Antibacterial activity of Lake Urmia derived-Halomonas.

Isolation of new microbial species from extreme environments is one of the most efficient approaches for the development of novel bioactive metabolites. The aim of the present study was to explore the pharmaceutical bacterial resources from the water and sediments of hypersaline Lake Urmia. Using different culture conditions and media led to the isolation of 20 bacterial strains. Halophilic bacteria were screened for the production of antibacterial agent against multi-drug resistant strains of Escherichia coli through agar well diffusion assay. Halophilic bacteria DNA extraction was done by boiling method. The results showed that two Halomonas strains, LUH16 and LUH20 identified by analysis of 16S rRNA gene sequences were the potent producers of antimicrobial metabolites against various strains of E. coli. Furthermore, gas chromatography-mass spectrometry (GC-MS) analysis revealed the presence of eight secondary metabolites with the relevant antimicrobial properties. Our findings led us to focus on Halomonas strains as potent producers of antimicrobial compound that might be an alternative against antibiotic-resistant pathogens such as pathogenic Escherichia coli.

An Ultrasensitive miRNA-Based Genosensor for Detection of MicroRNA 21 in Gastric Cancer Cells Based on Functional Signal Amplifier and Synthesized Perovskite-Graphene Oxide and AuNPs.

In the present research work, the state-of-art label-free electrochemical genosensing platform was developed based on the hybridization process in the presence of [Fe(CN)6]3-/4- as an efficient redox probe for sensitive recognition of the miRNA-21 in human gastric cell lines samples. To attain this aim, perovskite nanosheets were initially synthesized. Afterward, the obtained compound was combined with the graphene oxide resulting in an effective electrochemical modifier, which was dropped on the surface of the Au electrode. Then, AuNPs (Gold Nano Particles) have been electrochemically-immobilized on perovskite-graphene oxide/Au-modified electrode surface through the chronoamperometry (CA) technique. Finally, a self-assembling monolayer reaction of ss-capture RNA ensued by the thiol group at the end of the probe with AuNPs on the modified electrode surface. miRNA-21 has been cast on the Au electrode surface to apply the hybridization process. To find out the effectiveness of the synthesized modifier agent, the electrochemical behavior of the modified electrode has been analyzed through DPV (differential pulse voltammetry) and CV (cyclic voltammetry) techniques. The prepared biomarker-detection bioassay offers high sensitivity and specificity, good performance, and appropriate precision and accuracy for the highly-sensitive determination of miRNA-21. Different characterization methods have been used, such as XRD, Raman, EDS, and FE-SEM, for morphological characterization and investigation of particle size. Based on optimal conditions, the limit of detection and quantification have been acquired at 2.94 fM and 8.75 fM, respectively. Furthermore, it was possible to achieve a wide linear range which is between 10-14 and 10-7 for miRNA-21. Moreover, the selectivity of the proposed biosensing assay was investigated through its potential in the detection of one, two, and three-base mismatched sequences. Moreover, it was possible to investigate the repeatability and reproducibility of the related bio-assay. To evaluate the hybridization process, it is important that the planned biomarker detection bio-assay could be directly re-used and re-generated.

Developing erythromycin resistance gene by heavy metals, Pb, Zn, and Co, in aquatic ecosystems.

Industrial development is the main cause of environmental pollution with various substances such as antibiotics and heavy metals. Many heavy metals with antimicrobial properties could contribute to antibiotic resistance and the emergence of antibiotic resistance genes due to the co-selection phenomenon. The aim of this study was to investigate the concurrent presence and correlation between several heavy metals and the erythromycin resistance genes in six aquatic ecosystems of Iran. Distribution and assessment of 11 erythromycin resistance genes were investigated using specific primers and online enrichment and triple-quadrupole LC-MS/MS. The concentration of heavy metals was measured using inductively coupled plasma atomic emission spectroscopy by Thermo electron corporation. Principal component analysis was performed to globally compare and to determine the similarities and differences among different aquatic ecosystems in different parts of the world in terms of the concentration of zinc and lead in their water. The results of the simple logistic regression analysis for the correlation between erythromycin resistance genes and heavy metals concentrations revealed the most significant correlation between erythromycin resistance genes and Pb concentration, followed by Co and Zn concentrations.

