Combination of microRNA and suicide gene for targeting Glioblastoma: Inducing apoptosis and significantly suppressing tumor growth in vivo.

Glioblastoma (GBM), a grade IV brain tumor, presents a severe challenge in treatment and eradication due to its high genetic variability and the existence of stem-like cells with self-renewal potential. Conventional therapies fall short of preventing recurrence and fail to extend the median survival of patients significantly. However, the emergence of gene therapy, which has recently obtained significant clinical outcomes, brings hope. It has the potential to be a suitable strategy for the treatment of GBM. Notably, microRNAs (miRNAs) have been noticed as critical players in the development and progress of GBM. The combined usage of hsa-miR-34a and Cytosine Deaminase (CD) suicide gene and 5-fluorocytosine (5FC) prodrug caused cytotoxicity against U87MG Glioma cells in vitro. The apoptosis and cell cycle arrest rates were measured by flow cytometry. The lentiviral vector generated overexpression of CD/miR-34a in the presence of 5FC significantly promoted apoptosis and caused cell cycle arrest in U87MG cells. The expression level of the BCL2, SOX2, and P53 genes, target genes of hsa-miR-34a, was examined by quantitative real-time PCR. The treatment led to a substantial downregulation of Bcl2 and SOX2 genes while elevating the expression levels of Caspase7 and P53 genes compared to the scrambled control. The hsa-miR-34a hindered the proliferation of GBM cancer cells and elevated apoptosis through the P53-miR-34a-Bcl2 axis. The CD suicide gene with 5FC treatment demonstrated similar results to miR-34a in the apoptosis, cell cycle, and real-time assays. The combination of CD and miR-34a produced a synergistic effect. In vivo, anti-GBM efficacy evaluation in rats bearing intracranial C6 Glioma cells revealed a remarkable induction of apoptosis and a significant inhibition of tumor growth compared with the scrambled control. The simultaneous use of CD/miR-34a with 5FC almost entirely suppressed tumor growth in rat models. The combined application of hsa-miR-34a and CD suicide gene against GBM tumors led to significant induction of apoptosis in U87MG cells and a considerable reduction in tumor growth in vivo.

The impact of host microRNAs on the development of conserved mutations of SARS-CoV-2.

SARS-CoV-2, the virus responsible for the COVID-19 pandemic, has undergone various genetic alterations due to evolutionary pressures exerted by host cells, including intracellular antiviral mechanisms such as targeting by human microRNAs (miRNAs). This study investigates the impact of miRNAs hsa-miR-3132 and hsa-miR-4650 on the viral genome. Sequence alignment revealed conserved mutations in the binding sites of these miRNAs in adapted strains compared to the original Wuhan-Hu-1 strain, leading to their deletion. Despite modest expression of these miRNAs in SARS-CoV-2 target tissues, their efficacy against mutant strains is reduced due to the loss of binding sites. Structural analysis indicates that the mutant genome is more stable than the Wuhan-Hu-1 genome. Luciferase and virus titration assays demonstrate that hsa-miR-3132 and hsa-miR-4650 effectively target the Nsp3 gene in the Wuhan-Hu-1 strain but not in mutant strains lacking their binding sites. These findings suggest that the observed mutations help the virus evade selective pressure from human miRNAs, contributing to its adaptation.

Activation of the BMP2/SMAD4 signaling pathway for enhancing articular cartilage regeneration of mesenchymal stem cells utilizing chitosan/alginate nanoparticles on 3D extracellular matrix scaffold.

This study investigated the efficacy of using chitosan/alginate nanoparticles loaded with recombinant human bone morphogenetic-2 (rhBMP-2) and SMAD4 encoding plasmid to enhance the chondrogenesis of human bone marrow mesenchymal stem cells (hBM-MSCs) seeded on an extracellular matrix (ECM). The research treatments included the stem cells treated with the biological cocktail (BC), negative control (NC), hBM-MSCs with chondrogenic medium (MCM), hBM-MSCs with naked rhBMP-2 and chondrogenic medium (NB/C), and hBM-MSCs with naked rhBMP-2 and chondrogenic medium plus SMAD4 encoding plasmid transfected with polyethyleneimine (PEI) (NB/C/S/P). The cartilage differentiation was performed with real-time quantitative PCR analysis and alizarin blue staining. The data indicated that the biological cocktail (BC) exhibited significantly higher expression of cartilage-related genes compared to significant differences with MCM and negative control (NC) on chondrogenesis. In the (NB/C/S/P), the expression levels of SOX9 and COLX were lower than those in the BC group. The expression pattern of the ACAN gene was similar to COL2A1 changes suggesting that it holds promising potential for cartilage regeneration.

