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Background: In the development of multidrug resistance, efflux pumps effectively pump drug compounds out of cells, which results in reduced membrane permeability to drug compounds. This study evaluated the effect of gold nanoparticles on the inhibition of norA and norB efflux pumps in ciprofloxacin-resistant Staphylococcus aureus isolated from burn patients in Qom province, Iran. Methods: In this cross-sectional study, S. aureus strains were isolated from burn patients in Qom hospital, Iran. After gold nanoparticles were synthesized using chemical reduction and characterized by spectrophotometry, transmission electron microscopy (TEM), and dynamic light scattering (DLS), ciprofloxacin resistance of S. aureus was screened by the disc diffusion method. The Minimum Inhibitory Concentration (MIC) of ciprofloxacin (CCCP), ciprofloxacin + gold nanoparticles (CCCP + gold nanoparticles), and ciprofloxacin + CCCP was determined. Moreover, norA and norB genes were evaluated by PCR using special primers. Real-time PCR was then performed for norA and norB genes. Results: Of 88 S. aureus strains tested, 50 (56.81%) were resistant to ciprofloxacin. From the 50 ciprofloxacin-resistant S. aureus strains, 12 isolates had active pumps. Real-time PCR of 12 ciprofloxacin- resistant S. aureus and S. aureus ATCC 25923 before and after exposure to ciprofloxacin, gold nanoparticles, and gold nanoparticles with sub-MIC ciprofloxacin revealed significant differences in expression of norA and norB genes before exposure to the treatments compared to after exposure (p Conclusion: Gold nanoparticles with ciprofloxacin could be used to prevent the expression of pump genes involved in resistance to fluoroquinolone compounds.

HNF4α, a member of the nuclear receptor superfamily, regulates the genes involved in lipid and glucose metabolism. The expression of the RARβ gene in the liver of HNF4α knock-out mice was higher versus wildtype controls, whereas oppositely, RARβ promoter activity was 50% reduced by the overexpression of HNF4α in HepG2 cells, and treatment with retinoic acid (RA), a major metabolite of vitamin A, increased RARβ promoter activity 15-fold. The human RARβ2 promoter contains two DR5 and one DR8 binding motifs, as RA response elements (RARE) proximal to the transcription start site. While DR5 RARE1 was previously reported to be responsive to RARs but not to other nuclear receptors, we show here that mutation in DR5 RARE2 suppresses the promoter response to HNF4α and RARα/RXRα. Mutational analysis of ligand-binding pocket amino acids shown to be critical for fatty acid (FA) binding indicated that RA may interfere with interactions of FA carboxylic acid headgroups with side chains of S190 and R235, and the aliphatic group with I355. These results could explain the partial suppression of HNF4α transcriptional activation toward gene promoters that lack RARE, including APOC3 and CYP2C9, while conversely, HNF4α may bind to RARE sequences in the promoter of the genes such as CYP26A1 and RARβ, activating these genes in the presence of RA. Thus, RA could act as either an antagonist towards HNF4α in genes lacking RAREs, or as an agonist for RARE-containing genes. Overall, RA may interfere with the function of HNF4α and deregulate HNF4α targets genes, including the genes important for lipid and glucose metabolism.

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.

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.

In this paper, we detail how damage mechanisms commonly found on marine terminal structures and process equipment can be automatically detected using a machine learning (ML) pipeline. Defects in its varied forms, such as corrosion, steel, wood and concrete damages must be closely monitored in offshore oil and gas platforms and also in marine terminals; the cost of not monitoring and detecting defects can range from unplanned shutdowns, plant damage, to human fatalities. On-site inspection of defects is labor intensive and require frequent travel to offshore and onshore facilities. This process has two shortcomings: firstly, it is costly involving man hours spent on the facility and secondly only a subset of the total defects is monitored during each inspection due to difficulty in obtaining full coverage; specially in this scenario a high-risk defect could be missed completely. We use marine terminals as an example of how defects can be detected using machine learning techniques, however the pipeline for generating the ML models could be extended to other types of defects and structures. Significantly, the output from our ML pipeline can be used as an input to the Risk Based Inspection (RBI) system assisting asset operators in their inspection and maintenance planning. Furthermore, digitized inspections and ML can provide around 95% coverage of the terminal while avoiding problems faced by a human inspector such as producing consistent results across inspections and sites. Our system captured terrestrial spherical imagery and range data (LIDAR), subsea imagery, sonar, and ultrasonic thickness (UT) gauge data using different sensors and equipment on a marine terminal. Equipment used in our data-capture included an unmanned aerial vehicle (UAV), a terrestrial scanner, a remotely operated vehicle for underwater imaging (ROV) and a remotely operated surface vehicle (ROSV). For the purposes of this work, we used the images captured by the terrestrial scanner. We built an automated defect detection pipeline using state of the art machine learning and artificial intelligence (AI) tools taking the cube face images as inputs. The defect detection pipeline was trained with examples of defects labeled by asset integrity engineers to ensure quality of results. We built the pipeline to detect damage mechanisms for steel and wooden structures, however our system could be scaled to incorporate other types of defects and structures. Our AI pipeline was able to achieve a high level of accuracy identifying wood damage with a mean-Average Precision (m-AP) of 0.80 and steel damage with a m-AP of 0.84, where a m-AP of 1.0 indicates perfect detection of damage.

