Gentamicin-loaded chitosan/folic acid-based carbon quantum dots nanocomposite hydrogel films as potential antimicrobial wound dressing.

BACKGROUND: To provide effective healing in the wound, various carbohydrate polymers are commonly utilized that are highly potent platforms as wound dressing films. In this work, novel antibacterial flexible polymeric hydrogel films were designed via crosslinking polymeric chitosan (CS) with folic acid-based carbon quantum dots (CQDs). To end this, folic acid as a bio-precursor is used to synthesize CQDs through the hydrothermal technique. The synthesized CQDs as a crosslinking agent was performed at different concentrations to construct nanocomposite hydrogel films via the casting technique. Also, gentamicin (GM), L-Arginine and glycerol were supplemented in the formulation of nanocomposite since their antibiotic, bioactivity and plasticizing ability, respectively. RESULTS: The successful construction of films were verified with different methods (FT-IR, UV-Vis, PL, SEM, and AFM analyses). The GM release profile displayed a controlled release manner over 48 h with a low initial burst release in the simulated wound media (PBS, pH 7.4). Antibacterial and in vitro cytotoxicity results showed a significant activity toward different gram-positive and negative bacterial strains (about 2.5 ± 0.1 cm inhibition zones) and a desired cytocompatibility against Human skin fibroblast (HFF-1) cells (over 80% cell viability), respectively. CONCLUSION: The obtained results recommend CQDs-crosslinked CS (CS/CQD) nanocomposite as a potent antimicrobial wound dressing.

Antisense Agents against Antibiotic-resistant Bacteria.

The dramatically increasing levels of antibiotic resistance are being seen worldwide and are a significant threat to public health. Antibiotic and drug resistance is seen in various bacterial species. Antibiotic resistance is associated with increased morbidity and mortality and increased treatment costs. Antisense-related technologies include oligonucleotides that interfere with gene transcription and expression; these oligonucleotides can help treat antibiotic-resistant bacteria. The important oligonucleotides include Peptide Nucleic Acids (PNAs), Phosphorodiamidate Morpholino Oligomers (PPMOs), and Locked Nucleic Acids (LNAs). Typically, the size of these structures (oligonucleotides) is 10 to 20 bases. PNAs, PPMOs, and LNAs are highlighted in this review as targets for genes that cause the gene to be destroyed and impede bacterial growth. These results open a new perspective for therapeutic intervention. Future studies need to examine different aspects of antisense agents, such as the safety, toxicity, and pharmacokinetic properties of antisense agents in clinical treatment.

Immunological Properties of Exotoxin A Toxoid - Detoxified Lipopolysaccharide - Gold Nanoparticles Conjugate Against Pseudomonas aeruginosa Infection.

BACKGROUND: Pseudomonas aeruginosa is an important opportunistic pathogen, especially in patients with compromised host defense. OBJECTIVE: To prepare the conjugate of detoxified lipopolysaccharide (D-LPS) and exotoxin A toxoid (T-ETA) from P. aeruginosa in gold nanoparticles (Au NPs) in a mice model. METHODS: LPS and ETA were purified from P. aeruginosa PAO1. D-LPS was conjugated withT-ETA via the amidation method. Au NPs were bound to D-LPS-T-ETA conjugate via electrostatic interaction. Mice were immunized with D-LPS, D-LPS-Au NPs, T-ETA, T-ETA-Au NPs, D-LPS-T-ETA, D-LPS-T-ETA-Au NPs, D-LPS-Au NPs+T-ETA-Au NPs, Au NPs, and phosphate-buffered saline (PBS), and specific IgG titers were determined by the ELISA and the whole-cell ELISA methods. Mice in the vaccinated and control groups were exposed to a 2×LD50 of P. aeruginosa and mortality rates were recorded for one week. RESULTS: The results showed that vaccination by D-LPS, D-LPS-Au NPs, T-ETA, T-ETA-Au NPs, D-LPS-T-ETA, D-LPS-T-ETA-Au NPs and D-LPS-Au NPs+T-ETA-Au NPs induced specific IgG. Mice received the D-LPS-T-ETA-Au NPs conjugate showed significant protection against bacterial challenge. CONCLUSION: These data indicate that D-LPS-T-ETA-Au NPs conjugate has a significant immunogenicity potential to be applied as a new vaccine against Pseudomonas infections.

