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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.
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We report the effect of a rodent control program on the incidence of zoonotic cutaneous leishmaniasis in an endemic region of Iran. A 1-year interruption in rodent control led to 2 years of increased incidence of zoonotic cutaneous leishmaniasis. Restarting rodent control led to a decline of zoonotic cutaneous leishmaniasis.
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Leishmaniasis Cutánea , Zoonosis , Irán/epidemiología , Leishmaniasis Cutánea/epidemiología , Leishmaniasis Cutánea/prevención & control , Animales , Zoonosis/epidemiología , Zoonosis/prevención & control , Humanos , Incidencia , Control de Roedores/métodos , Roedores/parasitología , Reservorios de Enfermedades/parasitología , Reservorios de Enfermedades/veterinariaRESUMEN
BACKGROUND: Klebsiella is an opportunistic pathogen, which is the most common causes of nosocomial infections. To date, the prevalence of ESBL-producing pathogens has increased and is associated with mortality, morbidity, and healthcare costs. The aim of this investigation was to determine the frequency of blaSHV, blaTEM, and blaCTX-M genes from Klebsiella pneumonia isolated from patients with UTI in the city of Qom. METHODS: In the cross-sectional study, a total of 500 urinary samples were cultured in MacConkey agar and identified using the biochemical test. For a total of 340 positive K. pneumonia samples the antimicrobial susceptibility was determined using the Kirby-Bauer disc diffusion approach. For molecular genotyping, the frequencies of blaSHV, blaCTX-M, and blaTEM genes were determined using a polymerase chain reaction (PCR) method. RESULTS: Our finding revealed that a total of 340 K. pneumonia isolates 110 isolates (32.35%) were ESBL producers by the phenotypic method. All of these isolates were assessed by PCR for blaSHV, blaCTX-M, and blaTEM genes. The PCR results demonstrated that the frequencies of blaTEM, blaCTX-M, and blaSHV genes were 59.09% (65 isolates), 74.54% (82 isolates), and 74.54% (82 isolates), respectively. CONCLUSIONS: According to our findings, with the higher prevalence of ESBL-producing isolates in the clinical, early detection, and follow-up procedures are critical strategies to the prevention of the spread of multidrug resistant isolates.
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Klebsiella pneumoniae , Neumonía , Humanos , Klebsiella pneumoniae/genética , beta-Lactamasas/genética , Estudios Transversales , Antibacterianos/uso terapéutico , Neumonía/tratamiento farmacológicoRESUMEN
Microalgae are powerful source for nutritionally valuable components as proteins, carbohydrates and especially unsaturated fatty acids. Microalgae may be employed in pharmaceutical, food, cosmetic, health industries, and biofuels. In this study for looking at high-level unsaturated fatty acids species, from 31 strains, by comparing growth curves, the best strain with a high growth rate and lipid content was selected by red Nile staining. It was determined by molecular identification that this strain belongs to the genus Chlorella sp. and is deposited into the Agricultural Biotechnology Research Institute of Iran Culture collection with culture collection number ABRIICC 30,041. Biomass analysis after growth optimization by response surface methodology showed that the selected strain had a specific growth rate of 0.216 ± 0.008 d-1, biomass productivity of 142.58 ± 4.41 mg/Ld, and lipid content of 13.9 ± 0.26% with a high level of unsaturated fatty acids of 53.15%. It also included 51.3 ± 0.53% protein with a very high quality essential amino acids of 40.36%, the most lysine (8.77%) and arginine (13.31%) has been reported until now, and 26.9 ± 0.23% carbohydrates in photoautotroph condition. By MTT assay, there is no effect of cytotoxicity. This research introduces a potent native strain comparable with commercial strains that can be a hopeful source for food supplements and valuable bioactive ingredients in functional foods.