Since recent research indicates that the addition of graphene increases the mechanical and biological properties of calcium phosphates simultaneously, graphene oxide (GO) was used in this research to enhance the properties of brushite cement. The main objective of this study is to investigate the mechanism of nucleation and growth of brushite crystals on GO sheets. Calcium nitrate tetrahydrate and diammonium hydrogenphosphate were used as calcium and phosphate precursors. The brushite chemical precipitation method on graphene sheets was used in this study and computer-assisted modeling was used to understand the process. The results of this study showed that the hybrid powders contained brushite and GO. Brushite crystals were grown after nucleation in proportion to the three main planes containing (020), (12 1 ¯ ), and (141). The final particles were plate shaped. The results of setting time tests showed that increasing the GO% decreases the setting time and this trend continues with an increasing amount of GO. The results of the mechanical evaluation showed that increasing the GO by up to 2% increased the mechanical properties and more than that decreased the mechanical properties.

Background: Bacillus clausii is being studied as a probiotic candidate. There is insufficient information on the antimicrobial and anticancer effects of B. clausii. Objectives: The present investigation was designed to evaluate the anti-bacterial, anti-adenoviral, and apoptosis-inducing activity of B. clausii cell-free supernatant (CFS). Methods: First, the supernatant of B. clausii was collected after culture for 24 h. Then, its anti-bacterial impact on several genera of bacteria was assessed through the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC). Adenovirus 5 (Ad5) was exposed to the CFS under four conditions, including pre-treatment: First infecting cells with CFS and then with the virus; pre-incubation: Incubation of the supernatant and virus for 1.5 hours and then adding to the cells; competition: Infection of cells with the simultaneous mixture of the supernatant and virus, and post-treatment: First infecting cells with the virus and then with CFS. The median tissue culture infectious dose (TCID50) technique determined the virus titer. Real-time PCR was performed to assess the E1A expression. After exposure to the CFS, real-time PCR was utilized to measure the expression of MicroRNA-145, BCL-2, and BAX in HeLa cancer cells. Results: Bacillus clausii supernatant showed an inhibitory effect on Methicillin-resistant Staphylococcus aureus (MRSA), Enterococcus faecalis, S. aureus, Pseudomonas aeruginosa, Escherichia coli, and Acinetobacter baumannii. The Ad5 titers were reduced by about 4.61, 4, 3.9, and 3.1 Log10 TCID50/mL in pre-treatment, pre-incubation, competition, and post-treatment tests (CFS dilution: 1/4), respectively. Similar results of the viral titration were seen when experimental and control E1A expression levels were compared. Also, B. clausii supernatant during 48 h exposure to HeLa cells increased the transcript of the BAX, BCL-2, and miR-145 genes to 9.1, 2.3, and 55 folds, respectively, compared to the untreated condition. Conclusions: Bacillus clausii can be a potent antimicrobial and anticancer agent. Further research is required to learn about the spectrum of anti-bacterial, antiviral, and anti-cancerous activities of B. clausii.

Background Coronavirus disease 2019 was spread worldwide, as a pandemic, from December 2019. Venous thromboembolism events can inflict patients with coronavirus disease 2019 during the hospitalization or convalescent period. Therefore, monitoring of these patients, in terms of venous thromboembolism events signs and symptoms, and timely management of antithrombotic agents are of great importance. Case report A 45-year-old Iranian man, who is the first author of this case report, was infected by severe acute respiratory syndrome coronavirus 2 and displayed the typical signs and symptoms of coronavirus disease 2019. Although reverse transcription polymerase chain reaction for coronavirus disease 2019, and specific immunoglobulin M and immunoglobulin G against severe acute respiratory syndrome coronavirus 2, were negative at first, chest computed tomography scan showed the characteristic pattern of lung involvement of a coronavirus disease 2019 infection including bilateral and multilobar ground-glass opacities. At that time, there were no signs or symptoms of deep-vein thrombosis or pulmonary thromboembolism, so these were not investigated. About 30 hours after hospital discharge, the patient presented back to the hospital with acute-onset chest pain. We instantly tested his blood for D-dimer, and sent him to take a Doppler sonography of his lower legs and a chest computed tomography angiography in search of pulmonary thromboembolism and deep-vein thrombosis. Although we could confirm pulmonary thromboembolism with computed tomography angiography in our patient, there were no signs or symptoms of venous thromboembolism in his lower legs, and color Doppler sonography of lower limbs was normal. So, the patient was treated with rivaroxaban as an antithrombotic agent. After some days, he was discharged in good condition. About 1 month later, he was referred to our hospital because of left lower limb edema. Although he was under antithrombotic therapy, color Doppler sonography of lower limbs revealed acute deep-vein thrombosis of the left leg. Hence, we decided to shift antithrombotic therapy from rivaroxaban to warfarin, as it is more potent than rivaroxaban in recurrent venous thromboembolism and when taking new oral anticoagulants. Unlike rivaroxaban, which needs no blood test to monitor its efficacy but has a warning for signs and symptoms of bleeding, warfarin therapy must be monitored carefully by regular blood tests for prothrombin time and international normalized ratio to maintain them in the therapeutic range. The patient was informed about the bleeding cautions, and required regular check of prothrombin time and international normalized ratio to maintain them in the proper and advised range of treatment (international normalized ratio therapeutic range 2–3). Conclusion In the case of unexpected recurrent venous thromboembolism in coronavirus disease 2019, especially when patients are taking rivaroxaban or other new oral anticoagulants, such drugs should be substituted by warfarin, with routine follow-up, to maintain the value of prothrombin time and international normalized ratio within the therapeutic range.

