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Zohre Sadeghi,1 Mahdi Fasihi-Ramandi,2 Saeid Bouzari1 1Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran; 2Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IranCorrespondence: Saeid BouzariDepartment of Molecular Biology, Pasteur Institute of Iran, Tehran, IranTel +98 21 6696 9297Fax +98 21 66492619Email bouzari@pasteur.ac.irIntroduction: Vaccine formulation with appropriate adjuvants is an attractive approach to develop protective immunity against pathogens. Calcium phosphate nanoparticles (CaPNs) are considered as ideal adjuvants and delivery systems because of their great potential for enhancing immune responses. In the current study, we have designed nanoparticle-based vaccine candidates to induce immune responses and protection against B. melitensis and B. abortus.Materials and Methods: For this purpose, we used three Brucella antigens (FliC, 7α-HSDH, BhuA) and two multi-epitopes (poly B and poly T) absorbed by CaPNs. The efficacy of each formulation was evaluated by measuring humoral, cellular and protective responses in immunized mice.Results: The CaPNs showed an average size of about 90 nm with spherical shape and smooth surface. The CaPNs-adsorbed proteins displayed significant increase in cellular and humoral immune responses compared to the control groups. In addition, our results showed increased ratio of specific IgG2a (associated with Th1) to specific IgG1 (associated with Th2). Also, immunized mice with different vaccine candidate formulations were protected against B. melitensis 16M and B. abortus 544, and showed same levels of protection as commercial vaccines (B. melitensis Rev.1 and B. abortus RB51) except for BhuA-CaPNs.Discussion: Our data support the hypothesis that these antigens absorbed with CaPNs could be effective vaccine candidates against B. melitensis and B. abortus.Keywords: calcium phosphate nanoparticle, adjuvant, Brucella, vaccine, immune response

Morteza Abkar,1 Mahdi Fasihi-Ramandi,2 Hamid Kooshki,3 Abbas Sahebghadam Lotfi4 1Nanomedicine and Nanobiology Research Center, Shiraz University of Medical Sciences, Shiraz, 2Molecular Biology Research Center, 3Nanobiotechnology Research Center, Baqiyatallah University of Medical Sciences, 4Department of Clinical Biochemistry, Faculty of Medicine, Tarbiat Modares University, Tehran, Iran Abstract: Brucellosis is a group of closely associated zoonotic bacterial illnesses caused by members of the genus Brucella. B. melitensis Omp31 is a promising candidate for a subunit vaccine against brucellosis. This study surveyed the immunogenicity of Omp31 alone and with incomplete Freund’s adjuvant (Omp31-IFA) and N-trimethyl chitosan (TMC/Omp31) nanoparticles (NPs), as well as the effect of Omp31 immunization route on immunological responses and protection. After expression and purification, the recombinant Omp31 (rOmp31) was loaded onto TMC NPs by ionic gelation. The particle size, loading efficiency and in vitro release of the NPs were examined. Omp31-IFA was administered intraperitoneally, while TMC/Omp31 NPs were administered orally and intraperitoneally. According to the antibody subclasses and cytokine profile, intraperitoneal immunization by Omp31-IFA and TMC/Omp31 NPs induced T helper 1 (Th1) and Th1–Th2 immune responses, respectively. On the other hand, oral immunization with TMC/Omp31 NPs elicited a mixed Th1–Th17 immune response. Data obtained from the cell proliferation assay showed that vaccination with Omp31 stimulated a vigorous antigen-specific cell proliferative response, which could be further increased after oral immunization with TMC/Omp31 NPs. Vaccinated groups of mice when challenged with B. melitensis 16M were found to be significantly protected in the orally administered group in comparison with the intraperitoneally immunized mice. Results of this study indicated that the reason for high protection after oral vaccination can be via elicited Th17 response. Keywords: brucellosis, Th17, trimethyl chitosan, vaccine, nanoparticle

Background & Objectives: Ecologic conditions affect production of effective pharmaceutical products. We evaluate the effect of ecologic conditions on antibacterial effects of herbal plants in Fars. Materials & Methods: We collected eight plant species including Trachyspermum copticum, Withania somnifera, Laurus nobilis, Carum carvi, Thymus, Rosemary, Zataria multiflora, and Lavandula in Fasa and Sepidan. Their antibacterial effects were studied by disk diffusion method, well assay method, and Minimum Inhibitory Concentration (MIC). Ecological and weather conditions were derived from internet. Results: Comparing with standard antibacterial indexes the following results were achieved. Fasa herbs including Trachyspermum copticum, Zataria multiflora, Laurus nobilis, and Rosemary had the most effect on Staphylococcus aureus, while Carum carvi had the least effect. This effect in Sepidan herbs was the most in Trachyspermum copticum, Zataria multiflora, and Rosemary, while Carum carvi had the least effect. Surveying the effects against E.coli among Fasa herbs, Zataria multiflora had the most and Carum carvi and Lavandula had the least effects. On the other hand, among Sepidan herbs, Trachyspermum had the highest and Carum carvi, Lavandula, and Laurus nobilis had the lowest effects on E.coli. Antibacterial effects of Fasa herbs against Pseudomonas aeruginosa was the most in Zataria multiflora and Thymus and the least in Carum carvi. In Sepidan herbs, it was the highest in Zataria multiflora and the lowest in Carum carvi. Conclusion: Considering the differences in antibacterial effects of the herbs in two ecologic conditions, we suggest other researchers to pay attention to the mentioned effects in their studies. [ABSTRACT FROM AUTHOR]

Background & Objective: Nanomedicine delivery systems are known as potent therapeutictools. In addition to possessing an effective agent of turmeric, Curcumin shows antibacterial properties as well. Curcumin is not water-soluble and it can be solved in water via nanotechnology-base methods. Chitosan is a natural and biodegradable substance that is utilized for the Production of Nanoparticles (NPs) carrying drug. In the following investigation, curcumin is loaded in chitosan NPs and ultimately, the resulting nano-drug is studied as an antibacterial agent. Materials & Methods: In this study, NPs are produced using chitosan and Tripolyphosphate (TPP) salt. Curcumin solution was loaded in chitosan NPs during their production. Next, the skins of BALB/c mice infected with staphylococcus aureus are treated by curcumin-loaded chitosan NPs for 3 days. Afterwards, in order to evaluate the antibacterial property of the nano-drug, these skin suspensions of mice are cultured in bacterial medium. Results: Dynamic Light Scattering (DLS) reveals the charge of + 7 ± 2 mV and the size of 160 ± 10 nm in curcuminloaded chitosan NPs. Moreover, Transmission electron microscopy (TEM) and atomic force microscopy (AFM) indicates a spiral shape. Therefore, the evaluation of the optical density by spectrophotometry demonstrates that 75 ± 2 % of curcumin are loaded in chitosan NPs. Bacterial culture shows that curcumin-loaded chitosan NPs significantly inhibited staphylococcus aureus growth. Conclusion: This study demonstrates that curcumin-loaded chitosan NPs can be applied as a potent agent in treatment of bacterial skin infections. [ABSTRACT FROM AUTHOR]

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.