Isolation of a lytic bacteriophage against extensively drug-resistant Acinetobacter baumannii infections and its dramatic effect in rat model of burn infection.

OBJECTIVES: Acinetobacter Baumannii is an opportunistic nosocomial pathogen belonging to the Moraxellaceae family. The emergence of multidrug resistant strains of this pathogen caused many problems for hospitals and patients. The aim of the current study was to isolate, identify, and morphologically, physiologically, and in vivo analyze a new lytic bacteriophage targeting extensively drug-resistant (XDR) A. baumannii. MATERIALS AND METHODS: Different wastewater samples were tested for isolation of lytic bacteriophage against 19 A. baumannii isolates obtained from patients hospitalized in a hospital in Arak, Iran, from January 2019 to March 2019. The phenotypic and genotypic characteristics of A. baumannii strains (resistance genes including: adeA, adeB, adeC, adeR, adeS, ISAba1, blaOXA-23, blaOXA-24) were analyzed. The isolated phage characteristics including adsorption time, pH and thermal stability, host range, one-step growth rate, electron microscopy examination, and therapeutic efficacy of the phage were also investigated. Therapeutic efficacy of the phage was evaluated in a rat model with burn infection of XDR A. baumannii. The lesion image was taken on different days after burning and infection induction and was compared with phage untreated lesions. RESULTS: The results showed unique characteristics of the isolated phage (vB-AbauM-Arak1) including high specificity for Acinetobacter baumannii, stability at a relatively wide range of temperatures and pH values, short adsorption time, short latent period, and large burst size. In relation to the therapeutic efficacy of the phage, the lesion area decreased in phage-treated groups over 14 days than in those untreated, significantly (p < 0.05). CONCLUSION: Our findings demonstrated that isolated lytic phage was able to eliminate burn infections caused by XDR A. baumannii in a rat model. So, it may be recommended as alternative options toward to developing a treatment for extensively drug resistant Acinetobacter infections.

Association of Herpes simplex virus I&II infections with rs187084 SNP of TLR9 and male infertility.

Since TLR9 recognises unmethylated CpG motifs in viral DNA, its polymorphisms may contribute to the susceptibility to Herpes simplex virus I&II infection. In the present study, to evaluate the role of rs187084 SNP (single nucleotide polymorphism) of TLR9 in Herpes simplex virus I&II infection and male infertility, 103 infertile and 27 fertile blood and semen samples were analysed. We assessed the micro and macro properties of semen specimens and the presence of HSV immunoglobulins. Tetra-primer ARMS PCR was used to detect SNP and to investigate the genotype distribution of TLR9-rs187084 SNPs, and the correlation between polymorphisms of TLR9 gene and male infertility. Moreover, the odds ratio (OR) and 95% confidence intervals were used to estimate the strength of the association. Based on our finding, a significant correlation was observed between HSV infection, agglutination and polymorphism (TT) under dominant (OR = 1.28, 95% CI = 0.94-1.75) and recessive (OR = 0.44, 95% CI = 0.21-0.94) models for the data, which was complied with Hardy-Weinberg equilibrium (HWE) (OR = 2.91, 95% CI = 1.02-8.30). The result showed a significant association between HSV IgM and agglutination in HSV infection (p < .001), and in addition, there were associations between alleles so that rs187084 SNP might be considered as a risk factor for the incidence of HSV infection.

Cytotoxic effect of sea anemone pore-forming toxin on K562 chronic myeloid leukemia cells.