The halophilic microorganisms living in Extreme environments contain high concentrations of carotenoids with notable medical abilities. We evaluated the anti-cancer activity 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 eight strains capable of producing pigmented colonies were chosen from among those cultured strains. In the next step, from among eight strains using MTT assay, one of them capable of reducing cell viability of the breast cancer MCF-7 cell line was chosen as a selective strain. The principal carotenoid was characterized based on the 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 showed the strongest cytotoxic effect on breast cancer cell line (IC50 = 0.0625 mg/mL). 16S rRNA analysis indicated that strain A15 showed similarity with Haloarcula hispanica for about 99.5%. According to the results of the analysis, we can estimate that the principal carotenoid extracted form Haloarcula sp. strain A15 was similar to bacterioruberin. Both late and early apoptosis were increased significantly about 10% and 39%, respectively as a result of up-regulation of CASP3, CASP8, BAX genes expression in MCF-7 cell line. In contrast, the expression of genes MKI67, SOX2 were significantly down-regulated in treated MCF-7 cell line. Halophilic archaeon strain could be a good option to produce high added-value bacterioruberin due to its possible anti-cancer properties.

Regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs) are common immunosuppressive cells in the tumor microenvironment. These cells, through various mechanisms, inhibit antitumor immune responses and impede effective therapies. Therefore, designing an efficient protocol for inducing immune surveillance in tumors is highly recommended. Recently, nanoliposomes have provided broad-spectrum and state-of-the-art vehicles to deliver antigens or immune system compartments in immunotherapies. It has been shown that different lipids in the structure of liposomes and various liposomal formulations can affect immune responses in the tumor microenvironment. This study was aimed to evaluate the effects of four different liposomal formulations on MDSCs and Tregs in C26 tumor-bearing mice. To this end, after preparing liposomes, they were injected into tumor-inoculated mice and analyzed MDSC and Treg population and functions in spleen and tumor tissues. Results showed that DOTAP-containing liposomes reduced MDSC population and activity in the spleen, but not tumor, compared with other groups significantly (P

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.

Background and Objectives: Bioactive secondary metabolites are the products of microbial communities adapting to en- vironmental challenges, which haveyet remained anonymous. As a result of demands in the pharmaceutical, agricultural, and food industries, microbial metabolites should be investigated. Themost substantial sources of secondary metabolites are Streptomyces strains and are potential candidates for bioactive compound production. So, we usedgenome 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 rational- ized computational workflow by several software tools. Weexamined 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 11metabolites were cytotoxic to A549, but ectoine (p≤0.5) and geosmin (p≤0.001) signifi- cantly 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.

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.

Background Periprocedural myocardial injury is a predictor of cardiovascular morbidity and mortality after percutaneous coronary intervention. Methods The authors examined the effects of preprocedural lipid levels (low-density lipoprotein, high-density lipoprotein, and triglycerides) in 977 patients with coronary artery disease who underwent elective percutaneous coronary intervention. Results Elevated cardiac troponin I level (≥5× the upper limit of normal) was used to indicate periprocedural myocardial injury. Serum lipid samples were collected 12 hours preprocedurally. Cardiac troponin I was collected 1, 6, and 12 hours postprocedurally. Correlations between preprocedural lipid levels and postprocedural cardiac troponin I were studied. Low-density lipoprotein levels were less than 70 mg/dL in 70% of patients and greater than 100 mg/dL in only 7.4% of patients; 13% had triglyceride levels greater than or equal to 150 mg/dL, and 96% had high-density lipoprotein levels less than 40 mg/dL. Patients with elevated cardiac troponin I had significantly lower left ventricular ejection fraction than did those with cardiac troponin I levels less than 5× the upper limit of normal (P = .01). Double-and triple-vessel disease were more common in patients with elevated cardiac troponin I (P < .002). Multivariable logistic and linear regression analyses revealed no statistically significant associations between lipid levels and postprocedural cardiac troponin I elevation, possibly because such large proportions of included patients had low levels of low-density lipoprotein (70%) and a history of statin intake (86%). Conclusion The authors found no association between lipid profile and periprocedural myocardial injury.

With the increased usage of nitrate fertilizers, the removal of their stable ionic and water-soluble end products is a challenge for human health. Several physicochemical methods have been examined for nitrate removal of water, but biological treatments are mostly preferred due to a higher efficiency and lower cost. To remove nitrogen from water, we investigated the potential of nitrate-reducing halophilic and halotolerant bacteria. A total of 50 strains from different saline and hypersaline environments of Iran, including the Incheboron wetland, Aran-Bidgol salt-lake, and Urmia endorheic salt-lake, were screened for nitrate reductase production. Among investigated bacteria, 60% and 19% of strains obtained from Urmia lake, and Incheboron wetland produced nitrate reductases, respectively. The nitrate reductase coding genes narG, and napA were analyzed in all strains with confirmed nitrate-reducing capacity. The napA gene was successfully amplified from a gram-negative halophilic strain, and the narG gene was detected in ten halophilic strains. Among nitrate-reducing isolates with the narG gene expression, the Kocuria rosea strain R3A34 showed the highest nitrate reductase production level. This strain was selected to optimize for its denitrifying activity. Results showed that 32°C, pH 7.0, NaCl 8% (w/v), and mannitol (as a carbon source) provide the optimal environmental conditions for the efficient production of nitrate reductase by the Kocuria rosea strain R3A34. As these are compatible with wastewaters conditions, this bacterium can be a proper candidate for bioremediation of wastewaters from nitrate pollutants.

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.