Background: Toxoplasmosis is a cosmopolitan infectious disease in warm-blooded mammals that poses a serious worldwide threat due to the lack of effective medications and vaccines., Aims: The purpose of this study was to design a multi-epitope vaccine using several bioinfor-matics approaches against the antigens of Toxoplasma gondii (T. gondii)., Methods: Three proteins of T. gondii, including ROP18, MIC4, and SAG1, were analyzed to predict the most dominant B- and T-cell epitopes. Finally, we designed a chimeric immunogen RMS (ROP18, MIC4, and SAG1) using some domains of ROP18 (N377-E546), MIC4 (D302-G471), and SAG1 (T130-L299) linked by rigid linker A (EAAAK) A. Physicochemical prop-erties, secondary and tertiary structures, antigenicity, and allergenicity of RMS were predicted utilizing immunoinformatic tools and servers., Results: RMS protein had 545 amino acids with a molecular weight (MW) of 58,833.46 Da and a theoretical isoelectric point (IP) of 6.47. The secondary structure of RMS protein con-tained 21.28% alpha-helix, 24.59% extended strand, and 54.13% random coil. In addition, eval-uation of antigenicity and allergenicity showed the protein to be an immunogen and non-aller-gen. The results of the Ramachandran plot indicated that 76.4%, 12.9%, and 10.7% of amino acid residues were incorporated in the favored, allowed, and outlier regions, respectively. ΔG of the best-predicted mRNA secondary structure was -593.80 kcal/mol, which indicated that a stable loop was not formed at the 5' end., Conclusion: Finally, the accuracy and precision of the in silico analysis must be confirmed by successful heterologous expression and experimental studies., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Brucella is a facultative intracellular gram-negative coccobacillus. It is nonsporulating and reproduced in macrophage phagosomes. The use of nanostructures as drug and vaccine carriers has recently received attention due to their ability to control the release profile and protect the drug molecules. This study presents a suitable nano-polyethyleneimine formulation to be used as an immunoadjuvant and LPS along with trivalent candidate antigens of TF, BP26, and omp31 to selectively stimulate the immune response. After designing and evaluating the immunogenic structure by databases and bioinformatics software, recombinant protein cloning and gene expression were performed in Escherichia coli BL21 bacteria. This protein was extracted from the cultured cells, purified by Ni-NTA column. After placing the antigen inside the polyethyleneimine nanostructure, various properties of the nanoparticles, including their size, zeta potential, and retention rate for injection and inhalation of mice, diffusion efficacy, and antigen binding evaluation were evaluated. Mice were treated with different groups of antigens and nanoparticles on days 0, 10, 24, and 38. Two weeks after the last injection, the level of cytokines were investigated in spleen cells, including IFN-γ, IL-4, and IL-12. The serum concentration of IgG2a and IgG1 antibodies were also assessed. The response was consistent with significant production of IgG1, IgG2a, IFN-γ21, IL-12, and IL-4 compared to the controls (P < 0.05). Compared to the positive and negative control groups, recombinant protein and nanoparticles showed a good response in subsequent injections with live bacterial strains. The present study also revealed the potential of the developed recombinant protein as a candidate in the design and manufacture of subunit vaccines against Brucella species. This protein stimulates cellular and humoral immune responses compared to the positive control groups. These findings can be useful in the prevention and control of brucellosis and pave the way for further research by researchers around the world., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Purpose: Sulfur mustard (SM) is a potent blistering agent. This alkylating chemical agent has extremely toxic effects on the eye. MMP-2 and MMP-9 are the two most important matrix metalloproteinase enzymes involved in the pathology of chemical eye injuries. Curcumin is regarded as a natural anti-inflammatory agent. This study aims to compare the anti-inflammatory effects of curcumin versus doxycycline on chemically induced corneal injuries., Methods: The HCE-2 cell line was used as a model for corneal cells. The effective concentrations of 2-chloroethyl ethyl sulfide (CEES) - as an analog of SM - doxycycline, and curcumin were determined using the MTT assay. The gene expression of MMP-2 , MMP-9 , and tissue inhibitors of metalloproteinase (TIMP-1) was evaluated by the real-time PCR method. Also, the activity of MMP-2 and MMP-9 enzymes was determined by zymography., Results: The expression of the MMP-2 and MMP-9 genes increased 5- and 3.3-fold after exposure to CEES, respectively. Following the treatment with curcumin and doxycycline, MMP-2 expression decreased significantly. Also, after treatment with curcumin and doxycycline, the MMP-9 expression decreased 2.5- and 1.6-fold, respectively. The reduction in activity was 32% for MMP-2 and 56% for MMP-9 after treatment with curcumin. The corresponding values were 12% and 40% following doxycycline treatment. There was no significant difference between the effects of curcumin and doxycycline on reducing MMP-2 expression, but the difference was statistically significant in the case of MMP-9., Conclusion: Doxycycline and curcumin can inhibit MMP expression and activity in chemically exposed corneal cells. Curcumin has a greater ability than doxycycline to inhibit MMP-2 and MMP-9 enzymes; however, the difference is statistically significant only in the case of MMP-9. After further validation, these substances can be introduced as anti- inflammatory agents to treat corneal chemical burns., Competing Interests: None., (Copyright © 2024 Zareh et al.)

Purpose: Brucellosis, a zoonotic infectious disease, is a worldwide health issue affecting animals and humans. No effective human vaccine and the complications caused by the use of animal vaccines are among the factors that have prevented the eradication of the disease worldwide. However, bio-engineering technologies have paved the way for designing new targeted and highly efficacious vaccines. In this regard, the study aimed to evaluate immunity induced by mannosylated niosome containing Brucella recombinant trigger factor/Bp26/Omp31 (rTBO) chimeric protein in a mouse model., Materials and Methods: rTBO as chimeric antigen (Ag) was expressed in Escherichia coli BL21 (DE3) and, after purification, loaded on niosome and mannosylated niosome. The characteristics of the nanoparticles were assessed. The mice were immunized using rTBO, niosome, and mannosylated niosome-rTBO in intranasal and intraperitoneal routes. Serum antibodies (immunoglobulin [Ig]A, IgG, IgG1, and IgG2a) and splenocyte cytokines (interferon-gamma, interleukin [IL]-4, and IL-12) were evaluated in immunized mice. Finally, immunized mice were challenged by B. melitensis and B. abortus . A high antibody level was produced by niosomal antigen (Nio-Ag) and mannosylated noisomal antigen (Nio-Man-Ag) compared to the control after 10, 24, and 38 days of immunization. The IgG2a/IgG1 titer ratio for Nio-Man-Ag was 1.2 and 1.1 in intraperitoneal and intranasal methods and lower than one in free Ag and Nio-Ag. Cytokine production was significantly higher in the immunized animal with Ag-loaded nanoparticles than in the negative control group (p<0.05). Moreover, cytokine and antibody levels were significantly higher in the injection than in the inhalation method (p<0.05)., Results: The combination of mannosylated noisome and rTBO chimeric proteins stimulate the cellular and humoral immune response and produce cytokines, playing a role in developing the protective acquired immune response in the Brucella infectious model. Also, the intraperitoneal route resulted in a successful enhancement of cytokines production more than intranasal administration., Conclusion: Designing an effective vaccine candidate against Brucella that selectively induces cellular and humoral immune response can be done by selecting a suitable nanoniosome formulation as an immunoadjuvant and recombinant protein as an immune response-stimulating Ag., Competing Interests: No potential conflict of interest relevant to this article was reported., (© Korean Vaccine Society.)

