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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.

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.

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.

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 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.

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.

Leishmaniosis is caused by different species of Leishmania parasites. The available treatments for this disease have not provided strong consistent results yet. The weak response of current chemotherapeutics can be attributed to their deficient effects on stealth parasites inside macrophages, rapid clearance from the site of action, and systemic side effects in high doses. To enhance leishmaniosis vaccine efficacy, it is a valuable strategy to use liposomes as vaccine delivery systems due to combined increase in technological advances and understanding of the immune system. Liposomes that contain and deliver immunostimulators and antigens are now being developed to target diseases that require stimulation of both humoral and cell-mediated immune responses. Hence, using particulate adjuvants, like liposomes for effective delivery to the antigen presenting cells (APCs) is important for improving leishmaniosis vaccine efficacy. This study aimed at reviewing liposomal adjuvants in vaccine development with specific accentuation on their adjuvant mechanism and surface charge. It also examined how specific physicochemical qualities of liposomes and the particle size during formulation design can affect the immune response.