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Chlorella , Microalgas , Ácidos Grasos/análisis , Lisina/metabolismo , Microalgas/metabolismo , Arginina/metabolismo , Ácidos Grasos Insaturados/metabolismo , Carbohidratos , Proteínas/metabolismo , Suplementos Dietéticos/análisis , Biomasa , BiocombustiblesRESUMEN
BACKGROUND: Melanoma differentiation-associated gene 7 (Mda-7) encodes IL-24, which can induce apoptosis in cancer cells. A novel gene therapy approach to treat deadly brain tumors, recombinant mda-7 adenovirus (Ad/mda-7) efficiently kills glioma cells. In this study, we investigated the factors affecting cell survival and apoptosis and autophagy mechanisms that destroy glioma cells by Ad/IL-24. METHODS: Human glioblastoma U87 cell line was exposed to a multiplicity of infections of Ad/IL-24. Antitumor activities of Ad/IL-24 were assessed by cell proliferation (MTT) and lactate dehydrogenase (LDH) release analysis. Using flow cytometry, cell cycle arrest and apoptosis were investigated. Using the ELISA method, the tumor necrosis factor (TNF-α) level was determined as an apoptosis-promoting factor and Survivin level as an anti-apoptotic factor. The expression levels of TNF-related apoptosis inducing ligand(TRAIL) and P38 MAPK genes were assessed by the Reverse transcription-quantitative polymerase chain reaction(RTqPCR) method. The expression levels of caspase-3 and protein light chain 3-II (LC3-II) proteins were analyzed by flow cytometry as intervening factors in the processes of apoptosis and autophagy in the cell death signaling pathway, respectively. RESULTS: The present findings demonstrated that transduction of IL-24 inhibited cell proliferation and induced cell cycle arrest and cell apoptosis in glioblastoma. Compared with cells of the control groups, Ad/IL24-infected U87 cells exhibited significantly increased elevated caspase-3, and TNF-α levels, while the survivin expression was decreased. TRAIL was shown to be upregulated in tumor cells after Ad/IL-24 infection and studies of the apoptotic cascade regulators indicate that Ad/IL-24 could further enhance the activation of apoptosis through the TNF family of death receptors. In the current study, we demonstrate that P38 MAPK is significantly activated by IL-24 expression. In addition, the overexpression of mda-7/IL-24 in GBM cells induced autophagy, which was triggered by the upregulation of LC3-II. CONCLUSIONS: Our study demonstrates the antitumor effect of IL-24 on glioblastoma and may be a promising therapeutic approach for GBM cancer gene therapy.
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Glioblastoma , Humanos , Survivin/genética , Glioblastoma/patología , Caspasa 3/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo , Regulación hacia Arriba , Factor de Necrosis Tumoral alfa/metabolismo , Apoptosis , Proteínas Reguladoras de la Apoptosis/metabolismo , Autofagia/genética , Línea Celular Tumoral , Ligando Inductor de Apoptosis Relacionado con TNF/farmacologíaRESUMEN
Biofilm formation and resistance to antibiotics in pathogenic bacteria are important concerns in the treatment of infectious diseases. A new rapid, eco-friendly and cost-effective strategy to overcome these problems is the use of microbial exopolysaccharides (EPS) for green synthesis of various metal nanoparticles (NPs). This study used EPS from a native probiotic Lactobacillus isolate to synthesize silver nanoparticles (AgNPs) with effective antimicrobial, antibiofilm and antioxidant properties. AgNPs were synthesized by 10 mg of EPS of Lactobacillus paracasei (L. paracasei MN809528) isolated from a local yogurt. The characteristics of EPS AgNPs were confirmed using UV-VIS, FT-IR, DLS, XRD, EDX, FE-SEM, and zeta potential. Antimicrobial, antibiofilm and antioxidant activities of EPS AgNPs were evaluated by the agar well diffusion, microtiter dilution, SEM electron microscopy, and DPPH radical absorption methods, respectively. Spectroscopy data indicated the presence of a 466-nm peak as a feature of AgNPs. FT-IR confirmed the presence of biological agents in the synthesis of AgNPs. FE-SEM results showed that the synthesized AgNPs had a spherical shape with the size of 33-38 nm. Synthesized AgNPs at a concentration of 100 mg/ml demonstrated a significant inhibitory activity compared to chemically synthesized AgNPs. These NPs, exhibited the greatest effect of inhibiting the Escherichia coli and Pseudomonas aeruginosa biofilm formation at sub-MIC concentration, and the best effect of DPPH radical as antioxidant activity was determined at 50-µg/ml concentration. Our findings reveal that EPS AgNPs synthesized by the native isolate of L. paracasei (MN809528) is an inexpensive and environment-friendly candidate for application in pharmaceuticals fields.
