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The urgency of addressing climate change, energy security, and ecological concerns has driven the global shift towards renewable energy systems. In this transformative era, green hydrogen emerges as a pivotal energy resource, promising substantial reductions in global emissions. Methods like water electrolysis, biomass gasification, and methane reforming with carbon capture and storage offer pathways to synthesize hydrogen without greenhouse gas emissions. Currently, hydrogen accounts for approximately 4% of global energy generation, with projections suggesting this could increase to 10% by 2030 and 20% by 2050. Integrating nuclear energy into hydrogen production presents a low-carbon option, promoting reliability and meeting growing demand across sectors. For instance, incorporating nuclear energy could reduce emissions by up to 90% compared to conventional methods. Categorizing hydrogen production into colors based on emissions provides a framework for understanding their environmental and socio-economic profiles. For example, green hydrogen can reduce lifecycle emissions by 60–90% compared to gray hydrogen. Life cycle analysis aids in evaluating overall impacts, guiding policy formulation and industrial decisions towards sustainability. This review examines various production pathways, highlighting their potential contributions to a carbon-neutral economy and discussing the hydrogen spectrum's role in characterizing production processes based on carbon emissions. [Display omitted] • H 2 spectrum characterizes manufacturing processes based on carbon emissions. • Renewable hydrogen reduces emissions more effectively than gray hydrogen. • Nuclear hydrogen has a low impact using Cu–Cl thermochemical cycles. • Multifaceted approaches needed to overcome hydrogen production challenges. [ABSTRACT FROM AUTHOR]

• Optimal multi-objectives energy management in EH based on probability decision-making variables. • Evs optimal platform participation in EH based on DR and energy market interactions. • Improve the two levels EH optimal operation by considering the risk index and its cost. Because the probability decision-making variables in the energy hub (EH) cause technical, economic and environmental challenges, this study presents a multi-objective planning approach based on stochastic-probability optimization algorithms. Uncertainties include the price of electricity carriers, renewable power generation, and electrical loads uncertainty (specifically electric vehicles (EVs)) based on probability distribution functions. The multi-objective optimization function includes minimizing the operation cost, minimizing the greenhouse gas emissions cost, and minimizing the EH risk cost. The stochastic-probability optimization method is programmed in both primary and secondary levels. At the primary level, the optimal operation of the EH based on the Monte Carlo method and K-means clustering is considered. To change the consumption pattern and provide a suitable platform for customer participation in optimal energy distribution, demand response (DR) programs based on energy market interactions have been modeled. At the secondary level, the EV unit optimal operation based on the Alternating direction method of multipliers (ADMM) and considering their probabilistic behavior is presented. EH risk analysis has also been developed based on the conditional value at risk (CVaR) method. The results of this study show the effectiveness of the proposed method in different scenarios and a 15.1% reduction in the total operation cost. [ABSTRACT FROM AUTHOR]

Abstract: Project: This study investigated the in vitro and in vivo effectiveness of biogenic selenium nanoparticles (Se NPs), biosynthesized by Bacillus sp. MSh-1, against Leishmania major (MRHO/IR/75/ER). Procedure: The 3-(4,5-dimethylthiozol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay was used to evaluate the cytotoxicity effects of the biogenic Se NPs against both promastigote and amastigote forms of L. major. In a separate in vivo experiment, we also determined the preventive and therapeutic effects of biogenic Se NPs in BALB/c mice following subcutaneous infected with L. major. Results: The MTT assays showed that the highest toxicity occurred after 72h against both promastigote and amastigote forms of L. major. The cytotoxicity of Se NPs was higher at all incubation times (24, 48, and 72h) against the promastigote than the amastigote form (p <0.05). The 50% inhibitory concentrations (IC50) of the Se NPs were 1.62±0.6 and 4.4±0.6μgml−1 against the promastigote and amastigote forms, respectively, after a 72-h incubation period. Apoptosis assays showed DNA fragmentation in promastigotes treated with Se NPs. In an animal challenge, prophylactic doses of biogenic Se NPs delayed the development of localized cutaneous lesions. Moreover, daily administration of Se NPs (5 or 10mgkg−1 day−1) in similarly infected BALB/c mice that had not received prophylactic doses of Se NPs also abolished the localized lesions after 14 days. Conclusion: Based on these in vitro and in vivo studies, biogenic Se NPs can be considered as a novel therapeutic agent for treatment of the localized lesions typical of cutaneous leishmaniasis. [Copyright &y& Elsevier]

