Isolation and identification of nontuberculous mycobacteria from raw milk and traditional cheese based on the 16S rRNA and hsp65 genes, Tehran, Iran.

As an important source of human food, milk can be a carrier of human pathogenic bacteria, including tuberculous and nontuberculous mycobacteria (NTM), in its raw and unpasteurized state. In this research, 175 raw milk samples and 175 traditional cheese samples were collected from traditional dairy stores in 22 regions of Tehran in a 9- month period from August 2019 to May 2020. Samples were prepared and transferred to a specialized laboratory, where they were inoculated in Lowenstein-Jensen (LJ) medium containing glycerol or sodium pyruvate, as well as Herrold's egg-yolk with and without Mycobactin J. to determine the sample's identity of samples. The recommended 16S rRNA (1436 bp) and hsp65 (644 bp) gene fragments from the positive isolates identified in Ziehl-Neelsen (Z-N) staining were amplified and sequenced using PCR and compared with the sequences of the gene fragments of reference strains available in the global GenBank database. No mycobacterial species were isolated from traditional cheese samples in microbial culture. In case of raw milk samples, a total of four bacteria were collected, all of which were found in the genetic differential testing to be NTM, including n = 1 Mycobacterium heraklionense, n = 2 Mycolicibacterium fortuitum, and n = 1 Mycobacterium thermoresistibile. The analysis of the results obtained by isolate sequencing using the 16S rRNA gene showed higher discriminatory power and percentage similarities in the identification of the isolates than the hsp65 gene.

Expression of selected long non-coding RNAs in gastric cancer cells treated with coumarin: Possible mechanisms for anti-cancer activity.

Long non-coding RNAs (lncRNAs) can be utilized as prognostic indicators of gastric cancer since they can affect several cancer-related processes. Coumarin is a natural product with some useful anti-cancer properties. Here, we measured the expression of selected lncRNAs (RuPAR, SNHG6, CASC11, and their targets, miR-340-5p, p21, E-cadherin, and CDK1) in AGS gastric cancer cells treated with coumarin. MTT test has been utilized for assessing the AGS cells' cell viability after exposure to coumarin. The expression of the lncRNAs (RuPAR, SNHG6, and CASC11) and miR-340-5p was evaluated via qRT-PCR. Western blot analysis has been utilized to determine changes in p21, E-cadherin, and CDK1 expression. Coumarin decreased AGS viability in a dose-dependent manner. The coumarin treated cells had lower levels of the mRNAs known to be targets of lncRNAs SNHG6 and CASC11 compared to control. Additionally, the coumarin group had increased levels of lncRNA RuPAR expression when compared with the control group. Some lncRNA targets, including p21, E-cadherin, and CDK1, showed lower expression in the coumarin group compared to the control by Western blotting. Coumarin could be a promising pharmacological candidate to be included in gastric cancer treatment regimens because it modulates lncRNAs and their targets.

Characterization of green synthesized selenium nanoparticles (SeNPs) in two different indigenous halophilic bacteria.

BACKGROUND: In the biological method, using nonpathogenic and extremophile bacteria systems are not only safe and highly efficient but also a trump card for synthesizing nanoparticles. Halomonas elongata QW6 IBRC-M 10,214 (He10214) and Salinicoccus iranensis IBRC-M 10,198 (Si10198), indigenous halophilic bacteria, can be used for synthesizing selenium nanoparticles (SeNPs). METHODS: SeNP biosynthesis was optimized in two halophilic bacteria and characterized by UV-Vis, Fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), field emission scanning electron microscopy (FESEM), X-ray powder diffraction (XRD), zeta potential, and energy dispersive X-ray (EDX). RESULTS: Optimized conditions for synthesizing SeNPs was at 300 °C at 150 rpm for 72 h and 6 mM or 8 mM concentration of Na2SeO3. UV-Vis indicated a sharp absorption peak at 294 nm. Spherical-shaped nanoparticles by a diameter of 30-100 nm were observed in FESEM and TEM microscopy images. The produced SeNPs were identified by a peak in FTIR spectra. In XRD analysis, the highest peak diffraction had a relationship with SeNPs. The zeta potential analysis showed SeNP production, and elemental selenium was confirmed by EDX. CONCLUSIONS: Halophilic bacteria, owing to easy manipulation to create optimization conditions and high resistance, could serve as appropriate organisms for the bioproduction of nanoparticles. The biological method, due to effectiveness, flexibility, biocompatibility, and low cost, could be used for the synthesis of reproducible and stable nanoparticles.