Chitosan-Aloe Vera Composition Loaded with Zinc Oxide Nanoparticles for Wound Healing: In Vitro and In Vivo Evaluations.

Global concerns due to the negative impacts of untreatable wounds, as well as the growing population of these patients, emphasize the critical need for advancements in the wound healing materials and techniques. Nanotechnology offers encouraging avenues for improving wound healing process. In this context, nanoparticles (NPs) and certain natural materials, including chitosan (CS) and aloe vera (AV), have demonstrated the potential to promote healing effects. The objective of this investigation is to assess the effect of novel fabricated nanocomposite gel containing CS, AV, and zinc oxide NPs (ZnO NPs) on the wound healing process. The ZnO NPs were synthesized and characterized by X-ray diffraction and electron microscopy. Then, CS/AV gel with different ratios was prepared and loaded with ZnO NPs. The obtained formulations were characterized in vitro based on an antimicrobial study, and the best formulations were used for the animal study to assess their wound healing effects in 21 days. The ZnO NPs were produced with an average 33 nm particle size and exhibited rod shape morphology. Prepared gels were homogenous with good spreadability, and CS/AV/ZnO NPs formulations showed higher antimicrobial effects against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. The wound healing findings showed significant wound area reduction in the CS/AV/ZnO NPs group compared to negative control at day 21. Histopathological assessment revealed the advantageous impact of this formulation across various stages of the wound healing process, including collagen deposition (CS/AV/ZnO NPs (2 : 1), 76.6 ± 3.3 compared to negative control, 46.2 ± 3.7) and epitheliogenesis (CS/AV/ZnO NPs (2 : 1), 3 ± 0.9 compared to negative control, 0.8 ± 0.8). CS/AV gel-loaded ZnO NPs showed significant effectiveness in wound healing and would be suggested as a promising formulation in the wound healing process. Further assessments are warranted to ensure the robustness of our findings.

Inhibitory effect of miR342 on the progression of triple-negative breast cancer cells in vitro and in the mice model.

Introduction: Breast cancer is the most common cancer in women worldwide, and the triple-negative subtype is the most invasive, with limited therapeutic options. Since miRNAs are involved in many cellular processes, they harbor great value for cancer treatment. Therefore, in this study, we have investigated the anti-proliferative and anti-invasive roles of miR342 in 4T1 triple-negative cells in vitro and also studied the effect of this miRNA on tumor progression and the expression of its target genes in vivo. Methods: 4T1 cells were transduced with conditioned media of miR342-transfected Hek-LentiX cells. MTT and clonogenic assays were used to assess the viability and colony-forming ability of 4T1 cells. Apoptosis and invasion rates were respectively evaluated by annexin/7-AAD and wound healing assays. At last, in vivo tumor progression was evaluated using H&E staining, real-time PCR, and immunohistochemistry. Results: The viability of transduced-4T1 cells reduced significantly 48 hours after cell seeding and colony forming ability of these cells reduced to 50% of the control group. Also, miR342 imposed apoptotic and anti-invasive influence on these cells in vitro. A 30-day follow-up of the breast tumor in the mice model certified significant growth suppression along with reduced mitotic index and tumor grade in the treatment group. Moreover, decreased expression of Bcl2l1, Mcl1, and ID4, as miR342 target genes, was observed, accompanied by reduced expression of VEGF and Bcl2/Bax ratio at the protein level. Conclusion: To conclude, our data support the idea that miR342 might be a potential therapeutic target for the treatment of triple-negative breast cancer (TNBC).

Mining the soil myxobacteria and finding sources of anti-diabetic metabolites.

Secondary metabolites produced by myxobacterial genera are often characterized as diverse molecules with unique structural properties which drove us to search for myxobacterial source of anti-diabetic drug discovery. In the present study, from 80 soil samples, out of sixty-five observed isolates, 30 and 16 were purified as Myxococcus and non-Myxococcus, respectively. Isolated strains taxonomically belonged to the genera Myxococcus, Corallococcus and Cystobacter, Archangium, Nanocystis, and Sorangium, and some could not be attributed. Secondary metabolites of selected non-Myxococcus isolates extracted by the liquid-liquid method showed that the myxobacterium UTMC 4530 demonstrated the highest inhibition on the formation of carbonyl group and fructosamine, respectively. In addition, it showed 23% and 15.8% inhibitory activity on α-glucosides and α-amylase compared to acarbose (23%, 18%), respectively. The extract of strain UTMC 4530 showed 35% induction effect on glucose adsorption while showing no radical scavenging activity and no toxic effect on HRBC lysis and HepG2 in cytotoxicity assays. The strain UTMC 4530 (ON808962), with the multiple antidiabetic activity, showed 87.3% similarity to Corallococcus llansteffanensis which indicates its affiliation to a new genus. The results of this study revealed that secondary metabolites produced by strain UTMC 4530 can be considered a promising source to find new therapeutic and pharmaceutical applications perhaps a multi-mechanism anti-diabetic compound.