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Antiinfecciosos , Lacticaseibacillus paracasei , Nanopartículas del Metal , Antioxidantes/farmacología , Antioxidantes/química , Plata/farmacología , Plata/química , Espectroscopía Infrarroja por Transformada de Fourier , Nanopartículas del Metal/química , Pruebas de Sensibilidad Microbiana , Tamaño de la Partícula , Antiinfecciosos/farmacología , Antibacterianos/farmacología , Antibacterianos/química , Extractos Vegetales/farmacología , Escherichia coli , BiopelículasRESUMEN
The halophilic microorganisms living in extreme environments contain high concentrations of carotenoids with notable medical abilities. The purpose of this study was to evaluate the anticancer effect of carotenoids extracted from native Iranian halophilic microorganisms with the ability to inhibit breast cancer cell line. To begin the study, 40 halophilic strains were cultured, and 8 strains capable of producing pigmented colonies were chosen from those cultured strains. In the next step, from among 8 strains using MTT assay, 1 capable of reducing cell viability of the breast cancer MCF-7 cell line was chosen as a selective strain. The principal carotenoid was characterized using UV-visible, FT-IR spectroscopic, and LC-MASS analyses. Using real time PCR technique, the expression of genes specific for apoptosis, in the presence or absence of carotenoid, was examined. Among all strains, carotenoid extracted from strain A15 had the most potent cytotoxic effect on breast cancer cell line (IC50 = 0.0645 mg/mL). 16S rRNA gene analysis showed that strain A15 had similarity with Haloarcula hispanica for about 99.5%. According to the analysis results, it could be estimated that the principal carotenoid extracted form Haloarcula sp. A15 was similar to bacterioruberin. Both early and late apoptosis were increased significantly about 10% and 39%, respectively, due to upregulation of CASP3, CASP8, BAX genes expression in MCF-7 cell line. In contrast, the expression of genes MKI67, SOX2 were significantly downregulated in treated MCF-7 cell line. The results of this study showed that Halophilic archaeon strain could be a good candidate for the production of high added-value bacterioruberin due to its possible anticancer properties.
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Neoplasias de la Mama , Haloarcula , Humanos , Femenino , Haloarcula/genética , Haloarcula/metabolismo , Espectroscopía Infrarroja por Transformada de Fourier , ARN Ribosómico 16S/genética , Neoplasias de la Mama/tratamiento farmacológico , Irán , Carotenoides/farmacología , Carotenoides/química , Carotenoides/metabolismoRESUMEN
Background: The presence of microbiome in the blood samples of healthy individuals has been addressed. However, no information can be found on the healthy human blood microbiome of Iranian subjects. The current study is thus aimed to investigate the existence of bacteria or bacterial DNA in healthy individuals. Methods: Blood samples of healthy subjects were incubated in BHI broth at 37 °C for 72 h. The 16S rRNA PCR and sequencing were performed to analyze bacterial isolates. The 16S rRNA PCR was directly carried out on DNA samples extracted from the blood of healthy individuals. Next generation sequencing (NGS) was conducted on blood samples with culture-positive results. Results: Fifty blood samples were tested, and six samples were positive by culture as confirmed by Gram staining and microscopy. The obtained 16S rRNA sequences of cultured bacterial isolates revealed the presence of Bacilli and Staphylococcus species by clustering in the GeneBank database (≥97% identity). The 16S rRNA gene sequencing results of one non-cultured blood specimen showed the presence of Burkholderia. NGS results illustrated the presence of Romboutsia, Lactobacillus, Streptococcus, Bacteroides, and Staphylococcus in the blood samples of positive cultures. Conclusion: The dormant blood microbiome of healthy individuals may give the idea that the steady transfer of bacteria into the blood does not necessarily lead to sepsis. However, the origins and identities of blood-associated bacterial rDNA sequences need more evaluation in the healthy population.