Exposure to aflatoxin B1 can be associated with reproductive toxicity, accompanied by decreased sperm concentration in animal models. The aim of this meta-analysis was to determine the correlation between aflatoxin B1 exposure and sperm concentrations of male rodents (both mice and rats). According to inclusion and exclusion criteria, 8 articles were selected to assess in the current meta-analysis. The random effects and pooled analysis indicated that sperm concentration was decreased in mice [MD sperm = -20.79×10 6 /sperm/g testis (95%CI =-1.3 to -50.5)] and in rats [-24.34×10 6 /sperm/g testis (95%CI: -7.60 to -44.35)] after exposure to aflatoxin B1 compared with control groups. A significant heterogeneity was found among studies (for mice I 2 =99.7%, %, P <0.000 and rats =I 2 =98.8, P <0.000). The findings of present meta-analysis showed the association between aflatoxin B1 exposure and a decrease in sperm concentration in rodents.

Purpose: Due to the many problems with commercially available vaccines, the production of effective vaccines against brucellosis is a necessity. The aim of this study was to evaluate the immune responses caused by the chimeric protein consisting of trigger factor, Bp26, and Omp31 (TBO) along with aluminum hydroxide (AH/TBO) and selenium (Se/TBO) nanoparticles (NPs) as adjuvants in mouse model., Materials and Methods: Recombinant antigen expression was induced in Escherichia coli BL21 (DE3) bacteria using IPTG (isopropyl-d-1-thiogalactopyranoside). Purification and characterization of recombinant protein was conducted through NiFe 3 O 4 NPs, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and Western blot. NP characteristics, including morphology and particle size, were measured in vitro . The recombinant TBO was loaded on to AH and Se NPs and were administered subcutaneously. After mice immunization, measurement of antibody titter and protection assay was performed., Results: The average sizes of AH and Se NPs were about 60 nm and 150 nm, respectively. The enzyme-linked immunosorbent assay results showed that the serum of mice immunized by subcutaneous injection with both nanovaccines produced significant immunoglobulin G (IgG) responses against the chimeric antigen. The results of TBO-specific IgG isotype (IgG2a/IgG1) analysis showed that both AH and Se NPs induced a type to T-helper immune response. In addition, the results of the challenge with the pathogenic strain of Brucella melitensis 16M showed that vaccinated mice with AH/TBO NPs indicated a higher reduction of bacterial culture than immunized mice with Se/TBO NPs and TBO alone., Conclusion: The results showed that AH NPs carrying chimeric antigen can be a promising vaccine candidate against brucellosis by producing protective immunity., Competing Interests: No potential conflict of interest relevant to this article was reported., (© Korean Vaccine Society.)

Toxoplasmosis is a zoonotic disease that infects most animals, including humans. Pyrimethamine/sulfadiazine is the standard treatment for toxoplasmosis. Although this treatment has been successful, it is often associated with side effects that cannot be tolerated. Therefore, various compounds have been proposed as alternative treatments for toxoplasmosis. Antimicrobial peptides (AMPs) act on various pathogens, from viruses to protozoa. The purpose of the present study was to evaluate the effects of CM11 on in vitro and in vivo Toxoplasma gondii infection. For in vitro experiments, VERO cells were treated with different concentrations of CM11 (1-128 μg/ml) compared to sulfadiazine (SDZ) (0.78-100 μg/ml). MTT and lactate dehydrogenase (LDH) assays evaluated the cell viability and plasma membrane integrity. Then, the inhibitory concentration (IC 50 ) values were determined for treating tachyzoites of T. gondii before or on cells previously infected. Annexin V-FITC/propidium iodide (PI) staining was used to distinguish viable and apoptotic cells. The effect of CM11, SDZ, and a combination of CM11 and SDZ was evaluated in the BALB/c mouse model of acute toxoplasmosis. CM11 was effective on tachyzoites of T. gondii and had a time and dose-dependent manner. The results of the MTT assay showed that the CC 50 values of CM11 and SDZ were estimated at 17.4 µg/ml and 62.3 µg/ml after 24-h, respectively. The inhibitory concentration (IC 50 ) of CM11 and SDZ on infected cells was estimated at 1.9 µg/ml and 1.4 µg/ml after 24-h, respectively. The highest rate of apoptosis (early and late) in high concentrations of SDZ and CM11 was determined for tachyzoites (2.13 % and 13.88 %), non-infected VERO cells (6.1 % and 19.76 %), and infected VERO cells (7.45 % and 29.9 %), respectively. Treating infected mice with CM11 and a combination of CM11 and SDZ had increased survival time. Based on the mentioned results, it can be concluded that CM11 has a beneficial effect on tachyzoites of T. gondii in vitro. The result of the mouse model suggests that CM11, either alone or in combination with other chemotherapeutic agents, could be a potential therapeutic for toxoplasmosis. Hence, antimicrobial peptides could be applied as promising anti-toxoplasma agents for treating toxoplasmosis., Competing Interests: Declaration of Competing Interest The authors declared no potential conflicts of interest concerning the research or authorship., (Copyright © 2023 Elsevier B.V. All rights reserved.)

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., (© 2023. The Author(s).)

Background: We aimed to design a B and T cell recombinant protein vaccine of Toxoplasma gondii with in silico approach. MIC13 plays an important role in spreading the parasite in the host body. GRA1 causes the persistence of the parasite in the parasitophorous vacuole. SAG1 plays a role in host-cell adhesion and cell invasion., Methods: Amino acid positions 73-272 from MIC13, 71-190 from GRA1, and 101-300 from SAG1 were selected and joined with linker A(EAAAK)A. The structures, antigenicity, allergenicity, physicochemical properties, as well as codon optimization and mRNA structure of this recombinant protein called MGS1, were predicted using bioinformatics servers. The designed structure was synthesized and then cloned in pET28a (+) plasmid and transformed into Escherichia coli BL21., Results: The number of amino acids in this antigen was 555, and its antigenicity was estimated to be 0.6340. SDS-PAGE and Western blotting confirmed gene expression and successful production of the protein with a molecular weight of 59.56kDa. This protein will be used in our future studies as an anti- Toxoplasma vaccine candidate in animal models., Conclusion: In silico methods are efficient for understanding information about proteins, selecting immunogenic epitopes, and finally producing recombinant proteins, as well as reducing the time and cost of vaccine design., Competing Interests: Conflict of Interest The authors declare that there is no conflict of interest., (© 2023 Hosseininejad et al. Published by Tehran University of Medical Sciences.)

Despite the huge efforts of microbiologists, infectious diseases have yet remained one of the leading causes of death in humans, further highlighting the research priority for controlling opportunistic pathogens. Many researchers have used antibacterial peptides to solve the problem of antibiotic resistance. This research is thus conducted to investigate the antibacterial and anti-biofilm activity of a novel modified cecropin-melittin 11-peptide with improved therapeutic properties and lower side effects. After synthesis and purification of mCM11 (NH2-WRLFRRILRVL-NH2) by solid-phase synthesis and HPLC methods, respectively, the antibacterial and biofilm inhibitory activities were explored in vitro. TMHMM was used to confirm the reaction of mCM11 on the plasma membrane of the prokaryotic cells. The interaction between mCM11 on Acinetobacter baumannii strains was investigated by molecular docking using ClusPro2.0. Hemolysis and therapeutic indexes were also calculated to quantify the relative safety and adverse effects of mCM11. According to the results, mCM11 has a high inhibitory and lethal effect on A. baumannii strains due to its cationic properties and new specific sequence. Molecular docking revealed the release of a significant amount of energy when mCM11 binds to the surface of A. baumannii in an appropriate site. The findings indicated that mCM11 IC50 (4 μg/mL) lysed 2.78% of RBCs; moreover, 8 strains of Acinetobacter baumannii showed a favorable therapeutic index. The mCM11 exhibits strong antibacterial and antibiofilm activities against A. baumannii strains, suggesting its potential therapeutic role in infections caused by these strains. Similar to its impact on A. baumannii, mCM11 could be a suitable alternative to antibiotics in combat against antibiotic-resistant bacteria in the in vivo experiments., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

One promising approach to increase protection against infectious diseases is to use adjuvants that can selectively stimulate the immune responses. In this study, multi-epitope antigens associated with LPS loaded chitosan (LLC) as toll-like receptor agonist or mannosylated chitosan nanoparticle (MCN) as vaccine delivery system were evaluated for their ability to stimulate immune responses to Brucella infection in mice model. Our results indicated that the addition of MCN to our vaccine formulations significantly elicited IFN-γ and IL-2 cytokines and antibody titers, in comparison with the non-adjuvanted vaccine candidates. The present results indicated that multi-epitopes and their administration with LLC or MCN induced Th1 immune response. In addition, vaccine candidates containing MCN provided high percentage of protection against B. melitensis and B. abortus infection. Our results provided support to previous reports indicating that MCNs are attractive adjuvants and addition of this adjuvant to multi-epitopes antigens play an important role in the development of vaccine against Brucella., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Inc.)