Unsaturated fatty acids as a co-therapeutic agents in cancer treatment.

Chemotherapy is standard treatments for many malignancies. However, in most cases, this method is not able to induce apoptosis and in many cases, with cancer recurrence, leads to patient death. There are several procedure to control and suppress malignant cells, but among these methods, administration of É·-3 fatty acids and É·-6 fatty due to their destructive effects on cancer cells is more prominent. Many clinical studies have shown beneficial effects of É·-3 and É·-6 fatty acids in cardiovascular disorders, asthma, rheumatoid arthritis, osteoporosis and in most cancers such as colon, breast, prostate and other malignancies. Studies showed that polyunsaturated fatty acids (PUFAs) have a toxic effect on cancer cells. However, the exact mechanism of how É·- fatty acids affect cancer cells is still unknown. In this review alternative issues of malignancies co-treatments agents such as PUFAs have been studied. Also, the latest known PUFAs mechanisms on malignancies have been described.

Preparation and evaluation of the exotoxin A nano-gold conjugate as a vaccine candidate for Pseudomonas aeruginosa infections.

OBJECTIVES: Pseudomonas aeruginosa is an opportunistic pathogen that is an important cause of nosocomial infections. This bacterium produces various virulence factors, among which exotoxin A is significantly involved in mortality and morbidity. In this study, we evaluated the immunogenicity of native exotoxin A extracted from the P. aeruginosa and its conjugation with gold nanoparticles in the animal model. MATERIALS AND METHODS: Exotoxin A was first extracted and purified from the culture medium of P. aeruginosa PAO1 by selective precipitation and dialysis. The gold nanoparticles were prepared using the Turkevich method and conjugated to the prepared exotoxin A by electrostatic force. The size and conjugation were confirmed using electron microscopy and Fourier transform infrared spectrometry (FTIR), respectively. The immunogenicity of prepared ExoA-gold nanoparticles was investigated in the mice model. RESULTS: The results indicated that nano-gold particles can be conjugated to the native exotoxin A with high efficiency. Immunogenicity investigation demonstrated that antibody titers produced against native exotoxin A and its conjugate to nano-gold particles are significant in a mouse model (P<0.005). Moreover, significant protection against 2×LD50 P. aeruginosa infection was observed in animals immunized with nano-gold-exotoxin A as compared with control groups (P=0.00). CONCLUSION: Our study indicated that exotoxin A can be produced with acceptable purity in the laboratory, and conjugated to gold nanoparticles. Based on these results nano-gold-exotoxin A conjugate is highly immunogenic and can be considered a potential vaccine candidate for P. aeruginosa infections.

Effect of Lactobacillus delbrueckii Subsp. lactis PTCC1057 on Serum Glucose, Fetuin-A ,and Sestrin 3 Levels in Streptozotocin-Induced Diabetic Mice.

Intake of probiotic bacteria may improve or preserve insulin sensitivity. Fetuin-A and sestrin 3 have emerged as promising candidate biomarkers for crucial roles in insulin signaling pathway. Therefore, the effect of oral supplementation with the probiotic bacterium Lactobacillus delbrueckii subsp. lactis PTCC1057 on proteins involved in insulin signaling pathway was investigated in normal and streptozotocin (STZ)-induced diabetic mice. The 6-8-week-old female mice were divided into a non-diabetic control, diabetic control, and diabetic experimental and non-diabetic experimental groups (5 mice each group). Diabetic and non-diabetic experimental groups treated with 3 × 107 CFU mL-1 L. delbrueckii subsp. lactis PTCC1057 by gavage feeding approach daily for 28 days. Serum glucose, fetuin-A, and sestrin 3 levels were measured by standard methods. The result showed that oral administration of L. delbrueckii significantly decreased serum glucose in comparison to diabetic control group (P = 0.01). Serum fetuin-A level was higher in diabetic control group than non-diabetic group and oral administration of L. delbrueckii subsp. lactis PTCC1057 significantly decreased fetuin-A level in diabetic experimental group in comparison with non-diabetic groups (P = 0.001). Sestrin 3 level significantly was lower in diabetic control group than non-diabetic control group (P = 0.03) and it significantly increased in diabetic experimental group in comparison with diabetic control group after intervention of L. delbrueckii subsp. lactis PTCC1057 (P = 0.02). The results show that feeding the STZ-induced diabetic mice with L. delbrueckii subsp. lactis PTCC1057 terminated to decrease in fasting blood glucose and fetuin-A level and increase in serum sestrin 3 level. Therefore, the L. delbrueckii subsp. lactis PTCC1057 can be considered as excellent candidate for future studies on diabetes mellitus.