Summary: Objectives: The main objective of the present study was to detect point mutations at positions 86, 184, 1034, 1042, and 1246 of the Plasmodium falciparum multidrug resistance gene (pfmdr1) in blood samples collected from malaria patients in Chabahar, a harbor city located in Southeast Iran. Methods: Twenty-six blood samples from patients infected with P. falciparum, who had a chloroquine (CQ) response failure, were collected pre-treatment. Following treatment with CQ, drug susceptibility was assessed using an in vivo test. Molecular detection of single nucleotide polymorphisms (SNPs) was carried out using the LightCycler hybridization probe assay. Results: The pfmdr1 N86Y mutation was found in six isolates (23.1%). Mutations at the four other positions were not observed in any isolates. Conclusion: The present study showed no mutation at codon positions 184, 1034, 1042, and 1246 of pfmdr1 in any of the Iranian P. falciparum isolates; thus these alleles cannot serve as markers for CQ resistance in Iran. [Copyright &y& Elsevier]

Abstract: Human infection especially with helminth parasites is an emerging health issue, as the human environment is increasingly shared with infected animals, either pets or wild life. In this survey, the intestinal content of 83 stray dogs, 22 red foxes and 10 golden Jackals collected from the West Azarbaijan, Kordestan and Kermanshah provinces in the west of Iran, were studied for the presence of helminth parasites. The percentage of different species recovered from these animals is listed as follows: From stray dogs: Toxocara canis (6.02%), Toxascaris leonina (32.53%), Ancylostoma caninum (3.61%), Oxynema sp. (1.35%), Rictularia affinis (12.05%), Taenia hydatigena (53.01%), Taenia ovis (7.23%), Taenia multiceps (4.82%), Echinococcus granulosus (13.25%), Dipylidium caninum (38.55%), Mesocestoides lineatus (26.50%) and Macracanthorhynchus hirudinaceus (4.82%). From red foxes: T. canis (4.54%), T. leonina (31.82%), A. caninum (4.54%), Uncinaria stenocephala (13.64%), Oxynema sp. (9.09%), R. affinis (54.54%), Strongyloides sp. (4.54%), Physaloptera sp. (4.54%), T. hydatigena (9.09%), E. granulosus (4.54%), D. caninum (9.09%), M. lineatus (81.82%), Joyeuxiella pasqalei (27.27%), Diplopylidium nolleri (4.54%), M. hirudinaceus (22.72%) and Macracanthorhynchus sp. (9.09%). From golden jackals: T. canis (10%), T. leonina (30%), R. affinis (50%), T. hydatigena (10%), D. caninum (20%), M. lineatus (70%), J. pasqalei (30%.), Alaria canis (10%), M. hirudinaceus (30%) and Macracanthomynchus sp. (10%). [Copyright &y& Elsevier]

Abstract: In the present study, Echinoccocus granulosus isolates collected from human, sheep and camel samples in Iran were characterized based on rostellar hook morphology of protoscoleces as well as PCR-RFLP. Morphological study on human and animal isolates showed the presence of two distinct strains of the parasite, one in sheep and the other one in camels. In this regard, rostellar hook of sheep isolates were significantly different from those of camel origin, meanwhile human isolates were found to be similar to those isolated from sheep. Molecular analysis of the ITS1 region of rDNA derived from human, sheep and camel isolates were in agreement with the morphological findings. Based on the PCR-RFLP method, the sheep and human isolates appeared to pertain to the same genotype and the camel isolates were appeared to pertain to a different genotype. [Copyright &y& Elsevier]