Chronic myelogenous leukemia (CML) is one of prevalent cancer worldwide. In spite of various designed drugs, chemoresistance remains the main obstacle in cancer cure. Therefore, developing novel strategy for treatment of CML is an urgent need. Fragaceatoxin C (FraC) is novel protein toxin from a sea anemone called actinia fragacea with great impacts against cells by pore formation and disturbing cell membrane integrity. The aim of this study was evaluation of FraC toxin toxicity against K562. The bacteria cells harboring expression||||||| vector of FraC were induced by IPTG and purified by Ni2+-NTA sepharose affinity chromatography. Then, purified toxin activity was evaluated using RBC hemolytic test. Eventually, evaluation of FraC cytotoxicity and apoptosis were performed using MTT and flow cytometery assays, respectively. Our results revealed that FraC toxin decreased K562 cells viability in a dose- and time-dependent manner with a whole destroy of cancer cells at 35.00 µg mL-1 after 72 hr. Furthermore, flow cytometery analysis indicated that FraC toxin enhanced necrosis along with apoptosis in K562 cells in a dose dependent manner. We speculated that FraC toxin could be considered as a novel candidate for cancer cell researches and treatments provided that it should be turned into a specific agent by engineering and directing to cancer cell membrane.

Comparing the Bacteriostatic Effects of Different Metal Nanoparticles Against Proteus vulgaris.

For many years, researchers were looking for new antibacterial substances to deal with hospital infections and especially resistant infections. Nanoparticles attracted much attentions because of their very small size that increases the surface to capacity ratio and consequently increase chemical activity. In this study, the antibacterial effects of silver, copper oxide, nickel oxide, and titanium dioxide nanoparticles were studied on Proteus vulgaris, as a bacterium involved in the resistant hospital infections. The capability of nanoparticles to inhibit the growth of bacteria was assessed via 9 different methods including cylinder, disk, and well-diffusion, spot test, MBC, MIC, liquid inhibitory action test, diffusion, and assessing the effects of nanoparticles on a 24-h culture. Based on the results, copper oxide and silver nanoparticles had high antibacterial effects on P. vulgaris in both liquid and solid cultures, respectively. However, nickel oxide and titanium dioxide nanoparticles only had a weak effect on the inhibition of bacterial growth in the liquid culture. CuO and Ag NPs could release ions and consequently produce free radicals, disturb the equilibrium of electrons between electron donor groups and inactivate enzymes and DNA of the organisms. Moreover, they triggered holes in the bacterial membrane to disturb cellular ion equilibrium. So, they can be used to inhibit the growth of pathogens. Besides, further studies have shown that they could be used as a supplementary treatment and/or in combination with other drugs to cure infections caused by P. vulgaris.

Identification of Genomic Species of Acinetobacter Isolated from Burns of ICU Patients.

BACKGROUND: The worldwide emergence of multi-drug resistant (MDR) bacteria in recent years has caused many problems for hospitals and patients, especially intensive care unit patients. Among these clinically important MDR bacteria are Acinetobacter baumannii complex species (A. baumannii, Acinetobacter genomic species 3 and Acinetobacter genomic species 13TU) that cause a wide range of infections. METHODS: The sequencing and bioinformatics analysis of a part of the Zone 1 of rpoB gene was performed for species identification of Acinetobacter isolates obtained from ICU patients with infected burns hospitalized in a hospital in Isfahan, Iran, over a 9-month period. Antibiotic sensitivity of Acinetobacter isolates was investigated using the disk diffusion method and different classes of antibiotics including amikacin, cefotaxime, ceftriaxone, ciprofloxacin, imipenem and piperacillin. RESULTS: Acinetobacter spp. were isolated from 10 of 80 (12.5%) investigated patients. All of the 10 Acinetobacter isolates were identified as Acinetobacter baumannii and multi-drug resistant according to antibiotic susceptibility tests. CONCLUSION: Of the Acinetobacter baumannii complex members, only A. baumannii species was identified among the isolates obtained from patients with infected burns in an Isfahan hospital over a 9-month period.

HIV-1 Vpr Protein Enhances Proteasomal Degradation of MCM10 DNA Replication Factor through the Cul4-DDB1[VprBP] E3 Ubiquitin Ligase to Induce G2/M Cell Cycle Arrest.