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 IC

Background: One of the most common and treatable causes of stroke is carotid artery stenosis. Carotid artery stenting (CAS) is an option for treating the stenosis, with such notable clinical outcomes as low rates of in-hospital death, stroke, and intracerebral hemorrhage.Methods: All patients who underwent carotid angioplasty between December 2010 and January 2019 in Rajaie Cardiovascular Medical and Research Center were enrolled. The incidence rates of major adverse cardiac and cerebral events (MACCE), defined as a composite of stroke, myocardial infarction, bleeding, and all-cause mortality, both during hospitalization and at long-term follow-up were determined. Results: A total of 380 patients, 264 (69.5%) symptomatic and 116 (30.5%) asymptomatic, were included. The mean age of study population was 68.6 ± 10.6 years. During post-CAS in-hospital course, stroke occurred in 2 (0.5%), intracranial hemorrhage in 4 (1.1%), death in 2 (0.5%) and MACCE in 8 (2.1%) in symptomatic, and stroke in 1 (0.3%), intracranial hemorrhage in 1 (0.3%), death in 1 (0.3%) and MACCE in 3 (0.8%) in asymptomatic patients. Totally, stroke occurred in 3 (0.8%), intracranial hemorrhage in 5 (1.3%), death in 3 (0.8%), and MACCE in 11 (2.89%) patients.Long-term follow-up (40.95 ± 15.81 months) was performed on 295 (77.6%) patients; the results demonstrated a mortality rate of 13.9% (n = 41) which 30 (10.1%) cases belonged to symptomatic and 11 (3.7%) cases were from asymptomatic patients. Stroke happened in 26 (8.8%), that 18 (6.1%) cases were symptomatic and 8 (2.7%) cases were asymptomatic. MACCE rate was 19.3% (n = 57) which 41 (13.8%) cases were symptomatic and 16 (5.4%) were asymptomatic. Age (OR: 1.040, 95% CI: 1.002 to 1.079) was confirmed as a meaningful variable for the occurrence of MACCE. Conclusions: Our results confirm the safety and efficacy of carotid stenting in experienced centers. Age was as an important predictor of all-cause mortality, myocardial infarction, and MACCE.

Background and Objective: Discarded as wastes, parts of the agricultural products can be used for feed productivity as well as management of animal feed production. Production of various products is possible using appropriate processing. The objective of the present study was to use laccase of Albifimbria viridis in degradation of agricultural residues and to produce compounds with herbicide properties. Material and Methods: The fungi were isolated from agricultural soils. The isolates were identified using morphological detection and PCR amplification of the internal transcribed spacer. Supernatants were collected from semi-solid cultures and laccase activity was assessed using 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) substrate. This was carried out using n-hexane and degradation of the agricultural residues was investigated using gas chromatography-mass spectrometry. Results and Conclusion: Growth of the fungal isolate in culture media with tannic acid was studied using scanning electron microscopy. In total, the isolate produced 50 U ml-1 laccase. Gas chromatography-mass spectrometry analyses revealed production of oxime, methoxy-phenyl and 2-cyclopenten-1-one for tannic acids, o-guaiacol, tetradecane, hexadecane, octadecane, octadecanoic acid, hexadecanoic acid and benzene, 1,3-bis(1,1-dimethylethyl) for sorghum seeds and 2-acetyl-5-methylfuran, phenol, 2-methoxy and benzene, 1,2-dimethoxy for wheat straw during fungal growth (0.73 mg ml-1). Results have shown that the laccase enzyme produced from Albifimbria viridis native strain is capable of hydrolytic cleavage of chemical pollutants from agricultural wastes for herbicide bioremediation. Conflict of interest: The authors declare no conflict of interest.

Carbonic anhydrases (CAs) are metalloenzymes that can help organisms survive in hydrothermal vents by hydrating carbon dioxide (CO2). In this study, we focus on alpha (α), beta (β), and gamma (γ) CAs, which are present in the thermophilic microbiome of marine hydrothermal vents. The coding genes of these enzymes can be transferred between hydrothermal-vent organisms via horizontal gene transfer (HGT), which is an important tool in natural biodiversity. We performed big data mining and bioinformatics studies on α-, β-, and γ-CA coding genes from the thermophilic microbiome of marine hydrothermal vents. The results showed a reasonable association between thermostable α-, β-, and γ-CAs in the microbial population of the hydrothermal vents. This relationship could be due to HGT. We found evidence of HGT of α- and β-CAs between Cycloclasticus sp., a symbiont of Bathymodiolus heckerae, and an endosymbiont of Riftia pachyptila via Integrons. Conversely, HGT of β-CA genes from the endosymbiont Tevnia jerichonana to the endosymbiont Riftia pachyptila was detected. In addition, Hydrogenovibrio crunogenus SP-41 contains a β-CA gene on genomic islands (GIs). This gene can be transferred by HGT to Hydrogenovibrio sp. MA2-6, a methanotrophic endosymbiont of Bathymodiolus azoricus, and a methanotrophic endosymbiont of Bathymodiolus puteoserpentis. The endosymbiont of R. pachyptila has a γ-CA gene in the genome. If α- and β-CA coding genes have been derived from other microorganisms, such as endosymbionts of T. jerichonana and Cycloclasticus sp. as the endosymbiont of B. heckerae, through HGT, the theory of the necessity of thermostable CA enzymes for survival in the extreme ecosystem of hydrothermal vents is suggested and helps the conservation of microbiome natural diversity in hydrothermal vents. These harsh ecosystems, with their integral players, such as HGT and endosymbionts, significantly impact the enrichment of life on Earth and the carbon cycle in the ocean.