Biosynthesis of AgNPs onto the urea-based periodic mesoporous organosilica (AgxNPs/Ur-PMO) for antibacterial and cell viability assay.

Nano-size silver particles were stabilized on the inner surfaces of urea based periodic mesoporous organosilica (Ur-PMO). Aqueous extract of Euphorbia leaves as a sustainable and green reducing agent was applied for Ag-nanoparticles growth into the Ur-PMO channels. Physical and chemical properties of organosilica materials synthesized using various techniques such as FT-IR, small-angle XRD, PXRD, FESEM, TEM, SEM-EDX and atomic absorption spectrometry (AAS) were examined. Finally, the AgNPs/Ur-PMO were investigated on cell viability assay. An in vitro cytotoxicity test using MMT assay displayed that the designed material has good biocompatibility and could be a promising candidate for biomedical applications. The results also showed that the AgNPs/Ur-PMO compounds (especially, PMO; 1.27% AgNPs) had relatively good antibacterial and antibiofilm effects. It seems that the use of these compounds in hospital environments can reduce nosocomial infections as well as reduce antibiotic-resistant bacteria.

Poly (ε-Caprolactone)/Cellulose Nanofiber Blend Nanocomposites Containing ZrO2 Nanoparticles: A New Biocompatible Wound Dressing Bandage with Antimicrobial Activity.

Purpose: In the present study, the poly (ε-caprolactone)/cellulose nanofiber containing ZrO2 nanoparticles (PCL/CNF/ZrO2 ) nanocomposite was synthesized for wound dressing bandage with antimicrobial activity. Methods: PCL/CNF/ZrO2 nanocomposite was synthesized in three different zirconium dioxide amount (0.5, 1, 2%). Also the prepared nanocomposites were characterized by Infrared spectroscopy (FT-IR), X-ray diffraction (XRD), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). In addition, the morphology of the samples was observed by scanning electron microscopy (SEM). Results: Analysis of the XRD spectra showed a preserved structure for PCL semi-crystalline in nanocomposites and an increase in the concentrations of ZrO2 nanoparticles, the structure of nanocomposite was amorphous as well. The results of TGA, DTA, DSC showed thermal stability and strength properties for the nanocomposites which were more thermal stable and thermal integrate compared to PCL. The contact angles of the nanocomposites narrowed as the amount of ZrO2 in the structure increased. The evaluation of biological activities showed that the PCL/CNF/ZrO2 nanocomposite with various concentrations of ZrO2 nanoparticles exhibited moderate to good antimicrobial activity against all tested bacterial and fungal strains. Furthermore, cytocompatibility of the scaffolds was assessed by MTT assay and cell viability studies proved the non-toxic nature of the nanocomposites. Conclusion: The results show that the biodegradability of nanocomposite has advantages that can be used as wound dressing.

Thymol, cardamom and Lactobacillus plantarum nanoparticles as a functional candy with high protection against Streptococcus mutans and tooth decay.

Due to the increasing resistance of microorganisms against antibiotics, the use of natural bioactive substances for the prevention of pathogenic bacteria is considered in food products. In this study, thymol, cardamom essential oil, L. plantarum cell-free supernatant (ATCC 14917), and their nanoparticle candies prepared and inhibition activities against S. mutans (ATCC 25175), which is important in causing tooth decay, was investigated. Moisture content, pH, and sensory analyzes of candies measured. Also, SEM and FTIR of treated candy samples were performed. All examined bioactive substances and their nanoparticles showed an inhibitory effect against S. mutans with different minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The prepared candies had pH 5.5 represented a bactericidal effect against S. mutans. SEM and FTIR results approved the antibacterial effects of prepared candies. According to the results, all of the prepared candies significantly decreased S. mutans in saliva compared to the control candy and they are suitable agents for S. mutans growth-inhibiting. Also, cardamom essential oil candy showed the most general acceptance in a sensory analysis by panelists.