Human immunodeficiency virus type 1 Vpr is an accessory protein that induces G2/M cell cycle arrest. It is well documented that interaction of Vpr with the Cul4-DDB1[VprBP] E3 ubiquitin ligase is essential for the induction of G2/M arrest. In this study, we show that HIV-1 Vpr indirectly binds MCM10, a eukaryotic DNA replication factor, in a Vpr-binding protein (VprBP) (VprBP)-dependent manner. Binding of Vpr to MCM10 enhanced ubiquitination and proteasomal degradation of MCM10. G2/M-defective mutants of Vpr were not able to deplete MCM10, and we show that Vpr-induced depletion of MCM10 is related to the ability of Vpr to induce G2/M arrest. Our study demonstrates that MCM10 is the natural substrate of the Cul4-DDB1[VprBP] E3 ubiquitin ligase whose degradation is regulated by VprBP, but Vpr enhances the proteasomal degradation of MCM10 by interacting with VprBP.

Insights into new bacteriophages of Lactococcus garvieae belonging to the family Podoviridae.

Lactococcus garvieae is an emerging pathogen responsible for lactococcosis, a serious disease in trout aquaculture. The identification of new bacteriophages against L. garvieae strains may be an effective way to fight this disease and to study the pathogen's biology. Three L. garvieae phages, termed WP-1, WWP-2 and SP-2, were isolated from different environments, and their morphological features, genome restriction profiles and structural protein patterns were studied. Random cloning of HindIII-cut fragments was performed, and the fragments were partially sequenced for each phage. Although slight differences were observed by transmission electron microscopy, all of the phages had hexagonal heads and short non-contractile tails and were classified as members of the family Podoviridae. Restriction digestion analysis of the nucleic acids of the different phages revealed that the HindIII and AseI digests produced similar DNA fragment patterns. Additionally, SDS-PAGE analysis indicated that the isolated phages have similar structural proteins. The sequence BLAST results did not show any significant similarity with other previously identified phages. To the best of our knowledge, this study provides the first molecular characterization of L. garvieae phages.

Detection of torque teno virus (TTV) in domestic village chickens in Iran.

Torque teno virus (TTV) is prevalent worldwide and has been extensively studied in human and some wild and domestic animals. As the studies on TTV in chickens was rare and there was no information about the infection of domestic village chickens with TTV and also structural resemblance of this virus to chicken anemia virus, the frequency of the infection in domestic village chickens in different villages in Isfahan (Iran) was investigated. Sera were collected from 50 chickens. Viral DNA was extracted and subjected to polymerase chain reaction (PCR) using the previously described T801 and T935 primers that were used for amplification of a highly conserved non-coding region (UTR) of the viral genome in a single round of PCR and Set B primers of conserved region in a nested PCR reaction. Using T801 and T835 primers TTV or viruses of TTV family were detected in 16 out of 50 sera tested (32%). Fourteen out of the same 50 sera (28%) were positive for TTV using Set B primers. Totally 20 sera were positive using both primers (40%). Ten sera were detected with both sets of primers, six sera with T801 and T935 primers and only four sera were positive using Set B primers for TTV. Different patterns of the detection of the virus with the two different sets of primers suggests the possibility of the presence of different genotypes of TTV in domestic village chickens and the possibility of the transmission of the virus from human to village chickens and vice versa. This necessitates further investigations.

Torque teno mini virus infection and multiple sclerosis.

Multiple sclerosis (MS) is a disease of young adults which is characterized by autoimmune demyelination of the central nervous system. Interaction of genetics and environmental factors are required to cause MS. Among the proposed environmental factors for MS, viral infections are thought to play a role in the pathogenesis of the disease. Torque teno mini virus (TTMV), which has recently been shown to infect humans, is a member of circoviridae, and has a circular DNA with 2860 nucleotides. Since there are a few data about the pathogenicity of this virus, this study sought to investigate the presence of TTMV in sera from MS patients and healthy individuals. We studied 149 serum samples from MS patients and 150 sera of healthy individuals. Serum DNA was extracted using phenol-chloroform and was subjected to nested polymerase chain reaction. TTMV-DNA was detected in 24 (16%) sera of the healthy blood donors and in 21 (14.1%) samples of the MS patients, where the difference did not reach significance (p > .05). The result of this study could not establish an association between TTMV infection and MS.