni-NTA agarose, and anti-His antibody was used for confirmation (Western blot). The recombinant chimeric protein could be a new potential antigen candidate for the development of a subunit vaccine against Brucella. These results demonstrate the role of the Bioinformatics in vaccine design, assisted by experimental procedures

Toll-like receptor 4 activation on human amniotic epithelial cells is a risk factor for pregnancy loss

Background: Maternal–fetal tolerance plays a fundamental role in the maintenance of pregnancy. However, this immunological tolerance can be influenced by intrauterine infections. Human amniotic epithelial cells (hAECs) have immunomodulatory effects and respond to invading pathogens through expressing various toll-like receptors (TLRs). We hypothesize that bacteria or bacterial products affect the immunosuppressive effects of hAECs through TLR stimulation. Here, we investigated how a successful pregnancy can be threatened by TLR4 activation on hAECs on lipopolysaccharide (LPS) engagement. Materials and Methods: hAECs were isolated from the amniotic membrane received from six healthy pregnant women. The immunophenotyping of hAECs was studied by flow cytometry. The isolated hAECs (4 × 105 cells/ml) were cultured in 24-well plates in the presence or absence of LPS (5 μg/ml). After 24, 48, and 72 h of incubation, the culture supernatants of hAECs were collected, and the levels of interleukin-5 (IL-5), IL-6, IL-1β, tumor necrosis factor-alpha (TNF-α), transforming growth factor-beta 1 (TGF-β1), and prostaglandin E2 (PGE2) were measured by enzyme-linked immunosorbent assay. Results: TLR4 activation showed a stimulatory effect on TGF-β1 production of hAECs (P < 0.001–0.05). PGE2 production of LPS-stimulated hAECs was significantly increased (P < 0.01–0.05). Moreover, TLR4 could induce TNF-α and IL-1β production of hAECs (P < 0.0001–0.01), while this effect was not observed on IL-6 production of hAECs. The IL-5 was produced at a very low level in two culture supernatants of hAECs, in which its production was independent of LPS effect. Conclusion: TLR4 activation by bacterial components on hAECs may be a potential risk factor for pregnancy complications

Evaluation of Antibacterial Activity and Total Phenol Compounds of Punica granatum Hydro-Alcoholic Extract

Background & Objectives: Punica granatum is a non-productive form of a plant and is used for the treatment of diseases in traditional medicine. In this study, we evaluate the antibacterial activity and the total phenol compounds of Punica granatum. Materials & Methods: Disk and well diffusion methods and MIC were used to evaluate the antibacterial activity of hydro-alcoholic extract on S. aureus and E. coli compared to standard commercial antibiotic disks. Measurement of phenol compounds were performed by Seevers and Daly colorimetric methods (Folin-ciocalteu indicator). Results: 35 and 29 mm inhibition zones in S. aureus and 22 and 17 mm inhibition zones in E. coli were shown by disk and well diffusion method, respectively. Also, 7.8 mg/ml concentration of extract showed the MIC points for two bacteria. Phenol compound of extract was 233.15&plusmn;5.1 mg/g of extraction. Conclusion: Antibacterial effect of Punica granatum compared to antibiotics indicates the strong activity against examined bacteria. Extensive antibacterial study of Punica granatum is suggested

Targeted delivery of a short antimicrobial peptide (CM11) against Helicobacter pylori gastric infection using concanavalin A-coated chitosan nanoparticles