Many recent studies have been conducted to find new DNA vaccines based on Toxoplasma gondii antigens. DNA vaccines encoding complex of different antigens showed better immune responses compared to single antigen vaccine. In this study, we constructed a DNA vaccine encoding SAG1, SAG3, MIC4, GRA5, GRA7, AMA1 and BAG1 against T. gondii, and evaluated the immune response it induced in BALB/c mice. For this purposes, thirty BALB/c mice were randomly divided into three groups containing tenmice each. There were two negative control groups (PBSand pVAX1 vector) and one vaccination group (pVAX1-MAF, Multantigenic Fragment). On days 0, 14 and 28, the mice were immunized intramuscularly, and 5 weeks later they were challenged with T. gondii RH strain. The immune responses were evaluated using lymphocyte proliferation assay, T-cell subsets detection, and measurement of antibody and cytokine levels. The results showed that mice immunized with pVAX1-MAF developed high levels of IL-2, IL-12, IgG and IFN- γ as well as CD3 + CD4 + T cells. In addition, the survival time of mice immunized by pVAX1-MAF was longer than that control mice. In conclusion, our results show that the multiple DNA vaccine encodingSAG1, SAG3, mic4, GRA5, GRA7, AMA and BAG1effectively enhanced humoral and cellular immune responses, and prolonged the survival time. Together this would suggest that further investigation may result in a promising candidate vaccine to treat toxoplasmosis., (Copyright © 2022. Published by Elsevier Inc.)

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 × 10 3 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., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2022. Published by Elsevier Inc.)

Regulfatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs) are common immunosuppressive cells in the tumor microenvironment. These cells, through various mechanisms, inhibit antitumor immune responses and impede effective therapies. Therefore, designing an efficient protocol for inducing immune surveillance in tumors is highly recommended. Recently, nanoliposomes have provided broad-spectrum and state-of-the-art vehicles to deliver antigens or immune system compartments in immunotherapies. It has been shown that different lipids in the structure of liposomes and various liposomal formulations can affect immune responses in the tumor microenvironment. This study was aimed to evaluate the effects of four different liposomal formulations on MDSCs and Tregs in C26 tumor-bearing mice. To this end, after preparing liposomes, they were injected into tumor-inoculated mice and analyzed MDSC and Treg population and functions in spleen and tumor tissues. Results showed that 1,2-dioleoyl-3-trimethylammonium propane (DOTAP)-containing liposomes reduced MDSC population and activity in the spleen, but not tumor, compared with other groups significantly (p < 0.05 and p < 0.01, respectively). Moreover, DOTAP-containing liposomes reduced the expression of S100A8 and arginase-1 genes in splenic MDSCs (p < 0.05). In conclusion, we provided evidence that DOTAP-containing liposomes contributed to stimulating immune responses and provided a situation to inhibit immunosuppression in the tumor microenvironment., (© 2021 International Union of Biochemistry and Molecular Biology, Inc.)

Objectives: Dental implants are a known treatment today. It is necessary to have at least 2 mm of bone around the implant, especially in the buccal aspect of the anterior maxilla (esthetic zone). Some systemic conditions, such as menopause, can affect the body's bone mass as well as the alveolar bone. Considering that few studies have been carried out on the effect of menopause on the thickness and topography of alveolar bone, we decided to investigate the effect of menopause on buccal alveolar bone thickness in the anterior maxillary teeth in menopausal women. MATERIAL AND METHODS: In this descriptive-analytical cross-sectional study, two subgroups of menopausal women and nonmenopausal women were considered. Data were extracted from 30 patients referred to a private radiology center in Mashhad for CBCT imaging. In addition, the buccal bone thickness in the crest and middle areas of the anterior maxillary teeth was measured and the difference between the two groups was investigated. The buccal bone thickness of the aesthetic area was evaluated with CBCT Planmeca ProMax 3D Max (Planmeca) by Planmeca Romexis 5.3.4 software, with 200 μm Voxel size and Fov 90 × 60 mm., Results: In this study, 30 women with a mean age of 49.75 ± 3.65 years in the nonmenopausal and menopausal groups were examined. It was found that the mean buccal bone thickness of the anterior maxilla in the nonmenopausal group (0.65 ± 0.25 mm) was higher than in the menopausal group (0.56 ± 0.20 mm), but the difference was not statistically significant (p = .2999). Only in the crestal bone of the right canine, the average bone thickness in nonmenopausal group (0.77 ± 0.33 mm) was significantly higher than the menopausal group (0.49 ± 0.22 mm) (p = .011)., Conclusions: Owing to changes in the volume and thickness of alveolar bone in menopausal women, the thickness of the buccal bone in the aesthetic area decreases, but this is not statistically significant., (© 2022 The Authors. Clinical and Experimental Dental Research published by John Wiley & Sons Ltd.)

Efferocytosis is the process by which apoptotic cells are removed without inflammation to maintain tissue homeostasis, prevent unwanted inflammatory responses, and inhibit autoimmune responses. Coordination of efferocytosis occurs via many surfaces and chemotactic molecules and adaptors. Recently, soluble positive or negative mediators of efferocytosis, have been more noticeable as non-invasive valuable biomarkers in prognosis and targeted therapy. These soluble factors can be detected in different bodily fluids, such as serum, plasma, and urine as a non-invasive method. There are lots of studies that have tried to show the importance of receptors and ligands in disorders; while a few studies tried to indicate the importance of soluble forms of receptors/ligands and their clinical aspects as a systemic compound and shedding of targets related to efferocytosis. Some of these soluble forms also can be as sensitive as specific biomarkers for certain diseases compared with routine biomarkers, such as soluble circulatory Lectin-like oxidized low-density lipoprotein receptor-1 vs. troponin T in the acute coronary syndrome. Thus, this review tried to gain more understanding about efferocytosis-related unwanted soluble receptors/ligands, their roles, the clinical significance, and potential for diagnosis, and prognosis related to different diseases., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