Chitosan biomaterials application in dentistry.

Chitosan is a natural biopolymer derived from deacetylation of chitin and it has been investigated with interdisciplinary approaches for multitude applications. Chitosan biomaterials possess unique properties such as biocompatibility, biodegradability, non-toxicity, muco-adhesion and a wide range of antibacterial and antifungal activity. Additionally, chitosan is the only cationic polysaccharide in nature and can be chemically modified to derivatives, based on the aim of function and application. The distinctive properties of chitosan and its derivatives have aroused interest in pharmaceutical industries and biomedical fields worldwide. This review discusses the crucial role of chitosan in production of bio-dental materials and accentuates its current profitable utilizations in oral drug delivery system, bone tissue engineering for treatment of periodontitis and dentin-pulp regeneration. Chemical modifications and incorporation of diverse bioactive molecules in order to improve the mechanical and biological characteristics of chitosan have also been discussed.

The critical role of Faecalibacterium prausnitzii in human health: An overview.

Faecalibacterium prausnitzii (F. prausnitzii) is one of the most abundant bacterial species in the colon of healthy human adults and representing more than 5% of the total bacterial population. Recently, it has been known as a major actor in human intestinal health and a biosensor. Changes in this species population richness and quantity have been observed in many illnesses and several investigations have reported that abundance of F. prausnitzii is reduced in different intestinal disorders. In the current review, we aim to consider literature from various library databases and electronic searches (Science Direct, PubMed, and Google Scholar) which were randomly collected and serve as an overview of different features of F. prausnitzii including metabolites, anti-inflammatory action, and correlation of dysbiosis of this bacterium with various complications in human.

How CRISPR-Cas System Could Be Used to Combat Antimicrobial Resistance.

Rapid emergence of antibiotic-resistant bacteria has made it harder for us to combat infectious diseases and to develop new antibiotics. The clustered regularly interspaced short palindromic repeats - CRISPR-associated (CRISPR-Cas) system, as a bacterial adaptive immune system, is recognized as one of the new strategies for controlling antibiotic-resistant strains. The programmable Cas nuclease of this system used against bacterial genomic sequences could be lethal or could help reduce resistance of bacteria to antibiotics. Therefore, this study aims to review using the CRISPR-Cas system to promote sensitizing bacteria to antibiotics. We envision that CRISPR-Cas approaches may open novel ways for the development of smart antibiotics, which could eliminate multidrug-resistant (MDR) pathogens and differentiate between beneficial and pathogenic microorganisms. These systems can be exploited to quantitatively and selectively eliminate individual bacterial strains based on a sequence-specific manner, creating opportunities in the treatment of MDR infections, the study of microbial consortia, and the control of industrial fermentation.

Oral spirochetes: Pathogenic mechanisms in periodontal disease.

Periodontitis is an infectious inflammatory disease resulting from infection of biofilm forming bacteria. Several bacterial factors regulate inflammatory response and cause to tissue damage and loss of connection between gingival and tooth. Since bacterial virulence factors and also host immune responses have role, understanding of periodontal disease is complex, in overall we can say that in this disease epithelium is deleted by bacteria. Oral spirochetes are related to periodontitis, among them, Treponema denticola, have been associated with periodontal diseases such as early-onset periodontitis, necrotizing ulcerative gingivitis, and acute pericoronitis. This review will analyse mechanisms of pathogenesis of spirochetes in periodontitis. Microorganisms cause destruction of gingival tissue by two mechanisms. In one, damage results from the direct action of bacterial enzymes and cytotoxic products of bacterial metabolism. In the other, only bacterial components have role, and tissue destruction is the inevitable side effect of a subverted and exaggerated host inflammatory response to plaque antigens.

Long non-coding RNA molecules in tuberculosis.

Tuberculosis (TB), a chronic disease caused by Mycobacterium tuberculosis, is one of the deadliest infectious diseases in the world. Despite significant advances in detection techniques and therapeutic approaches for tuberculosis, there is still no suitable solution for early screening and reducing the number of individuals affected and their effective treatment. Various cellular events can disrupt the development of TB. The basis of these events is dysregulating of genes expression patterns related with specific molecules. Long non-coding RNAs (lncRNAs) are molecules discovered to regulate the expression of protein-coding genes and participate in gene silencing, cell cycle regulation and cellular differentiation processes. Dysregulation of lncRNAs has been found to be associated with many diseases, including cancers and infectious diseases. Thus, the recognition of lncRNAs as novel molecular biomarkers and therapeutic targets for tuberculosis is promising. In the present review, we try to summarize the current findings of lncRNA expression patterns and its role in tuberculosis infection process.