More insights about genomic population structure of Mycobacterium avium subspecies paratuberculosis (Map) from multiple hosts in west and central provinces of Iran using a boosted genotyping approach.

To investigate the population genetic of Mycobacterium avium subsp. paratuberculosis (Map) in Iran, Mycobacterial Interspersed Repetitive Units (MIRUs) and Multi Locus Short Sequence Repeat (MLSSR) system were employed. Numerous genotypes by MIRU (N = 11) and MLSSR (N = 9) methods bearing discriminatory indices of 0.90 and 0.79 respectively, were obtained. Browsing the INRA-Nouzilly list (http://mac-inmv.tours.inra.fr/) detected 3 of the found patterns as new types. Some loci either MIRU-VNTR or SSR proved more polymorphic and therefore are recommended to be applied in priority for strain typing in the Iranian environment. While identical MIRU-VNTR or MLSSR patterns were detected among different conspecifics and geographical locations, dissimilar types were also observed at the same farms an indication of coexistence of Map strains within one herd. We suggest extension of the genotyping work described here to include more endogenous isolates in order to better analysis of transmission and virulence in epidemiology and control of paratuberculosis.

Interleukin-24-mediated antitumor effects against human glioblastoma via upregulation of P38 MAPK and endogenous TRAIL-induced apoptosis and LC3-II activation-dependent autophagy.

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.

Biosynthesis of Zinc Oxide Nanoparticles by Lactobacillus spp. and Investigation of their Antimicrobial Effect.

BACKGROUND: Nanoparticle biology is preferable to other common methods due to its economic efficiency and compatibility with the environment. On the other hand, the prevalence of drug-resistant bacterial strains is expanding and it is necessary to use alternative antibiotic compounds to deal with them. The aim of the present study was the biosynthesis of zinc oxide nanoparticles(ZnO NPs) by Lactobacillus spp. and their antimicrobial effect. METHODS: In this study, after the biosynthesis of ZnO NPs by Lactobacillus spp, Characterization of Nanoparticulation Was performed by UV-Vis, XRD, and Scanning Electron Microscopy (SEM). Additionally, Lactobacillus spp. - ZnO NPs were assessed for their antimicrobial properties. RESULTS: UV-visible spectroscopy confirmed the Lactobacillus spp. - ZnO NPs absorbed UV in the region of 300-400 nm. XRD analysis showed the presence of zinc metal in nanoparticles. SEM revealed that Lactobacillus plantarum - ZnO NPs were smaller than the others. Staphylococcus aureus showed the largest non-growth halo diameter against ZnO NPs synthesized by L. plantarum ATCC 8014 (3.7 mm). E. coli had the largest growth halo diameter against ZnO NPs synthesized by L. casei (3 mm) and L. plantarum (2.9 mm). The MIC values of ZnO NPs synthesized by L. plantarum ATCC 8014, L.casei ATCC 39392, L. fermenyum ATCC 9338, L. acidophilus ATCC 4356 were 2,8,8 and 4 µg/mL for Staphylococcus aureus. The MIC values of ZnO NPs synthesized by L. plantarum ATCC 8014, L. casei ATCC 39392, L. fermenyum ATCC 9338, L. acidophilus ATCC 4356 were 2, 4, 4, and 4 µg/ml for E. coli. The lowest MICs were 2 µg/ml for E. coli and S. aureus related to ZnO NPs synthesized by L. plantarum ATCC 8014. MIC and MBC values were equivalent to each other. CONCLUSION: The results of this research show that ZnO NPs synthesized by L. plantarum ATCC 8014 have more antimicrobial effects than other ZnO NPs used. Therefore, the ZnO NPs made with Lactobacillus plantarum ATCC 8014 have the potential to kill bacteria and can be considered a candidate for antibiotic replacement.