HNF4α, a member of the nuclear receptor superfamily, regulates the genes involved in lipid and glucose metabolism. The expression of the RARβ gene in the liver of HNF4α knock-out mice was higher versus wildtype controls, whereas oppositely, RARβ promoter activity was 50% reduced by the overexpression of HNF4α in HepG2 cells, and treatment with retinoic acid (RA), a major metabolite of vitamin A, increased RARβ promoter activity 15-fold. The human RARβ2 promoter contains two DR5 and one DR8 binding motifs, as RA response elements (RARE) proximal to the transcription start site. While DR5 RARE1 was previously reported to be responsive to RARs but not to other nuclear receptors, we show here that mutation in DR5 RARE2 suppresses the promoter response to HNF4α and RARα/RXRα. Mutational analysis of ligand-binding pocket amino acids shown to be critical for fatty acid (FA) binding indicated that RA may interfere with interactions of FA carboxylic acid headgroups with side chains of S190 and R235, and the aliphatic group with I355. These results could explain the partial suppression of HNF4α transcriptional activation toward gene promoters that lack RARE, including APOC3 and CYP2C9, while conversely, HNF4α may bind to RARE sequences in the promoter of the genes such as CYP26A1 and RARβ, activating these genes in the presence of RA. Thus, RA could act as either an antagonist towards HNF4α in genes lacking RAREs, or as an agonist for RARE-containing genes. Overall, RA may interfere with the function of HNF4α and deregulate HNF4α targets genes, including the genes important for lipid and glucose metabolism.

Background: Conventional laboratory culture-based methods for diagnosis of Streptococcus pneumoniae are time-consuming and yield false negative results. Molecular methods including real-time (RT)-PCR rapid methods and conventional PCR due to higher sensitivity and accuracy have replaced traditional culture assay. The aim of the current study was to evaluate lytA gene for the detection of Streptococcus pneumoniae in the cerebrospinal fluid of human patients with meningitis using real-time PCR assay. Material and Methods: In this cross-sectional study, a total of 30 clinical specimens were collected from patients in a period from September to December 2018. In order to evaluate the presence of lytA gene, conventional and real-time PCR methods were used without culture. Results: From 30 sputum samples, five (16.66%) isolates were identified as S. pneumoniae by lytA PCR and sequencing. Conclusion: In this research, an accurate and rapid real-time PCR method was used, which is based on lytA gene for diagnosis of bacteria. Based on the sequencing results, the sensitivity for the detection of lytA gene was 100% (5/5).

Bacillus spp. are the main sources of subtilisin E, which has several applications in biotechnology. The 3D structure of subtilisin E has a significant impact on its efficacy. In this study, we evaluated subtilisin E from Bacillus subtilis subsp. subtilis str. 168 by bioinformatic methods. The results revealed that the subtilisin E sequence from B. subtilis contains highly conserved amino acids, including histidine (H), aspartic acid (D) and serine (S). Subtilisin E cleaves the bonds between hydrophobic and polar amino acids in keratin-associated proteins. The effects of point mutations on the crystal structure of subtilisin E (PDB ID: 1SCJ) showed that changes of asparagine 123 (N123) to valine (V) and serine 331 (S331) to leucine (L) respectively, were the most stabilizing. Genomic analysis of the subtilisin E-coding gene (aprE) indicated that this gene and the yhfN gene are expressed through a σA promoter. The analysis of TBFs revealed AbrB, ScoC, DegU, Hpr, σA, SinR, TenA, and DegU as relevant regulators of aprE expression. Phylogenetic analysis showed that subtilisin Es have highly conserved structures among Bacillus spp., sharing a common ancestor, where their coding genes were duplicated and evolved within the Bacillus spp. As the conclusion, our in silico study demonstrated that the overexpression of the aprE gene and stability of the produced subtilisin E can be improved though system biology methods such as point mutations and identifying the involved transcription factors (TFs) or/and TBFs.Communicated by Ramaswamy H. Sarma.

Background: Canine babesiosis is one of the mainly worldwide-distributed tick-borne haemoprotozoan parasitic dis-eases in dogs. Methods: A total of 43 blood samples were randomly collected from naturally infected dogs in seven villages from different geographical areas of Meshkin Shahr, Ardabil Province, Iran. The presence of Babesia species detected with standard methods including parasitological and gene sequencing techniques targeting the 18S rRNA gene. Results: Our results revealed that four dogs 9.3% (4/43) including one female and three male dogs were infected with Babesia. All four Babesia-infected dogs were confirmed B. canis by the molecular-based method. Sequence alignments comparison of the B. canis genotypes A and B, it was revealed that all B. canis isolates belonged to genotype B. Conclusion: This study provides essential data for subsequently define the critical importance of the molecular studies in management and prevention of the canine babesiosis in Iran.

Study aim: In this study, we evaluated the effects of acute and chronic exercise on the plasma FAs and their association with cardiac hypertrophy indices. Material and methods: In this pilot study, 15 sedentary and 15 athlete women underwent acute and long-term water aerobic exercise and their plasma FA levels and a number of electrocardiographic parameters, such as left ventricular end-diastolic diameter index (LVEDDI), left ventricular ejection fraction (LVEF), left ventricular mass index (LVMI), and wall thickness were evaluated before and after the exercise program. Results: The acute exercise significantly increased palmitic and oleic acid levels in non-athletes and stearic acid in both groups. However, the same type of exercise decreased linoleic acid only in non-athlete women (p < 0.05). The water aerobics training caused a significant decrease in the levels of palmitic, stearic, and arachidonic acid, SFA/UFA, and ω3/ ω6 ratios and also an increase in α-Linolenic acid and MUFA in non-athletes. We found positive and negative correlations between LVEF with ω3 and SFA/UFA ratio in both groups, respectively. In the non-athlete group, the ω3/ω6 ratio showed negative correlations with LVMI and LVEDDI. Conclusions: The study indicated that the 12-week exercise by sedentary women could make their plasma FAs composition similar to athlete women. Moreover, the plasma FA levels were associated with cardiac hypertrophy indices, showing the importance of FAs in physiological hypertrophy.