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Bacterias , Bacterias/genética , ADN Bacteriano/genética , ADN Ribosómico/genética , Voluntarios Sanos , Humanos , Irán , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN/métodosRESUMEN
Aim: Phage therapy, as an effective and specific method in the treatment of multidrug-resistant (MDR) bacterial infections, has attracted the attention of many researchers. Methods and results: In this study, a double-stranded DNA phage with the ability of lysing some strains of MDR Klebsiella pneumoniae (vB_Kpn_3) was isolated from hospitals' wastewater and then characterized morphologically and genetically. Transmission electron microscopy and genetic analyses have revealed that vB_Kpn_3 is a member of Siphoviridae family. One-step growth curve also showed a burst time of 35 min and a burst size of 31 PFU/ml. The genome of the phage is composed of 112,080 bp with 41.33% G + C content carrying 186 open reading frames. Conclusion: vB_Kpn_3 is a broad host range phage that infects MDR K. pneumoniae and some other species of Enterobacteriaceae such as Escherichia coli and Salmonella typhi. In addition, no antibiotic resistance and toxin genes were detected in its genome.
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Bacteriófagos , Terapia de Fagos , Bacteriófagos/genética , Genoma Viral , Klebsiella pneumoniae , FilogeniaRESUMEN
Legionella pneumophila (L. pneumophila) is one of the main pathogens, causing pneumonia and respiratory tract infections, especially in patients with ventilator-associated pneumonia (VAP). This study aimed to approve the hypothesis that the serogroup distribution of L. pneumophila isolates from patients is correlated with Legionella strains in the environment. A total of 280 bronchoalveolar lavage (BAL) samples from VAP patients admitted to the intensive care unit (ICU) as well as 116 water samples from different sources in four hospitals in Tehran, Iran, were evaluated for the presence of L. pneumophila infection by culture, nested polymerase chain reaction (PCR), real-time PCR, and sequencing for genetic diversity. The molecular and culture methods found 24 (8.6%) and 5 (1.8%) samples to be positive for L. pneumophila in VAP patients, while they found 23 (19.8%) and 8 (6.9%) positive samples in water resources, respectively. The sequencing results indicated that all positive clinical samples and 14 (60.8%) environmental samples were belonged to L. pneumophila serogroup 1. Smoking, age, length of ICU stay, and duration of ventilator use had strong relationship with L. pneumophila infectivity. In conclusion, this is the first report from Iran to determine minor differences in the serogroup distribution of environmental and clinical strains. However, further studies are needed to confirm this relationship in different regions of Iran.