It has been about a century since the discovery of the first antibiotic, and during this period, several antibiotics were produced and marketed. The production of high-potency antibiotics against infections led to victories, but these victories were temporary. Overuse and misuse of antibiotics have continued to the point that humanity today is almost helpless in the fight against infection. Researchers have predicted that by the middle of the new century, there will be a dark period after the production of antibiotics that doctors will encounter antibiotic-resistant infections for which there is no cure. Accordingly, researchers are looking for new materials with antimicrobial properties that will strengthen their ammunition to fight antibiotic-resistant infections. One of the most important alternatives to antibiotics introduced in the last three decades is antimicrobial peptides (AMPs), which affect a wide range of microbes. Due to their different antimicrobial properties from antibiotics, AMPs can fight and kill MDR, XDR, and colistin-resistant bacteria through a variety of mechanisms. Therefore, in this study, we intend to use the latest studies to give a complete description of AMPs, the importance of colistin-resistant bacteria, and their resistance mechanisms, and represent impact of AMPs on colistin-resistant bacteria. KEY POINTS: • AMPs as limited options to kill colistin-resistant bacteria. • Challenge of antibiotics resistance, colistin resistance, and mechanisms. • What is AMPs in the war with colistin-resistant bacteria?, (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

A promising strategy for controlling animal brucellosis is vaccination with commercial vaccine strains (Brucella melitensis Rev.1 and Brucella abortus RB51). Owing to safety concerns associated with these vaccines, developing a more effective and safe vaccine is essential. In this study, we examined the capacity of BhuA, 7α-HSDH or FliC antigens in the presence or absence of adjuvant in eliciting immune responses against brucellosis. After cloning, expression and purification, these proteins were used to examine immunologic responses. All immunized mice induced a vigorous IgG, with a predominant IgG2a response. Moreover, splenocytes of immunized mice proliferated and produced IL-2 and IFN-γ, suggesting the induction of cellular immunity. The high IgG2a/IgG1 ratio and IL-2 and IFN-γ indicated a Th1-oriented immune response in test groups. BhuA-, 7α-HSDH- or FliC- poly I:C formulations were the most effective at inducing Th1 immune response compared to groups immunized with naked proteins. Immunization with proteins protected mice against B. melitensis 16M and B. abortus 544. The proteins in adjuvant induced higher levels of protection than proteins only and exhibited similar degree of protection to live attenuated vaccines. Our results, for first time, introduced five potential candidates for subunit vaccine development against B. melitensis and B. abortus infection., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Melatonin has been known as an anti-inflammatory agent and immune modulator that may address progressive pathophysiology of coronavirus disease 2019 (COVID-19)., Aim of the Study: To evaluate the clinical efficacy of adjuvant, use of melatonin in patients with COVID-19., Methods: This single-center, double-blind, randomized clinical trial included 74 hospitalized patients with confirmed mild to moderate COVID-19 at Baqiyatallah Hospital in Tehran, Iran, from April 25, 2020-June 5, 2020. Patients were randomly assigned in a 1:1 ratio to receive standard of care and standard of care plus melatonin at a dose of 3 mg three times daily for 14 d. Clinical characteristics, laboratory, and radiological findings were assessed and compared between two study groups at baseline and post-intervention. Safety and clinical outcomes were followed up for four weeks., Results: A total of 24 patients in the intervention group and 20 patients in the control group completed the treatment. Compared with the control group, the clinical symptoms such as cough, dyspnea, and fatigue, as well as the level of CRP and the pulmonary involvement in the intervention group had significantly improved (p <0.05). The mean time of hospital discharge of patients and return to baseline health was significantly shorter in the intervention group compared to the control group (p <0.05). No deaths and adverse events were observed in both groups., Conclusions: Adjuvant use of melatonin has a potential to improve clinical symptoms of COVID-19 patients and contribute to a faster return of patients to baseline health., Competing Interests: Competing Interests All authors have no conflict of interest to declare., (Copyright © 2021. Published by Elsevier Inc.)

In the last century, the emergence of in silico tools has improved the quality of healthcare studies by providing high quality predictions. In the case of COVID-19, these tools have been advantageous for bioinformatics analysis of SARS-CoV-2 structures, studying potential drugs and introducing drug targets, investigating the efficacy of potential natural product components at suppressing COVID-19 infection, designing peptide-mimetic and optimizing their structure to provide a better clinical outcome, and repurposing of the previously known therapeutics. These methods have also helped medical biotechnologists to design various vaccines; such as multi-epitope vaccines using reverse vaccinology and immunoinformatics methods, among which some of them have showed promising results through in vitro , in vivo and clinical trial studies. Moreover, emergence of artificial intelligence and machine learning algorithms have helped to classify the previously known data and use them to provide precise predictions and make plan for future of the pandemic condition. At this contemporary review, by collecting related information from the collected literature on valuable data sources; such as PubMed, Scopus, and Web of Science, we tried to provide a brief outlook regarding the importance of in silico tools in managing different aspects of COVID-19 pandemic infection and how these methods have been helpful to biomedical researchers., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Authors.)

SARS-CoV-2 is a corona virus that has been the cause for one of the deadliest pandemics of history, started since 2019. Suppressing the activity of the critical enzymes in the SARS-CoV-2 could potentially inhibit a vital step in viral life cycle. Papain-like protease (PLpro) could be regarded as a critical enzyme in viral replication of SARS-CoV-2. In this research, it was aimed to suppress the activity of PLpro enzyme by using potential plant-derived protease inhibitor peptides. For this purpose, 11 plant derived peptides that could potentially inhibit protease activity were selected from literature. The structures of the PLpro and the peptide ligands were acquired from PDB (protein data bank) and after structural optimization, were docked by using HADDOCK 2.4 program. Analyzing the results indicated that VcTI from Veronica hederifolia provides effective molecular interactions at both liable Zn site and classic active site of PLpro, making it a potential inhibitory ligand for this enzyme that could be used for halting the replication of SARS-CoV-2. Molecular dynamic assay confirmed that the selected receptor and ligand complex was stable. Future in vitro and in vivo investigations are required to verify the efficiency of this compound as a potential therapeutic against SARS-CoV-2 infection., Supplementary Information: The online version contains supplementary material available at 10.1007/s10989-021-10331-8., Competing Interests: Conflict of interestNone to declare., (© The Author(s), under exclusive licence to Springer Nature B.V. 2021.)

Objectives: The present in vitro study aimed to evaluate whether Lactobacillus delbrueckii and Lactobacillus rhamnosus treatments can induce regulatory phenotype, together with modulating the expression of chemokine receptors (CRs) in dendritic cells (DCs). The CRs of DCs have been found to be involved in the pathogenesis of Systemic lupus erythematosus (SLE) through directing recruitment and migration of immune cells., Materials and Methods: In brief, monocytes of patients with SLE and healthy donors were isolated and differentiated to regulatory or inflammatory mature DCs through treatment with L. delbrueckii , L. rhamnosus , mixed probiotics, and LPS., Results: FACScan analysis showed that the expression of CRs including CXCR3, CCR5, CCR4, and CCR3, was significantly reduced in all probiotic-treated groups of SLE and healthy (control) donors when compared with the LPS treated group., Conclusion: The results demonstrated that tolerogenic probiotics could prevent or decrease the expression of inflammatory CRs on the surface of tolerogenic DCs during the maturation process., Competing Interests: The authors declare that they have no conflicts of interest.