Objective This study describes the occurrence of a silent mutation in the RNA binding domain of nucleocapsid phosphoprotein (N protein) coding gene from SARS-CoV-2 that may consequence to a missense mutation by onset of another single nucleotide mutation. Results In the DNA sequence isolated from severe acute respiratory syndrome (SARS-CoV-2) in Iran, a coding sequence for the RNA binding domain of N protein was detected. The comparison of Chinese and Iranian DNA sequences displayed that a thymine (T) was mutated to cytosine (C), so “TTG” from China was changed to “CTG” in Iran. Both DNA sequences from Iran and China have been encoded for leucine. In addition, the second T in “CTG” in the DNA or uracil (U) in “CUG” in the RNA sequences from Iran can be mutated to another C by a missense mutation resulting from thymine DNA glycosylase (TDG) of human and base excision repair mechanism to produce “CCG” encoding for proline, which consequently may increase the affinity of the RNA binding domain of N protein to viral RNA and improve the transcription rate, pathogenicity, evasion from human immunity system, spreading in the human body, and risk of human-to-human transmission rate of SARS-CoV-2.

Background: the aim of this study was the isolation of phages able to lyse some strains of MDR-K. pneumoniae (named vB_Kp1 and vB_Kp2) and E. aerogenes (named vB_Ea1) from swages. Materials and Methods: Different Klebsiella pneumoniae and Enterobacter aerogenesis strains isolated from clinical specimens (January to September 2018) in three Hospitals in Amol, (Mazandaran, Iran). Kirby Bauer’s disc diffusion method was used for determination of resistance profiles of these isolates using different antibiotics. The resistant strains to multi tested antibiotics (MDR) were selected to investigate the effect of isolated phages from wastewater and hospital sewage. Presence of phage investigated by plaque formation and after enriching and concentrating the isolated bacteriophages and staining the samples, a transmitting electron microscope (TEM) was used to observe the morphology of the bacteriophages. Phage identification tests including host range and One-step growth were performed. Results: TEM analysis revealed that tree phages have an icosahedral capsid and long contractile tail. Therefore, they are a member of the Myoviridae family. Phages were able to lyse 14 (56%) of the 25 strains of multidrug-resistant bacteria isolated. The one-step growth curve showed large burst sizes and short latent times. Conclusions: The formation of clear plaques shows the high lyse power of phages, so they have good potential for further analysis for clinical use as a therapeutic agent in the future.

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.

Abstract We report the production and biochemical characterization of an α-carbonic anhydrase (LrhCA) from gram-positive probiotic bacteria Lactobacillus rhamnosus GG. CAs form a family of metalloenzymes that catalyze hydration of CO2/interconversion between CO2 and water to bicarbonate ions and protons. They are divided into eight independent gene families (α, β, γ, δ, ζ, η, θ, and ι). Interestingly, many pathogens have been identified with only β- and/or γ-CAs, which can be targeted with CA-specific inhibitors (CAIs) acting as anti-pathogen drugs. Since it is important to study the potential off-target effects of CAIs for both the human body and its commensal bacteria, we took L. rhamnosus GG as our study subject. To date, only a single α-CA has been identified in L. rhamnosus GG, which was successfully produced and biochemically characterized. LrhCA showed moderate catalytic activity with the following kinetic parameters: kcat of 9.86 × 105 s−1 and kcat/KM of 1.41 × 107 s−1 M−1. Moderate inhibition was established with 11 of the 39 studied sulfonamides. The best inhibitors were 5-((4-aminophenyl)sulfonamido)-1,3,4-thiadiazole-2-sulfonamide, 4-(2-hydroxymethyl-4-nitrophenyl-sulfonamidoethyl)-benzenesulfonamide, and benzolamide with Ki values of 319 nM, 378 nM, and 387 nM, respectively. The other compounds showed weaker inhibitory effects. The Ki of acetazolamide, a classical CAI, was 733 nM. In vitro experiments with acetazolamide showed that it had no significant effect on cell growth in L. rhamnosus GG culture. Several sulfonamides, including acetazolamide, are in use as clinical drugs, making their inhibition data highly relevant to avoid any adverse off-target effects towards the human body and its probiotic organisms. Key points • The α-carbonic anhydrase from Lactobacillus rhamnosus GG (LrhCA) is 24.3 kDa. • LrhCA has significant catalytic activity with a kcat of 9.9 × 105 s-1. • Acetazolamide resulted in a marginal inhibitory effect on cell growth.

Nowadays, natural compounds have generated innovative attention for metal removal and precipitation; here, a fungus was isolated from soil samples in tannin media for metal removal. The isolate was identified as ‘Albifimbria viridisʼ by morphology and amplification of specific sequence. The isolate produced high activities of laccase when induced with tannin. The cell-free supernatant with 50 U/ml laccase activities could remove nickel, cobalt, cadmium, and copper nitrate salts by 59.69%, 59.75%, 46.97%, and 66.79%, respectively, while the precipitation of crude enzyme and metal salts by Fourier transform infrared spectroscopy analyses showed that only 2 metal salts (Cobalt and Copper nitrate) are precipitated in oxide form. Tannin was used as a mediator for metal removal, and the data were supported using a zymogram with partially purified laccase and metal precipitation on polyacrylamide gel electrophoresis. A 66 KDa laccase can remove metals and tannin. The metal interaction by this isolated compared with Phanerochaete chrysosporium and Aspergillus niger. All studied fungi could biodegrade the tannin as sole carbon source; however, for maximum activities, the new isolate Albifimbria viridis was selected in further study. The purified fungus in this study is a herbicide and has a high activity of laccase. The precipitation of metals by the enzyme of grown fungus in tannic acid is reported for the first time, which promoted the functional importance of this fungus.