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Legionella pneumophila , Legionella , Enfermedad de los Legionarios , Neumonía Asociada al Ventilador , Hospitales , Humanos , Irán , Enfermedad de los Legionarios/epidemiología , Neumonía Asociada al Ventilador/epidemiología , Reacción en Cadena en Tiempo Real de la Polimerasa , Serogrupo , Agua , Microbiología del AguaRESUMEN
INTRODUCTION: Asthma is becoming a major health problem in many countries. Immune responses in allergic asthma, as the most prevalent asthmatic phenotype, are mediated mostly by a subtype of T lymphocytes referred to as the effector lineage of Type 2 Th cells (Th2). The development of Th2 cells is mainly governed by a zinc finger transcription factor, i.e., GATA-binding protein 3 (GATA3). Allergic asthma is a complex disease, and vitamin D deficiency has been named as a non-genetic risk factor for its development. Vitamin D, a steroid hormone belonging to the family of nuclear receptors, has shown significant immunosuppressive effects in previous studies. In this study, given its immunomodulatory properties, we aimed to investigate the effects of different concentrations of vitamin D on GATA3 gene expression in peripheral blood mononuclear cells (PBMCs), including Th2 cells, and compare GATA3 expression levels between PBMCs taken from allergic asthmatic patients and healthy controls. RESULTS: The total sample size was 40 and the quantitative real-time PCR (qPCR) procedure was applied to assess the mRNA expression levels of GATA3 in different groups. Collectively, our results demonstrated that the expression of GATA3 in PBMCs taken from patients with allergic asthma is lower than in that from healthy controls. In addition, in the control group, cells co-cultured with vitamin D had a significantly increased GATA3 expression. However, in the patient group, such an increase was only observed in cells treated with 10â»7M-vitamin D. By contrast, incubation with vitamin D at the concentration of 10-6 M slightly decreased the expression of GATA3 among patients. CONCLUSION: In summary, it is likely that vitamin D should regulate GATA3 gene expression in the PBMCs in a dose-dependent manner. The impacts of this steroid hormone can also differ between the status of health and allergic asthma in either extent or direction.
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The application of biological nanoparticles (NPs) can be considered as a way to overcome the problem of antifungal resistance in pathogenic fungi. This study takes a new approach to biosynthesized NPs influence on the expression of CYP51A and HSP90 antifungal resistance genes in Aspergillus fumigatus and A. flavus, and comparison with antifungal agents. Selenium NPs (Se-NPs) were biosynthesized using Aspergillus strains and their production was proved by several methods including, UV-Vis, XRD, FTIR, FESEM, and EDX techniques. The minimum inhibitory concentrations (MICs) of Aspergillus strains were determined using the CLSI M38-A2 broth microdilution method. The differences in expression levels of CYP51A and HSP90 genes were examined between untreated and treated of A. fumigatus and A. flavus using itraconazole and amphotericin B and biosynthesized Se-NPs through real-time PCR. After confirming the results of NPs synthesis, the MIC of itraconazole and amphotericin B against A. fumigatus and A. flavus was 4 µg/ml. Based on the real-time PCR results, the obtained ∆∆CTs for these strains were -0.18, -1.46, and -1.14. Whereas the MIC values for treated samples with Se-NPs have decreased to 0.5 µg/ml, and the ∆∆CTs for these were -0.25, -1.76, and -1.68. The expression of CYP51A and HSP90 genes was significantly down-regulated through the use of Se-NPs against A. fumigatus and A. flavus.
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Nanopartículas , Selenio , Anfotericina B/farmacología , Antifúngicos/farmacología , Aspergillus/genética , Aspergillus flavus , Aspergillus fumigatus/genética , Itraconazol/farmacología , Pruebas de Sensibilidad Microbiana , Selenio/farmacología , Triazoles/farmacología , Voriconazol/farmacologíaRESUMEN
AIMS: The use of cyanobacterial cell extracts for the synthesis of zinc oxide nanoparticles (ZnO NPs) seems to be superior to other methods of synthesis because of its a green, environmentally friendly and low-cost approach. In this study, the cell extract of a newly characterized cyanobacterial strain Desertifilum sp. EAZ03 was used for the biosynthesis of ZnO NPs. The antimicrobial, antibiofilm and anticancer activities of the biosynthesized ZnO NPs (hereinafter referred to as CED-ZnO NPs) were examined as well. METHODS AND RESULTS: UV-Vis spectroscopy analysis of CED-ZnO NPs showed an absorbance band at 364 nm, and powder X-ray diffraction analysis confirmed the purity of the synthesized nanoparticles. The analyses of scanning electron microscopy and transmission electron microscopy images revealed that CED-ZnO NPs were rod-shaped with a size of 88 nm. The study of the biological features of CED-ZnO NPs showed a significant antimicrobial