Abstract Helicobacter pylori is the cause of most cases of stomach ulcers and also causes some digestive cancers. The emergence and spread of antibiotic-resistant strains of H. pylori is one of the most important challenges in the treatment of its infections. The present study aims to develop a concanavalin A (ConA) coated chitosan (CS) nanocarrier-based drug delivery for the targeted release of peptides to the site of H. pylori infection. Accordingly, chitosan was used as an encapsulating agent for CM11 peptide delivery by applying ionotropic gelation method. Con-A was used for coating CS nanoparticles to target H. pylori. The CS NPs and ConA-CS NPs were characterized by FTIR, dynamic light scattering (DLS), and scanning electron microscopy (SEM). The MIC of CM11-loaded ConA-CS NPs against H. pylori SS1 strain was analyzed in vitro. In order to evaluate the treatment efficiency in vivo, a gastric infection model of H. pylori SS1 strain was established in mice and histopathological studies and IL-1β cytokine assay were performed. Based on the results, the size frequency for CS NPs and ConA-CS NPs was about 200 and 350 nm, respectively. The prepared CM11-loaded ConA-CS NPs exhibited antibacterial activity against H. pylori SS1 strain with a concentration of 32 µg/ml. The highest healing process was observed in synthesized CM11-loaded ConA-CS NPs treatments and a significant decrease in IL-1β was observed. Our findings highlight the potential of chitosan nanoparticles as a drug delivery vehicle in the treatment of gastric infection model of H. pylori SS1 strain. Graphical Abstrac

E23K POLYMORPHISM IN IRANIAN PATIENTS WITH CORONARY HEART DISEASE

Abstract &nbsp;&nbsp; BACKGROUND: It has been shown that ATP-sensitive potassium (KATP) channels play an important role in physiology of myocardial adaptation to ischemia. In cardiomyocytes, the pore-forming subunits of these channels are coded by KCNJ11 gene. It was reported that the common polymorphism E23K of this gene is associated with higher susceptibility to coronary heart disease (CHD) in Chinese patients, but no other reports are available from other ethnic groups. &nbsp;&nbsp; METHODS: Iranian patients with confirmed CHD, aged over 50 years were compared with healthy controls for allelic and genotypic frequencies of this polymorphism. Patients who did not suffer from diabetes mellitus were entered into this study if they showed coronary stenosis of &gt;&nbsp;50% in at least one artery in the angiography performed after a coronary event. The subjects and controls were matched for age, gender, blood glucose, body mass index and smoking. &nbsp;&nbsp; RESULTS: No association could be found between CHD and frequencies of G and A alleles, single genotype frequencies of AA, AG, GG, and combine genotypes frequencies of AG+AA versus GG, and AG+GG versus AA. &nbsp;&nbsp; CONCLUSION: This study did not find any association between coronary heart disease and E23K polymorphism in Iranian patients. But, this finding is not conclusive due to limitation of sample size. A subsequent study with a larger sample size is recommended. &nbsp; &nbsp;&nbsp; Keywords: Single Nucleotide Polymorphism, Kir6.2 channel, KATP Channels, Caucasian. &nbsp;</p

Antibacterial effects of hydro-alcoholic extracts of Ziziphora tenuior, Teucrium polium, Barberis corcorde and Stachys inflate

Introduction: Many people are used medicinal plants due to natural, low risks and complications, andlow costs as compared with the synthetic drugs. Incidence of drug resistance against chemicalantimicrobial drugs has led the use of medicinal plants for treatment of infections in recent years. The aimof this study was to examine possible antibacterial effects of four herbal plants in vitro.Materials and Methods: Four species of domestic plants from Fars- Fasa (Iran) including Ziziphora,Stachys, Teucrium and Barberry were collected in spring and dried. Hydro-alcoholic extracts of theseplants were prepared using standard methods. Antibacterial effects were examined with disk diffusionmethod and serial broth dilution. To standardize of of study, we used antibiogram disks and ATCCbacteria.Results: Teucrium extract with 1/8 dilution and Barberry extract with 1/4 dilution showed antibacterialeffects in serial dilution method. Moreover, by using the disk diffusion method, antibacterial effects ofboth mentioned extracts against E.coli ATCC 25922 and S.aureus ATCC 25923،were found as comparedamoxicillin, ciprofloxacin, vancomycin and imipenem.Conclusion: The results of this study and other studies show that extracts of herbal plants, instead ofchemical drugs, can be used to treat infections. Of course, before using them all their side effects shouldbe carefully checked in in vitro and in vivo studies