The inflammasome as a multiprotein complex has a role in activating ASC and caspase-1 resulting in activating IL-1β in various infections and diseases like corona virus infection in various tissues. It was shown that these tissues are affected by COVID-19 patients. According to the current evidence, melatonin is not veridical while possessing a high safety profile, however, it possesses indirect anti-viral actions owing to its anti-oxidation, anti-inflammation, and immune improving properties. This study aims to assess the impacts of melatonin as the complementary treatments on oxidative stress agents and inflammasome activation in patients with COVID-19. Melatonin supplement (9 mg daily, orally) was provided for the patients hospitalized with a COVID-19 analysis for 14 days. For measuring IL-10, IL-1β, and TNF-α cytokines and malondialdehyde (MDA), nitric oxide (NO), and superoxide dismutase (SOD) level and the expression of CASP1 and ASC genes, blood samples were gathered from the individuals at the start and termination of the therapy. Our findings indicated that melatonin is used as a complementary treatment to reduce the levels of TNF-α and IL-1β cytokines, MDA, and NO levels in COVID-19 patients and significantly increase SOD level, however, the levels of IL-10 cytokine possesses no considerable changes. The findings revealed that genes of CASP1 and ASC were dysregulated by melatonin regulating the inflammasome complex. Based on the findings of the current study, it is found that melatonin can be effective as a medicinal supplement in decreasing the inflammasome multiprotein complex and oxidative stress along with beneficial impacts on lung cytokine storm of COVID-19 patients., (© 2021 Walter de Gruyter GmbH, Berlin/Boston.)

Coronavirus (SARS-CoV-2) is spreading rapidly in the world and is still taking a heavy toll. Studies show that cytokine storms and imbalances in T-helper (Th)1/Th2 play a significant role in most acute cases of the disease. A number of medications have been suggested to treat or control the disease but have been discontinued due to their side effects. Melatonin, as an intrinsic molecule, possesses pharmacological anti-inflammatory and antioxidant properties that decreases in concentration with age; as a result, older people are more prone to various diseases. In this study, patients who were hospitalized with a diagnosis of coronavirus disease 2019 (COVID-19) were given a melatonin adjuvant (9 mg daily, orally) for fourteen days. In order to measure markers of Th1 and Th2 inflammatory cytokines (such as interleukin (IL)-2, IL-4, and interferon (IFN)-γ) as well as the expression of Th1 and Th2 regulatory genes (signal transducer and activator of transcription (STAT)4, STAT6, GATA binding protein 3 (GATA3), and T-box expressed in T cell (T-bet)), blood samples were taken from patients at the beginning and end of the treatment. Adjuvant therapy with melatonin controlled and reduced inflammatory cytokines in patients with COVID-19. Melatonin also controlled and modulated the dysregulated genes that regulate the humoral and cellular immune systems mediated by Th1 and Th2. In this study, it was shown for the first time that melatonin can be used as a medicinal adjuvant with anti-inflammatory mechanism to reduce and control inflammatory cytokines by regulating the expression of Th1 and Th2 regulatory genes in patients with COVID-19., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The Coronaviridae family comprises large enveloped single-stranded RNA viruses. The known human-infecting coronaviruses; severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV), novel SARS-CoV-2, human coronavirus (HCoV)-NL63, HCoV-229E, HCoV-OC43 and HKU1 cause mild to severe respiratory infections. The viral diseases induced by mammalian and avian viruses from Coronaviridae family pose significant economic and public health burdens. Due to increasing reports of viral resistance, co-infections and the emergence of viral epidemics such as COVID-19, available antiviral drugs show low or no efficacy, and the production of new treatments or vaccines are also challenging. Therefore, demand for the development of novel antivirals has considerably increased. In recent years, antiviral peptides have generated increasing interest as they are from natural and computational sources, are highly specific and effective, and possess the broad-spectrum activity with minimum side effects. Here, we have made an effort to compile and review the antiviral peptides with activity against Coronaviridae family viruses. They were divided into different categories according to their action mechanisms, including binding/attachment inhibitors, fusion and entry inhibitors, viral enzyme inhibitors, replication inhibitors and the peptides with direct and indirect effects on the viruses. Reported studies suggest optimism with regard to the design and production of therapeutically promising antiviral drugs. This review aims to summarize data relating to antiviral peptides particularly with respect to their applicability for development as novel treatments., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Glucose-regulated protein 78 (GRP78) is an endoplasmic reticulum (ER) chaperone that has been shown that is overexpressed in cancer cells. Overexpression of GRP78 on cancer cells makes this molecule a suitable candidate for cancer detection and targeted therapy. VHH is the binding fragment of camelid heavy-chain antibodies also known as "nanobody." The aim of this study is to isolate and produce a new recombinant nanobody using phage display technique to detect cancer cells. Using the c-terminal domain of GRP78 (CGRP) as an antigen, four rounds of biopanning were performed, and high-affinity binders were selected by ELISA. Their affinity and functionality were characterized by surface plasmon resonance (SPR) cell ELISA and immunocytochemistry. A unique nanobody named V80 was purified. ELISA and SPR showed that this antibody had high specificity and affinity to the GRP78. Immunofluorescence analysis showed that V80 could specifically bind to the HepG2 and A549 cancer cell lines. This novel recombinant nanobody could bind to the cell surface of different cancer cells. After further evaluation, this nanobody can be used as a new tool for cancer detection and tumor therapy., (© 2020 International Union of Biochemistry and Molecular Biology, Inc.)

Toxoplasma gondii is an intracellular apicomplexan parasite, which can cause a serious infectious disease in pregnant women and immunocompromised individuals. Therefore, the development of a polyvalent vaccine consisting of all stages of the parasite life cycle using the epitopes from tachyzoites, bradyzoites, and sporozoites is likely to be required for complete protective immunity. In this study, we designed protein vaccine candidate based on the prediction of specific epitopes (i.e., B cell and T cell) from three Toxoplasma gondii antigens. The MRS protein (MIC3: 30-180, ROP8: 85-185, and SAG1: 85-235) was expressed in Escherichia coli, and purification was performed using a HisTrap HP column and then we evaluated immunogenicity and protective property in BALB/c mice. Seventy-two mice were randomly divided into six groups, including three vaccinations (i.e., MRS, MRS-Freund, and MRS-Calcium Phosphate Nanoparticles (MRS-CaPNs)) and three control (i.e., Phosphate-buffered saline, Freund, and CaPNs) groups. All groups were immunized three times via subcutaneous injection within three-week intervals. In the vaccination groups, the BALB/c mice were injected with 20 μg of MRS protein for the first time and 10 μg of MRS for the next two times. Antibodies, cytokines, and splenocytes proliferation in the immunized mice were assayed using the enzyme-linked immunosorbent assay. Protective efficacy was analyzed by challenging the immunized mice with T. gondii of RH strain. Antibody, cytokine, and lymphocyte proliferation assays showed that the mice immunized with MRS induced stronger humoral and T helper type 1 cell-mediated immune responses, compared to the control mice. However, co-immunization with adjuvants (i.e., Freund and CaNPs) resulted in impaired immune responses. Effective protection against the parasite achieved an increase in survival time in the immunized mice, especially in the MRS-CaNPs group. The obtained results of the present study demonstrated that multi-epitope protein vaccination, MRS, is a potential strategy against toxoplasmosis infection. In addition, the vaccine co-delivered with CaPNs could provide an important key for vaccine candidate to control T. gondii infection., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

To promote the low photocatalytic efficiency caused by the recombination of electron/hole pairs and widen the photo-response wavelength window, ZnFe 2 O 4 @ZnO-N/B/RGO and ZnFe 2 O 4 @ZnO-C 3 N 4 ternary heterojunction nanophotocatalysts were designed and successfully prepared through a sol-gel technique. In comparison to bare ZnFe 2 O 4 and ZnO, the ZnFe 2 O 4 -ZnO@N/B/RGO and ZnFe 2 O 4 @ZnO-C 3 N 4 ternary products showed highly improved photocatalytic properties in the degradation of methyl orange (MO) under ultra-violet (UV) and visible light irradiation. Various physicochemical properties of the photocatalysts were evaluated through field emission scanning electron microscopy (FESEM), energy-dispersive X-ray (EDX) analysis, X-ray diffraction (XRD), UV-visible diffuse reflectance spectroscopy (DRS), Fourier transform infrared spectroscopy (FT-IR), and vibrating sample magnetometer (VSM) techniques. The observations indicated that the ternary heterojuncted ZnFe 2 O 4 @ZnO-N/B/RGO absorbs lower energy visible light wavelengths, which is an enhancement in the photocatalytic properties of ZnFe 2 O 4 @ZnO loaded on reduced graphene oxide (RGO) nanosheets and graphite-like carbon nitride (g-C 3 N 4 ). This gives the catalyst photo-Fenton degradation properties.