Acinetobacter baumannii Efflux Pumps and Antibiotic Resistance.

Acinetobacter baumannii is a nosocomial pathogen and gram-negative coccobacillus that is responsible for opportunistic infections, pneumonia, and infections of the urinary tract, bloodstream, skin, and soft tissue. This bacterium poses a major public health problem due to inducing resistance to several drugs, isolates, multidrug treatment, and occasionally pan drugs. Drug resistance is not only a major concern caused by A. baumannii but also is considered as the main challenge in many other pathogens. Several factors such as the efflux pump are associated with antibiotic resistance, biofilm production, and genetic mutations. In this review, A. baumannii is introduced in then some of the practical works conducted on the existing efflux pump are reviewed. The importance of the efflux pump is considered in this paper in relation to the antibiotic resistance and mechanisms developed for the inhibition of these pumps as well.

AdeB efflux pump gene knockdown by mRNA mediated peptide nucleic acid in multidrug resistance Acinetobacter baumannii.

Multidrug-resistant Acinetobacter baumannii isolates cause critical problems in health-care environments. AdeABC is a resistance-nodulation-cell division (RND)-type multidrug efflux pump conferring resistance to clinically essential antibiotics in A. baumannii, such as ciprofloxacin. This study aimed to target adeB gene with antisense peptide nucleic acid (PNA) and investigate its effect on resistance to antibiotics. NCBI database was used to design appropriate PNA to target adeB gene, by connecting PNA to mRNA, the translation of mRNA can be prevented. Three clinical isolates and A. baumannii ATCC 17978 were treated with the designed PNA by electroporation and competence procedure. Minimum Inhibitory concentration (MIC) of ciprofloxacin, colistin, and tetracycline were determined by microbroth dilution method. In addition, the expression level of adeB gene was measured by quantitative real-time PCR (qRT-PCR). Isolates used in this study had mutations in gyrA and parC genes corresponding to resistance to ciprofloxacin. MIC of resistance to ciprofloxacin after treatment with PNA was reduced from 32 µg/ml to16 µg/ml in A. baumannii ATCC 17978 isolate. Susceptibility level of tetracycline, in the 2 clinical isolates was decreased from 64 µg/ml to 32 µg/ml and in the other isolate was reduced from 128 µg/ml to 64 µg/ml. The expression level of adeB gene was decreased in A. baumannii ATCC 17978 (P > 0.01) but not in clinical isolate (P = 0.107). Findings of the present study indicate overexpression of adeB efflux pump has extra effect on resistance to antibiotics in isolates with a defined mechanism of resistance. Antisense technology is a feasible technique to suppress the function of these genes, which may be further exploited to control multidrug-resistant isolates.

Peptide nucleic acid-mediated re-sensitization of colistin resistance Escherichia coli KP81 harboring mcr-1 plasmid.

Escherichia coli is a gram-negative bacterium and it causes a variety of diseases in humans. It causes a wide range of clinical infections in humans; urinary tract infections is the most prevalent infection caused by uropathogenic Escherichia coli. In recent years, the observation of antibiotic-resistant genes such as resistance to colistin, makes the Escherichia coli resistant to antibiotics like colistin (polymyxin E), because of that the use of new therapies like peptide nucleic acid (PNA) has attracted the consideration of scientists. The aim of this study is the assessment of the inhibitory role of PNA against mcr-1 gene and reduction of mcr-1 gene expression and MIC in colistin resistant E. coli by PNA. NCBI database was used to design PNA. Our study was carried out on E. coli KP81 bacteria containing the mcr-1 gene. Microbroth dilution (MIC) method was used to survey phenotypic sensitivity and determine the sensitivity of the bacteria to the colistin antibiotic. E. coli KP81 isolates were further investigated by polymerase chain reaction to assess the presence of mcr-1 genes and target genes were quantified by real-time PCR assay using specific primers. The MIC result after treatment with specific PNA showed that the resistance to colistin reduced about three fold and the resistance level dropped from 32 µg/ml to 4 µg/ml. The expression analysis of mcr-1 gene in E. coli KP81 isolate indicates the PNA, 95% reduced the expression of the mcr-1 gene. Our observations showed that by inhibiting the expression of mcr-1, sensitivity to colistin can be defeated. Using higher concentrations of PNA and an in vivo study can reveal more clinical application of this method.