Augmented antiviral activity of chlorhexidine gluconate on herpes simplex virus type 1, H1N1 influenza A virus, and adenovirus in combination with salicylic acid.

BACKGROUND: The excessive usage of chlorhexidine gluconate (CHG) as a broad-spectrum antimicrobial reagent can have a negative impact on the environment and on human health. The aim of this study was to investigate the effectiveness of some plant-derived compounds in reducing the CHG concentration required to exert its antiviral activity. METHODS: Antiviral assays were conducted according to EN 14476 (2019) against herpes simplex virus type 1 (HSV-1), H1N1 influenza A virus, and adenovirus type 5 (Ad-5) as enveloped and non-enveloped viral models to assess the synergistic interaction of CHG and natural additive compounds. RESULTS: The effective concentration of 0.247 mM CHG against HSV-1 was decreased tenfold in combination with 0.0125 mM salicylic acid, with a titer reduction of 1.47 ⋅ 104 CCID50/ml. The time required for complete inactivation of HSV-1 particles was reduced to 15 min when the virus was exposed to 0.061 mM CHG and 0.0125 mM salicylic acid. Additionally, the presence of salicylic acid protected the CHG activity against interfering substances. CONCLUSION: Our supplemented CHG formulation showed immediate antiviral effectiveness, which is important for management of the infections. CHG can be combined with salicylic acid to exhibit synergistic antiviral activity at lower concentrations and reduce the time required for inactivation. Furthermore, in the presence of interfering substances, the combination has higher antiviral activity than CHG alone.

The role of microRNAs in the pathophysiology of human central nervous system: A focus on neurodegenerative diseases.

microRNAs (miRNAs) are suggested to play substantial roles in regulating the development and various physiologic functions of the central nervous system (CNS). These include neurogenesis, cell fate and differentiation, morphogenesis, formation of dendrites, and targeting non-neural mRNAs. Notably, deregulation of an increasing number of miRNAs is associated with several neurodegenerative diseases including Alzheimer's disease, Parkinson's disease, multiple sclerosis, amyotrophic lateral sclerosis and CNS tumors. They are particularly known to affect the amyloid ß (Aß) cleavage and accumulation, tau protein homeostasis, and expression of alpha-synuclein (α-syn), Parkin, PINK1, and brain-derived neurotrophic factor (BDNF) that play pivotal roles in the pathogenesis of neurodegenerative diseases. These include miR-16, miR-17-5p, miR-20a, miR-106a, miR-106b, miR-15a, miR-15b, miR-103, miR-107, miR-298, miR-328, miR-195, miR-485, and miR-29. In CNS tumors, several miRNAs, including miR-31, miR-16, and miR-21 have been identified to modulate tumorigenesis through impacting tumor invasion and apoptosis. In this review article, we have a look at the recent advances on our knowledge about the role of miRNAs in human brain development and functions, neurodegenerative diseases, and their clinical potentials.

The role of microRNA-185 in the pathogenesis of human diseases: A focus on cancer.

MicroRNAs (miRNAs) are a widely-studied class of non-coding RNAs characterized by their short length (18-25 nucleotides). The precise functions of miRNAs are not well-elucidated; however, an increasing number of studies suggest their involvement in various physiologic processes and deregulation in pathologic conditions. miRNA-185 (miR-185) is among the mostly-studied miRNAs in human diseases, which is found to play putative roles in conditions like metabolic disorders, asthma, frailty, schizophrenia, and hepatitis. Notably, many cancer studies report the downregulation of miR-185 in cell lines, tumor tissues, and plasma specimens of patients, while it demonstrates a suppressing role on the malignant properties of cancer cells in vitro and in vivo. Accordingly, miR-185 can be considered a tumor suppressor miRNA in human malignancies, while a few studies also report inconsistent findings. Being suggested as a prognostic/diagnostic biomarker, mi-185 is also found to offer clinical potentials, particularly for early diagnosis and prediction of the prognosis of cancer patients. In this review, we have outlined the studies that have evaluated the functions and clinical significance of miR-185 in different human diseases with a particular focus on cancer.

microRNA-382 as a tumor suppressor? Roles in tumorigenesis and clinical significance.