Glutathione S-transferase in Fasciola gigantica (FgGST) was isolated by affinity chromatography, by which highly purified enzyme was obtained. FgGST on the SDS-PAGE showed three protein bands ranging 24.5–26.5 kDa. Glutathione S-transferase (GST) activity was determined by HABIG method. FgGST was evaluated as vaccine alone or in combination with either aluminum hydroxide or saponin in sheep against F. gigantica infection. ELISA was used for detection of anti-FgGST IgG. After vaccination, all sheep were challenged with 120 metacercaria of F. gigantica. The results indicated that anti-GST IgG was not elevated after challenge. All sheep were slaughtered 24–26 weeks after challenge. The results indicated that, although after second vaccination, antibody titers rose markedly in GST–Al(OH)3 and GST–saponin groups, but declined 4 weeks after challenge. No correlation between anti-GST IgG titers and protection was observed. The highest fluke burden reduction was observed in the group vaccinated with GST–saponin (32%), but this reduction was not statistically significant in comparison with the control group. [Copyright &y& Elsevier]

In the present study, 115 stray dogs (56 males and 59 females, mixed breed), 86 golden jackal (Canis aureus, 42 males and 44 females), 60 red foxes (Vulpes vulpes, 33 males and 27 females), and three female wolves (Canis lupus) were examined for Echinococcus granulosus infection, as well as, 32,898 sheep, 10,691 goats, 15,779 cattle and 659 buffaloes for hydatid infection from five provinces in western Iran during 3 years (1997–2000). Meanwhile fertility rates of different types and forms of cysts isolated from infected animals and the viability of protoscolices were also determined. Results indicated that 19.1% of the dogs, 2.3% of the golden jackals and 5% of the red foxes were infected with Echinococcus granulosus. 11.1% of the sheep, 6.3% of the goats, 16.4% of the cattle and 12.4% of the buffaloes were also found to be infected with hydatid cyst. The cysts isolated from liver and lungs of the sheep show higher fertility rate than the cysts of liver and lungs of goats, cattle and buffaloes. [Copyright &y& Elsevier]

In the present study, excretory secretory antigens (ESA) of Toxoplasma gondii were evaluated in immunization of 8–10 week inbred female Balb/c mice. Tachyzoites of the parasite were cultured in cell-free incubation medium (RPMI-1640), and then supernatant of the medium was loaded on an ion-exchange chromatography column. Two fractions (ESA-F1 and ESA-F2) were collected from the column. For immunization of the mice, 50 were allocated into 5 groups of 10. The first, second, third, and fourth groups were immunized, twice with total-ESA, ESA-F1, ESA-F2 or toxoplasma lysate antigen (TLA), respectively. The fifth group was selected as a negative control group (non-immunized). The virulent RH strain of Toxoplasma gondii was used to challenge. Delayed-type hypersensitivity responses (DTHs) were measured by intra-footpad injection measuring induration at timed intervals. Lymphocyte transformation tests (LTTs) were done on lymph node cells using [3H] thymidine incorporation as an indication of reactivity. Peritoneal macrophages from sensitized mice were stimulated and nitric oxide was measured by Griess method. The ESA-F1 and ESA-F2 fractions were separated on poly acrylamide gel electrophoresis (PAGE) and SDS-PAGE. ESA-F1 had 4 bands on PAGE and 14 bands on SDS-PAGE. ESA-F2 had one band on PAGE and two bands on SDS-PGE. Sensitized mice showed DTH and lymphocyte transformation responses to total-ESA, ESA-F1, and ESA-F2 and peritoneal macrophages produce nitric oxide following stimulation. In challenge experiments, all non-immunized mice died within 10 days, whereas immunized mice survived for longer time periods (P<0.05). The highest survival rate was observed in mice that immunized with ESA-F2. We suggest that these antigens especially ESA-F2 should be of value for the development of new strategies for immunization against toxoplasmosis. [Copyright &y& Elsevier]