potential against the bacterial strains tested. CED-ZnO NPs were able to impede the biofilm formation by Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa up to 80%, 89% and 85%, respectively. The nanoparticles also showed 69%, 70% and 62% degrading activity against S. aureus, E. coli and P. aeruginosa 1-day-old biofilms, respectively. The antibiofilm activity of the synthesized nanoparticles was investigated by confocal laser scanning microscopy. The MTT assay showed that CED-ZnO NPs, at a concentration of 100 µg/ml, had less cytotoxicity towards normal lung (MRC-5) cells, at the half, compared to cancerous lung alveolar epithelial (A549) cells. The minimum inhibitory concentration and minimum bactericidal concentration values of CED-ZnO NPs against E. coli, P. aeruginosa and S. aureus were 1500, 2000 and 32 µg/ml, and 2500, 3500 and 64 µg/ml, respectively. CONCLUSIONS: The multifunctional CED-ZnO NPs seem to be promising for possible applications in the therapeutic and pharmaceutical industries. SIGNIFICANCE AND IMPACT OF THE STUDY: This study proposes a new approach for the biosynthesis of zinc oxide nanoparticles using a newly characterized cyanobacterial strain Desertifilum sp. EAZ03. The considerable antimicrobial, antibiofilm and anticancer activities of the biosynthesized zinc oxide nanoparticles further emphasize the emerging role of microbial systems in the green synthesis of metal oxide nanoparticles.
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Cianobacterias , Nanopartículas del Metal , Óxido de Zinc , Antibacterianos/farmacología , Antineoplásicos/farmacología , Biopelículas/efectos de los fármacos , Extractos Celulares , Escherichia coli , Tecnología Química Verde , Pruebas de Sensibilidad Microbiana , Extractos Vegetales/farmacología , Staphylococcus aureus , Óxido de Zinc/farmacologíaRESUMEN
BACKGROUND AND OBJECTIVES: Multi-drug-resistant Enterobacter aerogenes is associated with various infectious diseases that cannot be easily treated by antibiotics. However, bacteriophages have potential therapeutic applications in the control of multi-drug-resistant bacteria. In this study, we aimed to isolate and characterize of a lytic bacteriophage that can lyse specifically the multi-drug-resistant (MDR) E. aerogenes. MATERIALS AND METHODS: Lytic bacteriophage was isolated from Qaem hospital wastewater and characterized morphologically and genetically. Next-generation sequencing was used to complete genome analysis of the isolated bacteriophage. RESULTS: Based on the transmission electron microscopy feature, the isolated bacteriophage (vB-Ea-5) belongs to the family Myoviridae. vB-Ea-5 had a latent period of 25 minutes, a burst size of 13 PFU/ml, and a burst time of 40 min. Genome sequencing revealed that vB-Ea-5 has a 135324 bp genome with 41.41% GC content. The vB-Ea-5 genome codes 212 ORFs 90 of which were categorized into several functional classes such as DNA replication and modification, transcriptional regulation, packaging, structural proteins, and a host lysis protein (Holin). No antibiotic resistance and toxin genes were detected in the genome. SDS-PAGE of vB-Ea-5 proteins exhibited three major and four minor bands with a molecular weight ranging from 18 to 50 kD. CONCLUSION: Our study suggests vB-Ea-5 as a potential candidate for phage therapy against MDR E. aerogenes infections.
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The use of gamma-irradiated influenza A virus (γ-Flu), retains most of the viral structural antigens, represent a promising option for vaccine development. However, despite the high effectiveness of γ-Flu vaccines, the need to incorporate an adjuvant to improve vaccine-mediated protection seems inevitable. Here, we examined the protective efficacy of an intranasal gamma-irradiated HIN1 vaccine co-administered with a plasmid encoding mouse interleukin-28B (mIL-28B) as a novel adjuvant in BALB/c mice. Animals were immunized intranasally three times at one-week intervals with γ-Flu, alone or in combination with the mIL-28B adjuvant, followed by viral challenge with a high lethal dose (10 LD50) of A/PR/8/34 (H1N1) influenza virus. Virus-specific antibody, cellular and mucosal responses, and the balance of cytokines in the spleen IFN-γ, IL-12, and IL-4) and in lung homogenates (IL-6 and IL-10) were measured by ELISA. The lymphoproliferative activity of restimulated spleen cells was also determined by MTT assay. Furthermore, virus production in the lungs of infected mice was estimated using the Madin-Darby canine kidney (MDCK)/hemagglutination assay (HA). Our data showed that intranasal immunization with adjuvanted γ-Flu vaccine efficiently promoted humoral, cellular, and mucosal immune responses and efficiently decreased lung virus titers, all of which are associated with protection against challenge. This combination also reduced IL-6 and IL-10 levels in lung homogenates. The results suggest that IL-28B can enhance the ability of the vaccine to elicit virus-specific immune responses and could potentially be used as an effective adjuvant.