Antibacterial Effects of Nanocomposites on Efflux Pump Expression and Biofilm Production in Pseudomonas aeruginosa: A Systematic Review.

BACKGROUND: Pseudomonas aeruginosa is an opportunistic gram-negative pathogen with multiple mechanisms of resistance to antibiotics. This systematic review aimed to study the antibacterial effects of nanocomposites on efflux pump expression and biofilm production in P. aeruginosa. METHODS: The search was conducted from January 1, 2000, to May 30, 2022, using terms such as (P. aeruginosa) AND (biofilm) AND (antibiofilm activity) AND (anti-Efflux Pump Expression activity) AND (nanoparticles) AND (Efflux Pump Expression) AND (Solid Lipid NPS) AND (Nano Lipid Carriers). Many databases are included in the collection, including ScienceDirect, PubMed, Scopus, Ovid, and Cochrane. RESULTS: A list of selected articles was retrieved by using the relevant keywords. A total of 323 published papers were selected and imported into the Endnote library (version X9). Following the removal of duplicates, 240 were selected for further processing. Based on the titles and abstracts of the articles, 54 irrelevant studies were excluded. Among the remaining 186 articles, 54 were included in the analysis because their full texts were accessible. Ultimately, 74 studies were selected based on inclusion/exclusion criteria. CONCLUSION: Recent studies regarding the impact of NPs on drug resistance in P. aeruginosa found that various nanostructures were developed with different antimicrobial properties. The results of our study suggest that NPs may be a feasible alternative for combating microbial resistance in P. aeruginosa by blocking flux pumps and inhibiting biofilm formation.

Carotenoid pigment of Halophilic archaeon Haloarcula sp. A15 induces apoptosis of breast cancer cells.

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.

The Prevalence of bla VIM, bla KPC, bla NDM, bla IMP, bla SHV, bla TEM, bla CTX-M, and class I and II integrons Genes in Aeromonas hydrophila Isolated from Clinical Specimens of Qom, Iran.

BACKGROUND: Aeromonas hydrophila is an opportunistic gram-negative bacillus that causes diseases such as gastroenteritis, muscle infections, soft tissue, sepsis, and skin diseases in humans. Today, the prevalence of antibiotic resistance in bacteria has led to treatment failure and prolonged treatment. Therefore, the aim of this study was to evaluate the level of antibiotic resistance in isolates carrying bla VIM, bla KPC, bla NDM, bla IMP, bla SHV, bla TEM, bla CTX-M and class I and II integrons in Aeromonas hydrophila. METHODS: In this cross-sectional study, Aeromonas hydrophila were collected from different clinical specimens in Hazrat Masoumeh Hospital, Qom Province, Iran, from 2018 to 2020. The collected isolates were identified by standard biochemical tests. Then, using specific primers bla VIM, bla KPC, bla NDM, bla IMP, bla SHV, bla TEM, bla CTX-M genes, and class I and II integrons were evaluated by PCR method. Then, data were analyzed using SPSS software and chi-squared tests, and the significance level was determined as p ≤ 0.05. RESULTS: During the sample collection period, 100 Aeromonas hydrophila were collected. Based on the results of the antibiotic resistance pattern, the highest and lowest rate of antibiotic resistance to ampicillin (92%) and azithromycin (4%) were determined for both. Among the 100 isolates, 60 isolates produced broad-spectrum beta-lactamase (ESBL) and 50 isolates produced carbapenemase. Among the studied beta-lactamase genes, the highest and lowest frequencies were related to bla CTX-M (58%) and bla TEM (1%), respectively. The frequency of class I and II integron genes was 27% and 15%, respectively. CONCLUSIONS: The results of the study of antibiotic resistance, beta-lactamase, and carbapenemase genes showed high resistance in Aeromonas hydrophila, which raises concerns with regard to controlling infection in medical centers. Also, the study of antibiotic resistance in the presence of beta-lactamase genes showed that there was only a significant relationship between the presence of bla CTX-M gene and resistance to imipenem (p = 0.037).