Background: The last two decades witnessed the spread of the first generation of influenza viruses. Influenza virosomes are promising tools in vaccine and immunotherapy programs because of their applications in various medical fields. The aim of the present study was to construct cationic virosomes derived from influenza virus using dialyzable detergent (DCPC) and cationic lipid (DOTAP) in vitro. Materials and Methods: Influenza A / Puerto Rico / 34.8 (H1N1) strain was propagated in MDCK cell line. The influenza virus envelope was dissolved using DCPC as a dialyzable detergent, and finally it was removed by dialysis and the cationic virosome was synthesized through adding cationic lipid (DOTAP). Cytotoxicity and presence of HA and NA proteins were evaluated by cell viability assay (MTT assay) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), respectively. Also, macroscopic and morphology studies of virosomes were performed by transmission electron microscopy (TEM). Results: The final concentration of virosomes was 1.5 mg/ml. The presence of HA and NA proteins was confirmed by SDS-PAGE. Cell survival was significantly decreased after 48 hours of treatment with different concentrations of cationic virosomes (P

In this study, a solvothermal method at the temperature of 180 °C for 4 h was used for the synthesis of hydroxyapatite-reduced graphene oxide (HA-rGO) nanopowders by employing the nitrogen gas injection. To synthesize the powders, a solvent containing dimethylformamide, anhydrous ethanol, diethylene glycol, and water was used (1:1:1:2, respectively). Calcium nitrate tetrahydrate and diammonium hydrogenphosphate were used as calcium and phosphate precursors, respectively. The synthesized powders were consolidated by a high-frequency induction heat sintering method. The powders and sintered samples were then evaluated using X-ray diffraction, Raman spectroscopy, high-resolution transmission electron microscopy, Vickers indentation technique, and biocompatibility assay. The findings of this study showed that the final powders synthesized by the solvothermal method had calcium to phosphate ratio of about 1.67. The high frequency induction heat sintering method was suitable for consolidating these powders and exhibited a higher relative density than other methods. The mechanical properties of the nanocomposite were higher than those of pure HA. In biological experiments the difference of HA effect in comparison with the nanocomposite was not significant and both were biocompatible.

Background & Aims of the Study: Selenium is an essential nutritional material used for the important functions of the human body. The issue of the production of selenium nanoparticles (SeNPs) was investigated in various fields, such as anticancer, antioxidant, and antibacterial activities. Materials and Methods: In order to study antibacterial activity, the nutrient broth medium containing synthesized SeNPs in six different concentrations (i.e., 100, 50, 25, and 12.5 µM) was evaluated on six pathogenic bacteria. Then, the growth curves were drawn as an antibacterial assay in six pathogenic bacteria. In addition, the antioxidant effect was examined by the 2, 2-Diphenyl-1-picrylhydrazyl method. The stock solution of SeNPs was mixed with culture media in six different concentrations (i.e., 0.001, 0.01, 0.1, 1, 10, and 100 μM) and exposed on MCF-7 and HT-29 cell lines; therefore, the anticancer effects of SeNPs were assayed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. Results: According to the obtained results, the most and least significant antibacterial effects of synthesized SeNPs were observed for Staphylococcus aureus (98%) and Klebsiella pneumonia (51.55%), respectively. The highest concentration of biosynthesized SeNPs can achieve a better outcome regarding the antioxidant activity. The growth of MCF-7 and HT-29 cancer cell lines were inhibited by synthesized SeNPs in a concentration of 100 µM. Conclusion: Consequently, the results of the present study showed that SeNPs synthesized from indigenous halophilic bacteria could display anticancer, antioxidant, and antibacterial activities. This progress can assist in the treatment of different diseases.

Background Mycoplasma pneumoniae is one of the widespread causes of community-acquired pneumonia (CAP). Over recent years, the widespread use of macrolides has led to the emergence of macrolide-resistant M.pneumoniae (MRMP) resulted from mutations at specific positions of domain V of the 23S rRNA gene. Methods We collected 100 samples of throat swabs from patients with respiratory infections. After extraction of DNA from bacterial cell cultured in PPLO broth media using Roche kit (Germany), the PCR was performed on specific samples of M. pneumoniae using specific primers for 23S rRNA gene.Afterwards, for positive samples, minimum inhibitory concentration (MIC) was determined using the broth microdilution with Clarithromycin. Finally, the PCR product was sequenced to detect mutations related to macrolide resistance in domain V of 23S rRNA . Results According to the analysis of the sequenced PCR product of M. pneumoniae 23S rRNA gene using Clustalw2 online software, one of the samples were shown to have a mutation at A2431G and G2491A positions. The MIC measurement also revealed that all isolates were sensitive to Clarithromycin, and there was no macrolide resistance to Clarithromycin in all isolates. Conclusion Sequence analysis of the 23S rRNA gene in M. pneumoniae , revealed no macrolide resistance of M. pneumoniae to Clarithromycin. Thus, the use of these antibiotics should be restricted to prevent the development of macrolide-resistant M. pneumoniae in Iran.

Background: Microorganisms attach to various surfaces and they have manufactured biofilms by production polysaccharides like PSL in P.aeruginosa. synthesis of this kind of polysaccharide has done by PSL gene cluster. The aim of this study is consideration biofilm formation which is one of the major cause antibiotic resistance Methods: In this study, 100 P. aeruginosa were isolated with bacteriological and biochemical methods and pslA gene detection with PCR in all of the P. aeruginosa isolated from patients Then biofilm formation checked with microtiter plate method and it showed with SEM. Finally, expression of main attachment gene pslA in 6 strains could make moderate and strong biofilm were investigated by real-time PCR assay.Results:In this study, 100 P. aeruginosa were isolated that these strains showed High rates of MDR. The presence pslA gene in all of the pseudomonas isolated from patients was proven. Microtiter plate method showed 24 (24%) strains could make biofilm Among 100 strains that showed with SEM. The pslA expression in strains which making moderate and strong biofilms are more than other strains Conclusions: Hence, for bacterial biofilm treatment is recommended: Before antibiotics are prescribed, biofilm formation by bacteria should be investigated.