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Adyuvantes Inmunológicos/administración & dosificación , Citocinas/inmunología , Inmunidad Celular/inmunología , Inmunidad Mucosa/inmunología , Subtipo H1N1 del Virus de la Influenza A/inmunología , Infecciones por Orthomyxoviridae/inmunología , Administración Intranasal/métodos , Animales , Anticuerpos Antivirales/inmunología , Antígenos Virales/inmunología , Perros , Femenino , Inmunización/métodos , Vacunas contra la Influenza/inmunología , Pulmón/inmunología , Pulmón/virología , Células de Riñón Canino Madin Darby , Ratones , Ratones Endogámicos BALB C , Infecciones por Orthomyxoviridae/virología , Vacunación/métodosRESUMEN
Despite tremendous efforts toward vaccination, influenza remains an ongoing global threat. The induction of strain-specific neutralizing antibody responses is a common phenomenon during vaccination with the current inactivated influenza vaccines, so the protective effect of these vaccines is mostly strain-specific. There is an essential need for the development of next-generation vaccines, with a broad range of immunogenicity against antigenically drifted or shifted influenza viruses. Here, we evaluate the potential of whole inactivated vaccines, based on chemical and physical methods, as well as new approaches to generate cross-protective immune responses. We also consider the mechanisms by which some of these vaccines may induce CD8+ T-cells cross-reactivity with different strains of influenza. In this review, we have focused on conventional and novel methods for production of whole inactivated influenza vaccine. As well as chemical modification, using formaldehyde or ß-propiolactone and physical manipulation by ultraviolet radiation or gamma-irradiation, novel approaches, including visible ultrashort pulsed laser, and low-energy electron irradiation are discussed. These two latter methods are considered to be attractive approaches to design more sophisticated vaccines, due to their ability to maintain most of the viral antigenic properties during inactivation and potential to produce cross-protective immunity. However, further studies are needed to validate them before they can replace traditional methods for vaccine manufacturing.
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Virus de la Influenza A/inmunología , Virus de la Influenza B/inmunología , Vacunas contra la Influenza/biosíntesis , Vacunas contra la Influenza/inmunología , Vacunas de Productos Inactivados/biosíntesis , Vacunas de Productos Inactivados/inmunología , Vacunología/métodos , Animales , Humanos , Gripe Humana/prevención & control , Infecciones por Orthomyxoviridae/prevención & controlRESUMEN
Cyanobacteria, as one of the largest groups of phototrophic bacteria, have a high potential as an excellent source of fine chemicals and bioactive compounds, including lipid-like compounds, amino acid derivatives, proteins, and pigments. This study aimed to synthesize ZnO nanoparticles using the cell extract of the cyanobacterium Nostoc sp. EA03 (CEN-ZnO NPs) through a rapid and eco-friendly approach. The biosynthesized nanoparticles, CEN-ZnO NPs, were characterized by UV-Vis spectroscopy, X-ray diffraction (XRD), zeta potential measurement, differential scanning calorimetry (DSC)/thermogravimetric analysis (TGA), FTIR, SEM, TEM, and EDX spectroscopy. The UV-Vis spectrum showed an absorption peak at 370 nm. The star-shaped CEN-ZnO NPs, as observed in the TEM and SEM images, had an average diameter of 50-80 nm. MIC and MBC values for E. coli, P. aeruginosa and S. aureus, were determined to be, respectively, 2000, 2000, and 64 µg ml-1, and 2500, 2500 and 128 µg ml-1. Further analysis through confocal laser scanning microscopy (CLSM) provided the observable confirmation that the CEN-ZnO NPs stunted the bacterial growth, preventing the formation of exopolysaccharides. The AFM analysis of surface topography of bacterial biofilm samples treated with CEN-ZnO NPs showed a rugged topography in some