MicroRNAs (miRNAs) are small single-stranded RNAs belonging to a class of non-coding RNAs with an average length of 18-22 nucleotides. Although not able to encode any protein, miRNAs are vastly studied and found to play role in various human physiologic as well as pathological conditions. A huge number of miRNAs have been identified in human cells whose expression is straightly regulated with crucial biological functions, while this number is constantly increasing. miRNAs are particularly studied in cancers, where they either can act with oncogenic function (oncomiRs) or tumor-suppressors role (referred as tumor-suppressor/oncorepressor miRNAs). miR-382 is a well-studied miRNA, which is revealed to play regulatory roles in physiological processes like osteogenic differentiation, hematopoietic stem cell differentiation and normal hematopoiesis, and liver progenitor cell differentiation. Notably, miR-382 deregulation is reported in pathologic conditions, such as renal fibrosis, muscular dystrophies, Rett syndrome, epidural fibrosis, atrial fibrillation, amelogenesis imperfecta, oxidative stress, human immunodeficiency virus (HIV) replication, and various types of cancers. The majority of oncogenesis studies have claimed miR-382 downregulation in cancers and suppressor impact on malignant phenotype of cancer cells in vitro and in vivo, while a few studies suggest opposite findings. Given the putative role of this miRNA in regulation of oncogenesis, assessment of miR-382 expression is suggested in a several clinical investigations as a prognostic/diagnostic biomarker for cancer patients. In this review, we have an overview to recent studies evaluated the role of miR-382 in oncogenesis as well as its clinical potential.

Design, optimization and characterization of a novel antibacterial chitosan-based hydrogel dressing for promoting blood coagulation and full-thickness wound healing: A biochemical and biophysical study.

The use of hydrogel dressings has become increasingly popular as a scaffold for skin tissue engineering. Herein, we have developed an innovative wound dressing using chitosan, fibrinogen, nisin, and EDTA as an effective antibacterial scaffold for wound treatment. The structural and functional characteristics of the hydrogel, including morphology, mechanical strength, drug encapsulation and release, swelling behaviors, blood coagulation, cytotoxicity, and antibacterial activity, were studied. Spectroscopic studies indicated that the attachment of chitosan to fibrinogen is associated with minimal change in its secondary structure; subsequently, at higher temperatures, it is expected to preserve fibrinogen's conformational stability. Mechanical and blood coagulation analyses indicated that the incorporation of fibrinogen into the hydrogel resulted in accelerated clotting and enhanced mechanical properties. Our cell studies showed biocompatibility and non-toxicity of the hydrogel along with the promotion of cell migration. In addition, the prepared hydrogel indicated an antibacterial behavior against both Gram-positive and Gram-negative bacteria. Interestingly, the in vivo data revealed enhanced tissue regeneration and recovery within 17 days in the studied animals. Taken together, the results obtained from in vitro and histological assessments indicate that this innovatively designed hydrogel shows good potential as a candidate for wound healing.

The miR-429 suppresses proliferation and migration in glioblastoma cells and induces cell-cycle arrest and apoptosis via modulating several target genes of ERBB signaling pathway.

BACKGROUND: Glioblastoma multiforme (GBM) is an aggressive and lethal brain cancer, which is incurable with standard cancer treatments. miRNAs have great potential to be used for gene therapy due to their ability to modulate several target genes simultaneously. We found miR-429 is downregulated in GBM and has several predicted target genes from the ERBB signaling pathway using bioinformatics tools. ERBB is the most over-activated genetic pathway in GBM patients, which is responsible for augmented cell proliferation and migration in GBM. METHODS AND RESULTS: Here, miR-429 was overexpressed using lentiviral vectors in U-251 and U-87 GBM cells and it was observed that the expression level of several oncogenes of the ERBB pathway, EGFR, PIK3CA, PIK3CB, KRAS, and MYC significantly decreased, as shown by real-time PCR and western blotting. Using the luciferase assay, we showed that miR-429 directly targets MYC, BCL2, and EGFR. In comparison to scrambled control, miR-429 had a significant inhibitory effect on cell proliferation and migration as deduced from MTT and scratch wound assays and induced cell-cycle arrest and apoptosis in flow cytometry. CONCLUSIONS: Altogether, miR-429 seems to be an efficient suppressor of the ERBB genetic signaling pathway and a potential therapeutic for GBM.

Enhancement of bactericidal effect of Chlorhexidine using choline augmentation as a natural additive.