Background: The prevalence of carbapenem-resistant Enterobacteriaceae, especially Klebsiella pneumoniae carbapenemase (KPC), has been recently reported worldwide. Therefore, there is an indispensable need for precise and rapid detection of these carbapenemases. Objectives: This study aimed to propose an accurate and rapid method for detecting K. pneumoniae carbapenemase genes from clinical samples, using reverse transcription-polymerase chain reaction (RT-PCR) and to evaluate the expression of these genes in the presence of β-lactam antibiotic by real-time PCR assay.Methods: One hundred and eighty-one K. pneumoniae strains were collected from patients presenting to Firoozgar Hospital of Tehran, Iran. The strains were tested using the disk diffusion method, the modified Hodge test (MHT), and E-test minimum inhibitory concentration (MIC). Next, reverse transcription-PCR method was applied for the identification of bla OXA-23 and bla OXA-48 genes. Finally, expression of genes was measured by real-time PCR assay in the presence and absence of β-lactam antibiotic.Results: Phenotypic testing also showed a high level of antibiotic resistance, while the genotypic methods indicated the presence and expression of carbapenemase genes.Conclusions: The findings suggest revisions in the current antibiotic therapy protocols, considering the high expression level of resistant carbapenemases to K. pneumoniae strains.

In this study, hydroxyapatite-reduced graphene oxide (HA-rGO) powders were first synthesized in situ using a hydrothermal method. These powders were then consolidated using a cold sintering method. The solvent used in this method was water + dimethylformamide + brushite which was added to the powders at different ratios. The sintered samples were then evaluated using X-ray diffraction, Fourier transform infrared spectroscopy, Raman spectroscopy, high-resolution transmission electron microscopy, and Vickers microindentation. The results of this study showed that the best conditions for the sintering of rGO-HA nanopowders were a temperature of 200 °C, a holding time of >30 min, and a pressure of 500 MPa. The best mechanical properties were achieved when the solvent content was considered to be 20 wt%. Crack deflection and graphene bridging were among the mechanisms that increased the fracture toughness of these nanocomposites. By adding 1.5% rGO, the fracture toughness of this nanocomposite (using the cold sintering method) was approximately equivalent to that of spark plasma sintered HA.

We report the biochemical and structural characterisation of a beta-carbonic anhydrase (β-CA) from Trichomonas vaginalis, a unicellular parasite responsible for one of the world’s leading sexually transmitted infections, trichomoniasis. CAs are ubiquitous metalloenzymes belonging to eight evolutionarily divergent groups (α, β, γ, δ, ζ, η, θ, and ι); humans express only α-CAs, whereas many clinically significant pathogens express only β- and/or γ-CAs. For this reason, the latter two groups of CAs are promising biomedical targets for novel antiinfective agents. The β-CA from T. vaginalis (TvaCA1) was recombinantly produced and biochemically characterised. The crystal structure was determined, revealing the canonical dimeric fold of β-CAs and the main features of the enzyme active site. The comparison with the active site of human CA enzymes revealed significant differences that can be exploited for the design of inhibitors selective for the protozoan enzyme with respect to the human ones.

Background: The influenza virus's Nucleoprotein (NP) is a highly conserved protein used in vaccine design. Alum, besides its neurotoxicity, is a commonly used adjuvant. Objectives: We tested the NP's efficacy as an alum-free flu vaccine. Methods: The recombinant vector (NP-pET-28a) was expressed in a prokaryotic system. Then, the immunogenicity of NP, alone or with alum, was evaluated in Balb/C mice by measuring IgGs (total and subtypes), interferon-gamma (IFN-γ), and interleukin-4 (IL-4) production and animal surveillance. Results: NP+Alum has 71.4% animal surveillance, whereas NP had 57.1%. While NP+Alum produced considerable IgGs, IFN-, and IL-4, NP alone showed promising effects. Conclusion: The NP alone may give adequate protection against the H1N1 strain of Influenza A.

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.The mutation screening was done by whole exome sequencing followed by direct Sanger sequencing.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.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.

There are several human viruses with known potential for causing cancers including, Hepatitis B virus, Hepatitis C virus, Epstein-Barr virus, Kaposi's sarcoma herpesvirus, Human T-cell lymphotropic virus, Human papillomavirus, and Merkel cell polyomavirus. Cancer is the second leading cause of death that affects humans worldwide, especially in developing countries. Surgery, chemotherapy, and radiotherapy can cure about 60% of humans with cancer but recurrent and metastatic diseases remain a major reason for death. In recent years, understanding the molecular characteristics of cancer cells has led to the improvement of therapeutic strategies using novel emerging therapies. Oncolytic viruses with the potential of lysing cancer cells defined the field of oncolytic virology, hence becoming a biotechnology tool rather than just a cause of disease. This study mainly focused on targeting cell proliferation and death pathways in human tumor-inducing viruses by developing innovative therapies for cancer patients based on the natural oncolytic properties of reovirus. To kill tumor cells efficiently and reduce the chance of recurrence both the direct ability of reovirus infection to lyse the tumor cells and the stimulation of a potent host immune response are applied. Hence, bioengineered stem cells can be used as smart carriers to improve the efficacy of oncolytic reovirus and safety profiles.