parts of the biofilm surface, indicating the destruction of biofilms. In contrast, in the untreated control samples, the structured biofilms were flat and prominent. MTT assay indicated that CEN-ZnO NPs had less cytotoxicity on the MRC-5 lung fibroblast cells compared with the cancerous treated A549 cells. As the concentration of the CEN-ZnO NPs increased, the amount of ROS produced in the tested bacterial strains also increased. Analyzing the data obtained from flow cytometry showed that the higher concentrations of CEN-ZnO NPs lead to a reduction in the viability of P. aeruginosa PAO1, E. coli and S. aureus. The biosynthesized ZnO nanoparticles using Nostoc cell extracts exhibited different attributes, inspiring enough to be considered for further investigation.
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The development and spread of bacterial resistance to antimicrobial drugs necessitates the need to search for novel and effective antimicrobial agents. In the last few decades, innovative nanomaterials are attracting increasing attention and, among them, dendrimers have shown wide application in the various fields. In the current study, the two generations of an anionic linear- spherical nanodendrimer G1 and G2 were synthetized and compound G2 of nanodendrimer conjugated with erythromycin. The structures of the nanodendrimers were characterized by FTIR spectroscopy, zetasizer, and scanning electron microscopy (SEM). The antibacterial activity of the erythromycin-conjugated nanodendrimer and erythromycin alone were evaluated by the microdilution method against Staphylococcus aureus, S. epidermidis, S. saprophyticus, and Pseudomonas aeruginosa. The size of first and second generation of nanodendrimer, and the erythromycin-conjugated nanodendrimer was 75, 95, and 65.6â¯nm, respectively. The drug loading percentage of the nanodendrimer conjugates was obtained to be in 35.2%. In our study, the erythromycin-conjugated nanodendrimer showed significantly more bacteriostatic and bactericidal activities against all four studied bacteria than erythromycin alone. Our study's results highlight that the erythromycin-conjugated nanodendrimer is a highly effective agent against Gram positive and negative bacteria. The antibacterial properties of erythromycin combined with the targeting potential of the nanodendrimer can lead to sustained intracellular delivery of therapeutic agent.
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Antibacterianos/farmacología , Dendrímeros/química , Portadores de Fármacos , Eritromicina/farmacología , Nanopartículas , Antibacterianos/química , Química Farmacéutica/métodos , Composición de Medicamentos , Eritromicina/química , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Nanotecnología , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/crecimiento & desarrollo , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/crecimiento & desarrollo , Staphylococcus epidermidis/efectos de los fármacos , Staphylococcus epidermidis/crecimiento & desarrollo , Staphylococcus saprophyticus/efectos de los fármacos , Staphylococcus saprophyticus/crecimiento & desarrollo , Relación Estructura-ActividadRESUMEN
Influenza A virus is known worldwide as a threat associated with human and livestock diseases. Hence, identification of physiological and molecular aspects of influenza A could contribute to better design of therapeutic approaches for reducing adverse effects associated with disease caused by this virus. miRNAs are epigenetic regulators playing important roles in many pathological processes that help in progression of influenza A. Besides miRNAs, exosomes have ememrged as other effective players in influenza A pathogenesis. Exosomes exert their effects via targeting their cargos (e.g., DNAs, mRNA, miRNAs and proteins) to recipient cells. Here, we summarized various roles of miRNAs and exosomes in influenza A pathogenesis. Moreover, we highlighted therapeutic applications of miRNAs and exosomes in influenza.