Mixed pulmonary infection with four isolates of nontuberculous mycobacteria: a case report of mycobacterium bacteremicum infection.

BACKGROUND: A mixed pulmonary infection of Mycobacterium bacteremicum and three different isolates of nontuberculous mycobacteria (NTM) is an unusual clinical manifestation and have not yet been indicated. In this case report, we reported four isolates of NTM using phenotypic and genotypic test of pulmonary sample in Tehran, Iran. CASE PRESENTATION: We report a case of severe pulmonary disease in a 19-year-old male patient with productive cough, shortness of breath, and low-grade fever for several weeks. The C-reactive protein (CRP) level (80.2 mg/L) and erythrocyte sedimentation rate (ESR) (95 mm/h) were high. The computed tomographic scan indicated bronchiectasis, nodular opacities, consolidation, and cavitary lesions on both sides. The result of purified protein derivative (PPD) test was equal to 15 mm. The sequences of hsp65, rpoB, and 16S rDNA genes indicated more than 99% homology to four isolates of nontuberculous mycobacteria (NTM), including Mycobacterium fortuitum, M. chelonae, M. mucogenicum, and M. bacteremicum. We found that all four strains were susceptible to amikacin, cefoxitin, ciprofloxacin, clarithromycin, imipenem, and linezolid. The patient was treated with ciprofloxacin, clarithromycin, and amikacin, along with Montelukast, for five months. CONCLUSION: We report a case of severe pulmonary infection by four isolates of NTM. After treatment, the patient reported complete resolution of the signs and a weight gain of 5 kg; also, the CRP and ESR were normal. Nine months after the infection diagnosis, a new CT scan revealed further improvements.

Interrelation among exercise training, cardiac hypertrophy, and tissue kallikrein-kinin system in athlete and non-athlete women.

Introduction: The tissue kallikrein-kinin system is an endogenous homeostatic pathway, which its stimulation is associated with cardioprotection. The present study aimed to determine the effect of exercise training on plasma tissue kallikrein (TK) and bradykinin (BK) and their association with cardiac hypertrophy. Methods: 22 non-athlete and 22 athlete women were exposed to acute (Bruce test) and chronic (12-week swimming training) exercises. 2D echocardiography was used to evaluate morphological and functional features of the heart. Plasma concentrations of TK and BK were quantified by ELISA. Results: Athletes had significantly higher values of left ventricle end-diastolic diameter index (LVEDDI) and left ventricle mass index (LVMI) than non-athletes. Exercise intervention affected echocardiographic features in neither of the study groups. Chronic exercise training notably increased plasma levels of TK and BK, which increase was more pronounced in the athletes. Plasma TK negatively correlated with LVEDDI (r=-0.64, P=0.036 and r=-0.58, P=0.027) and LVMI (r=-0.51, P=0.032 and r=-0.63, P=0.028) in the non-athlete and athlete groups. In opposition, there was a positive correlation between plasma TK and left ventricle ejection fraction in non-athletes (r=0.39, P=0.049) and athletes (r=0.53, P=0.019). Conclusion: The upregulation of the tissue kallikrein-kinin system may be a protective mechanism against excessive cardiac hypertrophy induced by chronic exercise training.

Analysis of the A549 cell line affected by anticancer bioactive compounds of Actinomycetes isolated from saline soils.

Actinomycetes are filamentous bacteria and the residents of the soil, prone to produce bioactive metabolites. This research aimed to isolate, classify, and investigate the anticancer properties of Actinomycetes secondary metabolites from various saline soils of Qom province. Actinomycetes isolates were molecularly recognized by 16SrRNA gene sequencing after the PCR procedure. The A549 cell line was then exposed to bacterial metabolites to find their cytotoxicity by MTT assay and their capacity to cause apoptosis by Flow cytometry. The expression levels of the bax and bcl-2 genes were determined using Real-time PCR. Bacterial metabolites were distinct by HPLC and GC-MS assays. Sequencing identified three novel Actinomycetes strains, Streptomyces griseoflavus, Streptomyces calvus, and Kitasatospora phosalacineus. The IC50 doses of bacterial metabolites were discovered equal to 1337, 2619, and 4874 µg/ml, respectively. Flow cytometric assay revealed that their secondary metabolites were capable of inducing apoptosis in A549 cells by 25%, 14.5%, and 7.58%, respectively. Real-time PCR findings displayed that the bax gene expression in A549 cells treated with S. griseoflavus and S. calvus, comparatively increased (P < 0.0008, P < 0.00056). The expression of the bcl-2 gene was significantly reduced in cells treated with S. griseoflavus and K. phosalacineus (P < 0.0006, P < 0.0004). The findings of this analysis showed the presence of new isolates in a soil sample from Qom province which can produce new anticancer agents and can be considered appropriate candidates for further research to employ as anticancer drugs.