Due to the broad-spectrum of antibiotic resistance, herein we investigated the possibility of using imipenemconjugated silver nanoparticles (IMP-AgNPs) against multidrug-resistant isolates of Pseudomonas aeruginosa . For this purpose, 200 clinical isolates were tested against different antibiotics to determine the antimicrobial susceptibility. To identify blaVIM and blaIMP resistance genes, PCR was used. The synthesized AgNPs and conjugants were characterized using UV-vis spectroscopy, XRD, SEM, TEM, DLS, and FTIR. The stability, drug release kinetics, cytotoxicity, hemolytic and apoptotic effects of NPs were also investigated. MIC of the imipenem, AgNPs, and conjugants were evaluated versus P. aeruginosa isolates. Finally, the effects of the IMP-AgNPs to heal burn wounds in rats was evaluated. According to the results, about 68% of isolates showed resistance to imipenem (MIC ≥ 64 μg/ml to ≥ 512 μg/ml). Analytical results verified the synthesis of AgNPs and IMP-AgNPs. A Dose-dependent decrease happened in terms of the MIC values of IMP-AgNPs were also affected by the existence of resistant genes. Low cytotoxic was observed regarding AgNPs which lead to apoptosis. The histopathological results showed a considerable epithelization in treated groups with IMPAgNPs. Accordingly, IMP-AgNPs can be considered as a powerful antibacterial agent to treat the infections caused by multidrug-resistant P. aeruginosa .

Purpose: Brucellosis as a worldwide zoonotic illness affect domestic animals and humans doesn't have any vaccine for the prevention of infection in humans yet. The aim of this study was to evaluate the specific immune response following the administration of glycine nanoparticles as adjuvant and delivery system of a chimeric antigen contained trigger factor, Omp31, and Bp26 in murine model., Materials and Methods: The chimeric antigen of Brucella was cloned and expressed in Escherichia coli ( E. coli ) BL21 (DE3). Purification and characterization of recombinant protein was conducted through Ni-NTA (nickel-nitrilotriacetic acid) agarose, SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis), and Western blot. Nanoparticle characteristics including morphology, particle size distribution, zeta potential, protein retention rate, and release rate were measured in vitro . Subsequently, nanoparticle contained antigen was administered to mice and blood sample was taken to measured the antibody level., Results: The protein retention in the nanoparticles was successfully done and the nanoparticle characteristics were appropriate. The average size of glycine particles containing antigen was about 174 nm, and the absorption of protein was approximately 61.27% of the initial value, with a release rate of approximately 70% after 8 hours. Enzyme-linked immunosorbent assay result proved that the immunized sera of mice which were administered with nano-formula contains high levels of antibodies (immunoglobulin G) against recombinant chimeric antigen and also a high level of mucosal antibody (immunoglobulin A) in the oral group, which showed a desirable immunity against Brucella ., Conclusion: The results showed that chimeric antigen-loaded glycine nanoparticles can act as a vaccine candidate for inducing the cellular and humoral immune response against brucellosis., Competing Interests: No potential conflict of interest relevant to this article was reported., (© Korean Vaccine Society.)

Background: Nowadays, the use of green tea supplements has increased. Studies have shown that green tea can have positive effects on anti-inflammatory and antioxidative factors, as well as improve aerobic performance capacity. Therefore, in this study, we investigate the effects of this supplement on inflammatory factors, total antioxidant capacity responses, and maximum oxygen uptake (VO 2 Max) of healthy young men in summer., Methods: This study is a double-blind randomized controlled trial (RCT) in which 15 young men (age 25.06 ± 2.1) were randomly assigned into the green tea (GT) and placebo groups. Subjects performed maximum aerobic exercises (shuttle run 20 m) in separate workouts (14 days) in summer. They consumed 640 mg green tea extracts or maltodextrin 90 min before exercise in a double-blind design. Blood samples were collected before and after the exercise and then evaluated in the biochemistry laboratory. In this study, one way analysis of variance (ANOVA) tests were used for the statistical analysis., Results: The results of this study show that green tea supplement significantly slowed down the increasing tumor necrosis factor alpha (TNF-α) (GT: 15.03 ± 4.31 [pg/ml], placebo: 31.38 ± 7.18 [pg/ml], [ P = 0.000]); increased the total antioxidant capacity (GT: 1.04 ± 0.06 [mm], placebo group: 0.72 ± 0.04 [mm], [ P = 0.001 VO 2 ]); and Max (GT: 44.43 ± 3.06 [ml/kg/min], placebo group: 34.88 ± 1.30 [ml/kg/min], [ P = 0.001]) in the supplement group than placebo. In addition, no significant differences in interleukin 1 beta (IL-1β) was observed between thee groups (GT: 26.86 ± 5.05 [pg/ml], placebo group: 23.47 ± 3.16 [pg/ml], [ P = 0.251])., Conclusions: The consumption of green tea supplements 90 min before aerobic exercise may decrease inflammation and oxidative stress factors and improve VO 2 Max in summer., Competing Interests: There are no conflicts of interest., (Copyright: © 2020 International Journal of Preventive Medicine.)

Several factors impact the immune responses such as the chemical nature of antigens, the physiologic and metabolic condition of the responsive cells, the site of antigen recognition, and neuroendocrine and pharmacological received agents. Incompatibility of host immune responses to the entrapped antigens leads to an immune pathological manner instead of an immune protection which results in the disharmony of the immune effective factors. Besides the fact that stress is one of the most common effective factors in human life, it also contributed to the protection, suppression, and pathology of the immune system. In this review article, the direct and indirect effects of the stress on the function of T cells and the contributed mechanism of action will be discussed.

The new nanocomposite with various molar ratios along with magnetic properties was fabricated via precipitation (assisted by ultrasonic) procedure. The photocatalytic effects of methylene blue (∼90% degradation for optimized sample in 100 min) for finding the optimized sample performed under visible light irradiation. Moreover, the photo-antibacterial impacts of bacteria culture environments were found with an optimized sample that had effective destruction of bacteria in comparison to control group. The cytotoxicity properties of panc1 cells and magnetic behaviors of the obtained nanomaterials were evaluated and its IC50 was about 500 mg/L. As an initial step, the structural, morphological and magnetic characteristics of the fabricated nanocomposites were evaluated by Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), X-ray diffraction (XRD), energy dispersive X-ray (EDX) and MAP, UV-visible diffuse reflectance spectroscopy (DRS), and vibrating sample magnetometry (VSM) approaches. Based on SEM results, the size of nanoparticles in fabricated nanocomposite was nearly 50-70 nm for Fe 3 O 4 /SiO 2 /TiO 2 and 80-100 nm for Fe 3 O 4 /SiO 2 /TiO 2 /CeVO 4 . XRD results showed that desired nanocomposites were truly synthesized without any impurities., (Copyright © 2020 Marsooli, Rahimi-Nasrabadi, Fasihi-Ramandi, Adib, Eghbali-Arani, Ahmadi, Sohouli, Sobhani nasab, Mirhosseini, Gangali, Ehrlich and Joseph.)