• We designed a multi-epitope DNA vaccine encoding the potential B and T-cell epitopes of T. gondii SAG1, SAG3 and SAG5 to evaluate the immune responses in BALB/c mice against acute T. gondii infection. • Immunization with multi-epitope DNA vaccine stimulated both humoral and cellular immune responses and prolonged the survival time in BALB/c mice. Co-administration of CpG-ODN adjuvant enhanced the level of protection. • The present study suggested that the DNA vaccine containing epitopes of T. gondii SAG1, SAG3 and SAG5 with CpG-ODN adjuvant might be a new model for further investigations against acute toxoplasmosis. The causative agent of toxoplasmosis, Toxoplasma gondii (T. gondii) , is able to influence the health of humans and other vertebrates. Toxoplasma may cause severe illness in the fetus and immunocompromised individuals. The high incidence and intense damages of Toxoplasma infection clearly shows the need to achieve the safe and suitable vaccine. In this study, an immunoinformatics approach was employed to design a multi-epitope DNA vaccine encoding the T. gondii SAG1, SAG3 and SAG5. The bioinformatic outputs supported the immunogenic and non-allergic natures of multi-epitope vaccine. Thereafter, the protective efficacy of the vaccine was evaluated with/without CpG-ODN adjuvant in a laboratory animal model. BALB/c mice were immunized subcutaneously with multi-epitope DNA vaccine. The in vivo findings indicated that the multi-epitope DNA vaccine elicited significant production of IgG antibodies (472.90 ± 2.74 ng/ml) as well as IFN-γ (173.71 ± 26.39 pg/ml) (p < 0.001). Moreover, a significant reduced parasite-burden (17,470 per mg of spleen) and prolonged survival time (9 days) were observed in the immunized groups compared to the controls (p < 0.05). The low values of IL-4 (22.5 ± 0.16 pg/ml) were detected in vaccinated mice compared to the control (PBS) (p > 0.05). In addition, CpG-ODN as an adjuvant increased the immune efficacy of the multi-epitope DNA vaccine. In multi-epitope vaccine+CpG-ODN group, the values of IgG antibodies (535.90 ±7.29 ng/ml) and IFN-γ (358.21 ± 32.70 pg/ml) were significanly higher than the multi-epitope vaccine group. Meanwhile, an increased survival time (10 days) and fewer parasite load (15,485 per mg of spleen) were observed in multi-epitope vaccine+CpG-ODN group. The results revealed that the DNA vaccine containing epitopes of SAG1, SAG3 and SAG5 adjuvanted with CpG-ODN might be a new model for further investigations against acute T. gondii infection. [ABSTRACT FROM AUTHOR]

Abstract Rhoptry proteins (ROPs) are involved in the different stages of Toxoplasma gondii (T. gondii) invasion and are also critical for survival within host cells. ROP8 is expressed in the early stages of infection and have a key role in the parasitophorous vacuole (PV) formation. In this paper, we have combined several bioinformatics online servers for immunogenicity prediction of ROP8 protein. In this study, several bioinformatics approaches were used to analyze the different aspects of ROP8 protein, including the physico-chemical properties, transmembrane domain, subcellular localization, secondary and tertiary structure, B and T-cell potential epitopes, and other important characteristics of this protein. The findings showed that ROP8 protein had 60 potential post-translational modification sites. Also, only one transmembrane domain was recognized for this protein. The secondary structure of ROP8 protein comprises 33.04% alpha-helix, 18.26% extended strand, and 48.70% random coil. Moreover, several potential B and T-cell epitopes were identified for ROP8. In addition, the obtained findings from antigenicity and allergenicity evaluation remarked that this protein is immunogenic and non-allergen. Based on the results of Ramachandran plot, 94.8%, 4.1%, and 1.1% of amino acid residues were incorporated in the favored, allowed, and outlier regions, respectively. This paper provides a foundation for further investigations, and laid a theoretical basis for the development of an appropriate vaccine against toxoplasmosis. More studies are needed experimentally using the ROP8 alone or in combination with other antigens in the future. Highlights • Toxoplasma gondii can infect a broad range of warm-blooded vertebrate species worldwide • ROP8 is expressed in bradyzoites and tachyzoites stages of T. gondii. It can be expressed in the early stages of infection and have a key role in parasitophorous vacuole formation. • We have combined several bioinformatics online servers for immunogenicity prediction of ROP8 protein. • Bioinformatics online servers revealed that ROP8 protein had several excellent B and T-cells epitopes, suggesting that it would be become an outstanding vaccine target. • This paper provides a foundation for further investigations, and laid a theoretical basis for the development of an appropriate vaccine against toxoplasmosis. [ABSTRACT FROM AUTHOR]