Enhanced synergistic antitumor effect of a DNA vaccine with anticancer cytokine, MDA-7/IL-24, and immune checkpoint blockade.

BACKGROUND: MDA-7/IL-24 cytokine has shown potent antitumor properties in various types of cancer without exerting any significant toxicity on healthy cells. It has also been proved to encompass pro-immune Th1 cytokine-like behavior. Several E7 DNA vaccines have developed against human papillomavirus (HPV)-related cervical cancer. However, the restricted immunogenicity has limited their clinical applications individually. To address this deficiency, we investigated whether combining the E7 DNA vaccine with MDA-7/IL-24 as an adjuvant would elicit efficient antitumor responses in tumor-bearing mouse models. Next, we evaluated how suppression of immunosuppressive IL-10 cytokine would enhance the outcome of our candidate adjuvant vaccine. METHODS: For this purpose, tumor-bearing mice received either E7 DNA vaccine, MDA-7/IL-24 cytokine or combination of E7 vaccine with MDA-7/IL-24 adjuvant one week after tumor challenge and boosted two times with one-week interval. IL-10 blockade was performed by injection of anti-IL-10 mAb before each immunization. One week after the last immunization, mice were sacrificed and the treatment efficacy was evaluated through immunological and immunohistochemical analysis. Moreover, the condition of tumors was monitored every two days for six weeks intervals from week 2 on, and the tumor volume was measured and compared within different groups. RESULTS: A highly significant synergistic relationship was observed between the E7 DNA vaccine and the MDA-7/IL-24 cytokine against HPV-16+ cervical cancer models. An increase in proliferation of lymphocytes, cytotoxicity of CD8+ T cells, the level of Th1 cytokines (IFN-γ, TNF-α) and IL-4, the level of apoptotic markers (TRAIL and caspase-9), and a decrease in the level of immunosuppressive IL-10 cytokine, together with the control of tumor growth and the induction of tumor regression, all prove the efficacy of adjuvant E7&IL-24 vaccine when compared to their individual administration. Surprisingly, vaccination with the DNA E7&IL-24 significantly reduced the population of Regulatory T cells (Treg) in the spleen of immunized mice compared to sole administration and control groups. Moreover, IL-10 blockade enhanced the effect of the co-administration by eliciting higher levels of IFN-γ and caspase-9, reducing Il-10 secretion and provoking the regression of tumor size. CONCLUSION: The synergy between the E7 DNA vaccine and MDA-7/IL-24 suggests that DNA vaccines' low immunogenicity can be effectively addressed by coupling them with an immunoregulatory agent. Moreover, IL-10 blockade can be considered a complementary treatment to improve the outcome of conventional or novel cancer therapies.

Isolating and sequencing vB_Kpn_3, a lytic bacteriophage against multidrug-resistant Klebsiella pneumoniae.

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.

Novel insertion mutation in the PLA2G6 gene in an Iranian family with infantile neuroaxonal dystrophy.