Brucellosis is a worldwide bacterial zoonosis disease. Live attenuated Brucella vaccines have several drawbacks. Thus development of a safe and effective vaccine for brucellosis is a concern of many scientists. FliC protein contributes in virulence of Brucella; hence, it is a promising target for brucellosis vaccine. In this study, Mannosylated Chitosan Nanoparticles (MCN) loaded with FliC protein were synthesized as a targeted vaccine delivery system. The immunogenicity and protective efficacy of FliC and FliC-MCN against Brucella infection were evaluated in BALB/c mice. After cloning, expression and purification, FliC protein was loaded on MCN. The particle size, loading efficiency and in vitro release of the NPs were determined. Our investigation revealed that FliC and FliC-MCN could significantly increase specific IgG response (higher IgG2a titers). Besides, spleen cells from immunized mice produced high level of IFN-γ and IL-2 and low level IL-10 cytokines. Immunization with FliC and FliC-MCN conferred significant degree of protection against B. melitensis 16 M and B. abortus 544 infections. Overall these results indicate that FliC protein would be a novel potential antigen candidate for the development of a subunit vaccine against B. melitensis and B. abortus. Moreover, MCN could be used as an adjuvant and targeted vaccine delivery system., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020. Published by Elsevier B.V.)

The mucosal immune system serves as the first line of defense against Bordetella pertussis . Intranasal vaccination, due to its potential to induce systemic and mucosal immune responses, appears to prevent the initial adherence and colonization of the bacteria at the first point of contact. In the present study, two B. pertussis antigens, pertussis Toxoid (PTd) and Filamentous hemagglutinin (FHA), which play a very significant role in virulence and protection against pertussis, were encapsulate into N-trimethyl chitosan (TMC) nanoparticulate systems. After preparation of TMC nanoparticles (NPs), the NPs were characterized and their ability to induce efficient immune responses against B. pertussis was studied in a mouse model. Our findings showed that PTd + FHA-loaded TMC NPs have strong ability to induce IL-4, IL-17, IFN-γ, IgG, and IgA in the mouse model. Results from this study suggest that nasal administration of the PTd + FHA-loaded TMC NPs induced not only a systemic immune response but also a local mucosal response, which may improve the efficacy of pertussis prevention through respiratory tract transmission.

Background: Final elimination of some intracellular bacterial agents, such as Brucella, is often a complex issue and impossible to achieve, primarily due to the presence and survival of the bacteria within phagocytic cells. By penetrating into the cell membrane, drug delivery nanosystems can reduce the number of intracellular bacteria. The aim of this study was to assess the efficacy of chitosan nanoparticles on the delivery of gentamicin into Brucella infected J774A.1 murine cells in vitro., Materials and Methods: Chitosan nanoparticles (NPs) were synthesized using ionic gelation technique. The shape, size and charge of NPs, loading rate and release of the drug were investigated. Finally, the effects of gentamicin-loaded chitosan NPs (Gen-Cs) and free gentamicin on J774A.1 murine cells infected with these bacteria were examined., Results: The mean size and charge of NPs were computed as 100 nm and +28mV, respectively. The loading capacity of NPs was 22%. About 70% of the drug was released from NPs during the first 8 hours. Antimicrobial activity of the two formulations showed that MIC (minimum inhibitory concentration) of the Gen-Cs and free drug was 3.1 and 6.25 µg, respectively. The minimum bactericidal concentration of the NPs-loaded drug and free drug was 6.25 and 12.5 µg, respectively. Cell culture analysis revealed that there was a significant reduction in the load of the intercellular bacteria in J774A.1 murine cells in both formulations., Conclusion: Our results showed the Gen-Cs have a proper potential for optimal treatment of intracellular bacterial agents., (Copyright© 2019, Galen Medical Journal.)

Human epithelial growth factor receptor2 (Her2) and polymorphic epithelial mucin (MUC1) are tumor-associated antigens that have been extensively investigated in adenocarcinomas. Generally, each of these molecules was used separately for diagnosis of adenocarcinomas and as an injective vaccines in cancer therapy researches, but not in the chimeric form as an edible immunogen. In this study, Her2, MUC1, and a novel fusion structure were expressed in the seeds and hairy roots of transgenic plants appropriately. The mice groups were immunized either by feeding of transgenic seeds or hairy roots. All immunized groups showed a considerable rise in anti-glycoprotein serum IgG and IgA, and IFNɣ cytokine. However, the animals received chimeric protein showed significant higher immune responses in comparison to ones received one of these immunogen. The results indicated that the oral immunization of an animal model with transgenic plants could effectively elicit immune responses against two major tumor-associated antigens.

Fe 3 O 4 /CdWO 4 and Fe 3 O 4 /CdWO 4 /PrVO 4 magnetic nanoparticles were prepared at different molar ratios of PrVO 4 to previous layers (Fe 3 O 4 /CdWO 4 ) via the co-precipitation method assisted by a sonochemical procedure, in order to investigate the photocatalytic performance of these systems and their cytotoxicity properties. The physico-chemical properties of these magnetic nanoparticles were determined via several experimental methods: X-ray diffraction, energy dispersive X-ray spectroscopy, Fourier transformation infrared spectroscopy and ultraviolet-visible diffuse reflection spectroscopy, using a vibrating sample magnetometer and a scanning electron microscope. The average sizes of these nanoparticles were found to be in the range of 60-100 nm. The photocatalytic efficiency of the prepared nanostructures was measured by methylene blue degradation under visible light (assisted by H 2 O 2 ). The magnetic nanosystem with a 1:2:1 ratio of three oxide components showed the best performance by the degradation of ca. 70% after 120 min of exposure to visible light irradiation. Afterwards, this sample was used for the photodegradation of methyl orange, methyl violet, fenitrothion, and rhodamine-B pollutants. Finally, the mechanism of the photocatalytic reaction was examined by releasing ˙OH under UV light in a system including terephthalic acid, as well as O 2- , OH, and hole scavengers. Additionally, the cytotoxicity of each synthesized sample was assessed using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay against the human cell line PANC1 (cancer), and its IC50 was approximately 125 mg/L., Competing Interests: The authors declare no conflicts of interest.

Brucellosis is a worldwide zoonotic disease affecting domestic animals and humans. Due to several safety problems associated with live attenuated vaccines (Rev1 and RB51), it is necessary to produce an efficient and safer vaccine against Brucella. In this study, we evaluated efficacy of two novel multi-peptide vaccine candidates of FliC, 7α-HSDH, BhuA antigens with and without poly I:C adjuvant. Hence, humoral and cellular immune responses and protective efficacy were determined in immunized mice. Our investigation indicated that multi-epitope antigens showed a significant induction of Th1 immunity with high levels of specific IgG (especially the IgG2a), as well as IFN-γ and IL-2 compared to the control group. The addition of poly I:C to multi-epitope antigens improved the humoral and cellular immune responses. The multi-epitope antigens with and without poly I:C also provided cross protection against B. melitensis16M and B. abortus544 infections. The present study suggests that the novel multi-epitope vaccine candidates based on B cell, CD4 + and CD8 + T-cell epitopes of FliC, 7α-HSDH, BhuA proteins would be potential vaccine candidate against B. melitensis and B. abortus infections. Furthermore, poly I:C could be considered as a strong Th1-inducing adjuvant in designing vaccine formulation against brucellosis., (Copyright © 2019. Published by Elsevier B.V.)