Neospora caninum and Toxoplasma gondii are two closely related protozoan parasites that have been detected from various species of bird hosts. However, little is known about the prevalence of N. caninum and T. gondii in crows. Hence, we examined the molecular frequency of N. caninum and T. gondii in the brain samples of hooded crows ( Corvus cornix ) that collected from different public parks of Tehran, Iran by nested-PCR method. We used the primers targeting the Nc 5 and GRA 6 genes for detection of N. caninum and T. gondii , respectively. From a total of 55 brain samples, 5 (9.9%) and 9 (16.36%) samples were positive for N. caninum and T. gondii , respectively. Sequencing of a N. caninum isolate revealed 95%–100% identity with the deposited N. caninum in GenBank. Genotyping of T. gondii isolates by PCR-RFLP analysis of the GRA6 gene revealed type III genotype in 8 isolates. The results of this study indicate that hooded crows may have a putative role in transmission of N. caninum and T. gondii to canines and felines definitive hosts, respectively. [ABSTRACT FROM AUTHOR]

This study was designed to evaluate the safety, immunogenicity, and efficacy of a single dose of L. infantum (LiCen -/- ) live attenuated candidate vaccine against canine leishmaniasis (CanL). Eighteen healthy domestic dogs with no anti- Leishmania antibodies and negative leishmanin skin test (LST) were randomly inoculated intravenously with either L. infantum (LiCen -/- ) vaccine candidate in 10 dogs or phosphate-buffered saline (PBS) in 8 dogs. The safety, immunogenicity, and efficacy rate of L. infantum (LiCen -/- ) vaccine candidate against CanL were evaluated by different criteria, including clinical manifestations, injection-site lesion, hematology and biochemistry values, anti- Leishmania antibodies using direct agglutination test (DAT), delayed-type hypersensitivity (DTH) using LST, and CD4+ and CD8+ T-cells subsets, as well as by measuring interferon (IFN-γ), interleukin (IL-23), IL-17, and IL-10 cytokines. Spleen aspiration and detection of Leishmania parasite using parasitological examinations (microscopy and culture) were performed in both vaccinated and control groups. Two months after intervention, each dog was challenged intraperitoneally (IP) with wide type (WT) L. infantum. Two-month follow-up post vaccination showed no clinical signs and serious side effects associated with the vaccination. A significant increase was found in the expression of IL-17, CD4+, and CD8+ gene transcripts in PBMCs, as well as increased levels of Th1 cytokines, and reduction of Th2 cytokine. The efficacy of the vaccine candidate was calculated to be 42.85%. While the time window for assessing the vaccine's effectiveness was too limited to draw any real conclusions but the preliminary results showed a moderate efficacy rate due to inoculation a single dose of L. infantum (LiCen -/- ) vaccine candidate. Further investigations with more sample sizes and multiple doses of the vaccine candidate using natural challenges in the endemic areas of CanL are recommended. • A single dose of LiCen -/- mutant showed to be safe with efficacy rate of 42.85%. • Significant increase was observed in the expression of IL-17, CD4 + and CD8 + gene transcripts in PBMCs of LiCen -/- in vaccinated dogs compared to controls. • Increased levels of Th1 cytokines, and reduction of Th2 cytokines, in the vaccinated group acompared to control group. [ABSTRACT FROM AUTHOR]