Fabrication and characterization of a titanium dioxide (TiO2) nanoparticles reinforced bio-nanocomposite containing Miswak (Salvadora persica L.) extract - the antimicrobial, thermo-physical and barrier properties.

Objective: The microbial, physico-chemical and optical corruptions threaten a variety of foods and drugs and consequently the human biological safety and its accessible resources. The humanbeing's tendency towards bio-based materials and natural plant-extracts led to an increase in the usage of antimicrobial biocomposites based on medicinal herbs. Miswak (Salvadora persica L.) extract (SPE) has been proved effective for its antimicrobial and other biological activities. Therefore, in this study, titanium dioxide (TiO2) nanoparticles (TONP) and SPE were applied to fabricate antimicrobial carboxymethyl cellulose (Na-CMC) based bio-nanocomposites which would simultaneously promote some thermo-physical and barrier properties. Methods: CMC-neat film (C1), CMC/TONP-2% (C2) and CMC/TONP-2% with 150, 300 and 450 mg/mL SPE (SPE150, SPE30 and SPE450, respectively) were fabricated. The physical and mechanical properties; elemental mapping analysis (MAP), X-ray diffraction (XRD), scanning electron microscopy (SEM), thermal gravimetric analysis (TGA-DTG); fourier transform infrared (FTIR), energy-dispersive X-ray (EDX) and UV-vis spectroscopies were done to further validate the results. Results: Addition of TONP (2%) improved the blocking of UV light at 280 nm while SPE-containing nanocomposites completely blocked it. FTIR, XRD and SEM confirmed the formation of homogeneous films and high miscibility of applied materials. TONP led to an increase in Young's modulus (YM) and stress at break (SB) while SPE decreased them and enhanced the elongation to break (EB) (flexibility) of the active nanocomposites. Compared to CMC-film, the thermo-gravimetric analysis (TGA-DTG) showed a higher thermal stability for CMC/TONP and CMC/TONP/SPE nanocomposites. The EDX spectroscopy and elemental mapping analysis (MAP) proved the existence and well-distributedness of Na, K, Cl, S, Ti, F and N elements in SPE-activated nanocomposites. The pure SPE and SPE-activated nanocomposites showed a favorable antimicrobial activity against both gram-positive (Staphylococcus aureus) and negative (Escherichia coli) bacteria. Conclusion: The CMC-TiO2-SPE nanocomposites were homogeneously produced. Combination of TiO2 nanoparticles and dose-dependent SPE led to an improvement of thermal stability, and high potential in antimicrobial and UV-barrier properties. These results can generally highlight the role of the fabricated antimicrobial bio-nanocomposites as a based for different applications especially in food/drug packaging or coating.

Molecular mechanisms related to colistin resistance in Enterobacteriaceae.

Colistin is an effective antibiotic for treatment of most multidrug-resistant Gram-negative bacteria. It is used currently as a last-line drug for infections due to severe Gram-negative bacteria followed by an increase in resistance among Gram-negative bacteria. Colistin resistance is considered a serious problem, due to a lack of alternative antibiotics. Some bacteria, including Pseudomonas aeruginosa, Acinetobacter baumannii, Enterobacteriaceae members, such as Escherichia coli, Salmonella spp., and Klebsiella spp. have an acquired resistance against colistin. However, other bacteria, including Serratia spp., Proteus spp. and Burkholderia spp. are naturally resistant to this antibiotic. In addition, clinicians should be alert to the possibility of colistin resistance among multidrug-resistant bacteria and development through mutation or adaptation mechanisms. Rapidly emerging bacterial resistance has made it harder for us to rely completely on the discovery of new antibiotics; therefore, we need to have logical approaches to use old antibiotics, such as colistin. This review presents current knowledge about the different mechanisms of colistin resistance.