BACKGROUND: Infantile neuroaxonal dystrophy is an autosomal recessive neurological disorder. Individuals with infantile neuroaxonal dystrophy experience progressive loss of vision, mental skills and muscular control, and other variable clinical signs. Pathogenic variants in the PLA2G6 gene, encoding phospholipase A2, are recognized to be the fundamental reason for infantile neuroaxonal dystrophy. This study aimed to detect pathogenic variant in a consanguine Iranian family with infantile neuroaxonal dystrophy. METHODS: The mutation screening was done by whole exome sequencing followed by direct Sanger sequencing. RESULTS: We identified a homozygous insertion mutation, NM_003560: c.1548_1549insCG (p.G517Rfs*29) in exon 10 of PLA2G6 in the patient. The parents were heterozygous for variant. CONCLUSIONS: Because of the clinical heterogeneity and rarity of infantile neuroaxonal dystrophy, whole exome sequencing is critical to confirm the diagnosis and is an excellent tool for INAD management.

Desertifilum sp. EAZ03 cell extract as a novel natural source for the biosynthesis of zinc oxide nanoparticles and antibacterial, anticancer and antibiofilm characteristics of synthesized zinc oxide nanoparticles.

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.

Investigation of diverse biosynthetic secondary metabolites gene clusters using genome mining of indigenous Streptomyces strains isolated from saline soils in Iran.

Background and Objectives: Bioactive secondary metabolites are the products of microbial communities adapting to environmental challenges, which have yet remained anonymous. As a result of demands in the pharmaceutical, agricultural, and food industries, microbial metabolites should be investigated. The most substantial sources of secondary metabolites are Streptomyces strains and are potential candidates for bioactive compound production. So, we used genome mining and bioinformatics to predict the isolates secondary metabolites, biosynthesis, and potential pharmaceuticals. Materials and Methods: This is a bioinformatics part of our previous experimental research. Here, we aimed to inspect the underlying secondary metabolite properties of 20 phylogenetically diverse Streptomyces species of saline soil by a rationalized computational workflow by several software tools. We examined the Metabolites' cytotoxicity and antibacterial effects using the MTT assay and plate count technique, respectively. Results: Among Streptomyces species, three were selected for genome mining and predicted novel secondary metabolites and potential drug abilities. All 11 metabolites were cytotoxic to A549, but ectoine (p≤0.5) and geosmin (p≤0.001) significantly operated as an anti-cancer drug. Metabolites of oxytetracycline and phosphinothricin (p≤0.001), 4Z-annimycin and geosmin (p≤0.01), and ectoine (p≤0.5) revealed significant antibacterial activity. Conclusion: Of all the 11 compounds investigated, annimycin, geosmin, phosphinothricin, and ectoine had antimicrobial properties, but geosmin also showed very significant anti-cancer properties.

Exosomal long non-coding RNAs: novel molecules in gastrointestinal cancers' progression and diagnosis.

Gastrointestinal (GI) cancers arise in the GI tract and accessory organs, including the mouth, esophagus, stomach, liver, biliary tract, pancreas, small intestine, large intestine, and rectum. GI cancers are a major cause of cancer-related morbidity and mortality worldwide. Exosomes act as mediators of cell-to-cell communication, with pleiotropic activity in the regulation of homeostasis, and can be markers for diseases. Non-coding RNAs (ncRNAs), such as long non-coding RNAs (lncRNAs), can be transported by exosomes derived from tumor cells or non-tumor cells. They can be taken by recipient cells to alter their function or remodel the tumor microenvironment. Moreover, due to their uniquely low immunogenicity and excellent stability, exosomes can be used as natural carriers for therapeutic ncRNAs in vivo. Exosomal lncRNAs have a crucial role in regulating several cancer processes, including angiogenesis, proliferation, drug resistance, metastasis, and immunomodulation. Exosomal lncRNA levels frequently alter according to the onset and progression of cancer. Exosomal lncRNAs can therefore be employed as biomarkers for the diagnosis and prognosis of cancer. Exosomal lncRNAs can also monitor the patient's response to chemotherapy while also serving as potential targets for cancer treatment. Here, we discuss the role of exosomal lncRNAs in the biology and possible future treatment of GI cancer.

vB-Ea-5: a lytic bacteriophage against multi-drug-resistant Enterobacter aerogenes.

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.

Protective cellular and mucosal immune responses following nasal administration of a whole gamma-irradiated influenza A (subtype H1N1) vaccine adjuvanted with interleukin-28B in a mouse model.

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.