Abstract: Toxoplasma gondii, the pathogen of toxoplasmosis, can infect most mammals and birds. The high incidence and severe or lethal damages of toxoplasmosis clearly indicate the need for the development of a more effective vaccine. We constructed a DNA cocktail, containing plasmids encoding the full-length SAG1 and ROP2 genes of T. gondii and evaluated its immune response and protective efficacy in comparison with single-gene vaccines and control groups. We immunized BALB/c mice intramuscularly three times. DNA cocktail elicited IgG and IFN-γ, TNF-α and IL-2 greater than single-gene plasmids and increased survival time against a lethal challenge with the highly virulent T. gondii RH strain. The current study shows that pc-SAG1+ pc-ROP2 as a cocktail DNA vaccine produces higher Th1 immune response than single-gene plasmids and cocktail DNA is effective to prime an enhanced and balanced specific immunity. [ABSTRACT FROM AUTHOR]

• Toxoplasma gondii (T. gondii) is an obligate intracellular parasite which is adapted to a wide spectrum of warm-blooded vertebrates. • ROP8 is critical for acute virulence of T. gondii and is expressed in tachyzoite and bradyzoite stages. • We designed a novel multi-epitope DNA vaccine from ROP8 protein to evaluate the immunity and protective efficacy against T. gondii infection. • The findings revealed that the multi-epitope ROP8 DNA vaccine induced strong humoral and cellular responses and prolonged the survival time in BALB/c mice. Rhoptry proteins (ROPs) are involved in the cell invasion and parasitophorous vacuole (PV) formation and also vital for survival of Toxoplasma gondii (T. gondii) within host cells. ROP8 have a main role during the early phase of infection and can express in tachyzoite and bradyzoite forms. In the present study, we designed a novel multi-epitope DNA vaccine encoding the potential B and T-cell epitopes from ROP8 protein to evaluate the immunity and protective efficacy against acute T. gondii infection in BALB/c mice. For this purpose, several bioinformatics online servers were used. At first, the potential epitopes were selected for T and B cells using immune epitope database (IEDB) and BCPREDS online services. Then, the selected epitopes were fused together by SAPGTP linker. Finally, the physico-chemical features, secondary and tertiary structures, antigenicity, and allergenicity of the peptide were evaluated through different bioinformatics tools. Lastly, the multi-epitope peptide was successfully cloned into pcDNA3.1 expression vector. The DNA vaccine was subcutaneously injected into BALB/c mice and the immune responses of the vaccinated mice and controls were determined. The obtained results revealed that the multi-epitope ROP8 peptide has 183 amino acid residues with average molecular weight (MW) of 18.974 kDa. More than 98 % residues of the peptide were incorporated in favored and allowed regions of the Ramachandran plot. The antigenicity of multi-epitope peptide were estimated 0.8751 and 0.7649 by ANTIGENpro and VaxiJen servers, respectively. BALB/c mice immunized with DNA vaccine showed significantly increased the level of specific anti- T. gondii antibodies (P < 0.05), and a mixed IgG1/IgG2a response with predominance of IgG2a production. The immunized mice also displayed a TH1-type cellular immune response with production of IFN-γ and prolonged survival time, compared with the control groups (P < 0.05). The findings revealed that the multi-epitope ROP8 DNA vaccine induced strong humoral and cellular responses and prolonged the survival time in BALB/c mice, suggesting selection of potential epitopes may be a promising strategy for the design of multi-epitope-based vaccines. [ABSTRACT FROM AUTHOR]