BACKGROUND: The long-term use of Chlorhexidine (CHG) at high concentrations has detrimental effects on health and the environment. This research was aimed to evaluate the antibacterial capacity of CHG in the presence of choline as a natural additive to reduce the amount CHG required for the activity. METHODS: The newly obtained formulation was evaluated for its stability and efficiency under physicochemical stresses. The checkerboard test was also performed to evaluate the type of CHG-additive interaction in a combinational state. RESULTS: The MIC of CHG was reduced up to five-fold in combination with choline at the concentration of 50mg/L and five log cfu/mL at 3 minutes of exposure. Chlorhexidine activity in combination with choline was preserved 20% more in the presence of interfering substances. The activity between CHG and choline was demonstrated as synergistic with FICI of 0.375. At sub MIC concentrations of both CHG (≤20 mg/L) and choline (≤6 mg/L) the observed interaction was synergy. While the significant interaction at high concentrations for both compounds was indifferent manner. The lysis effect of the CHG 20 mg/L was diminished in the presence of choline. CONCLUSIONS: Our suggested CHG formulation demonstrated rapid antimicrobial efficacy which can be a useful adjunct to infection control strategies currently employed in healthcare facilities. In conclusion, CHG can be combined with natural compounds as additives for enhanced synergistic antimicrobial activity.

The Potential Therapeutic Effect of RNA Interference and Natural Products on COVID-19: A Review of the Coronaviruses Infection.

The SARS-CoV-2 virus was reported for the first time in Wuhan, Hubei Province, China, and causes respiratory infection. This pandemic pneumonia killed about 1,437,835 people out of 61,308,161cases up to November 27, 2020. The disease's main clinical complications include fever, recurrent coughing, shortness of breath, acute respiratory syndrome, and failure of vital organs that could lead to death. It has been shown that natural compounds with antioxidant, anticancer, and antiviral activities and RNA interference agents could play an essential role in preventing or treating coronavirus infection by inhibiting the expression of crucial virus genes. This study aims to introduce a summary of coronavirus's genetic and morphological structure and determine the role of miRNAs, siRNAs, chemical drugs, and natural compounds in stimulating the immune system or inhibiting the virus's structural and non-structural genes that are essential for replication and infection of SARS-CoV-2.

Physicochemical susceptibility of SARS-CoV-2 to disinfection and physical approach of prophylaxis.

The transmission control of the newly emerged severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the most effective strategy by the absence of its specified vaccine or drug. Although the aerosol mediated transmission of SARS-CoV-2 has been confirmed, the physicochemical treatment of the biotic and abiotic objects is still the most promising approach in its infection control. The front line of the most effective disinfecting compounds on SARS-CoV-2 implies to be sodium hypochlorite, ethanol, hydrogen peroxide, quaternary ammonium compounds, and phenolic compounds, respectively. However, widely used compounds of alkyldimethylbenzylammonium chloride (benzalkonium chloride) biguanides (chlorhexidine) have not shown the multitude load reduction in less than 10 minutes. The susceptibility of SARS-CoV-2 to physical treatment follows the pattern of heat, acidity, and UV radiation. Rather all of the mentioned physical or chemical treatments, target the envelope proteins of the coronavirus mainly by impairing its entry to host cells. The anti-SARS-CoV-2 activity of combinatorial physicochemical treatments or evaluation of new chemical entities or physical treatments such as microwave irradiation still needs to be explored. Therefore, the development of a reliable decontamination protocol for SARS-CoV-2 demands revealing its stability pattern study vs a spectrum of single and combinatorial physicochemical parameters.

Bioprospecting of indigenous myxobacteria from Iran and potential of Cystobacter as a source of anti-MDR compounds.

Drug resistance is a critical issue in future clinical treatment. Methicillin-resistant Staphylococcus aureus (MRSA) is among the pathogens that need indispensable drug-discovery efforts. The myxobacteria are a unique group of bacteria that have recently been regarded for their potency to produce new drugs with high chemical diversity and unusual mode of actions. The present study was conducted to isolate and screen myxobacteria for the first time from Iran habitats and evaluate their antibacterial activity against the multidrug-resistant strain of S. aureus. Out of 62 soil and rotten plant samples, 51 myxobacteria were isolated. The isolates belonged to Myxococcus, Corallococcus, Pyxidicoccus, and Cystobacter genera based on morphology and 16S rRNA gene sequencing. Secondary metabolites of the selected strains were screened for activity on MDR strain with resistance to multiple antibiotic classes. The semi-purified fraction from Cystobacter sp. UTMC 4086 showed potent activity against MDR S. aureus with minimum inhibitory effect at 5 ≥ µg per mL compared with vancomycin (5 µg per mL) as well as no toxicity against Artemia salina. Hence, the strain Cystobacter sp. UTMC 4086 can be a valuable candidate for antibiotic discovery against MRSA and its metabolites can be subjected to further purification and analysis aimed at the identification of the effective chemical entity.