Potential antibacterial activity and healing effect of topical administration of bone marrow and adipose mesenchymal stem cells encapsulated in collagen-fibrin hydrogel scaffold on full-thickness burn wound infection caused by Pseudomonas aeruginosa.

Burns injuries are prone to hospital-acquired infections, and Pseudomonas aeruginosa is one of the most common causes of mortality and morbidity in patients with burn injuries. Thus, this study aimed to analyze the effects of topical treatment with bone marrow (BM-MSC) and adipose mesenchymal stem cells (AD-MSC) encapsulated in collagen and fibrin scaffolds in a Balb/c model of burn wound infection. Extraction of stem cells from adipose and bone marrow tissue of rats was performed and cells were characterized using standard methods. Then, collagen, fibrin and collagen-fibrin scaffolds were constructed and the extracted cells were encapsulated in all three scaffolds. Then, 3rd degree burn was induced in mice and 1.5 × 108 (CFU/ml) of P. aeruginosa was introduced to the burn wound. Subsequently, after 24 h of inducing wound infection, encapsulated MSCs were introduced as dressings to burn wound infection and microbial load as well as rate of wound infection healing was measured. The results of this study showed that the use of BM-MSC and AD-MSC encapsulated in collagen-fibrin scaffold reduced the bacteria load down to 54 and 21 CFU/gr, respectively (P < 0.05). Moreover, BM-MSC and AD-MSC encapsulated in collagen-fibrin showed 80% and 75% wound healing, respectively (P < 0.05). Also, we found no significant between cell origin and healing. Encapsulation of MSCs into collagen-fibrin scaffolds could be effective not only against P. aeruginosa infection, but also healing and regeneration of burn wound.

Evaluation of Genetic Content of the CRISPR Locus in Listeria monocytogenes Isolated From Clinical, Food, Seafood and Animal Samples in Iran.

CRISPR arrays, which are organized to fight against non-self DNA elements, have shown sequence diversity that could be useful in evolution and typing studies. In this study, 55 samples of L. monocytogenes isolated from different sources were evaluated for CRISPR sequence polymorphism. The CRISPR loci were identified using CRISPR databases. A single PCR assay was designed to amplify the target CRISPRs using an appropriate universal primer. Sequencing results were analyzed using CRISPR databases and BLASTn, and the CRISPR locus was present in all the strains. Three hundred repeats including 55 terminal repeats were identified. Four types of consensuses direct repeat (DR) with different lengths and sequences were characterized. Sixty repeat variants were observed which possessed different polymorphisms. Two hundred and fifty spacers were identified from which 35 consensus sequences were determined, indicating the high polymorphism of the CRISPR spacers. The identified spacers showed similarities to listeria phage sequences, other bacterial phage sequences, plasmid sequences and bacterial sequences. In order to control the bacterial outbreaks, a robust and precise system of subtyping is required. High levels of polymorphism in the CRISPR loci in this study might be related to the origin and time of the samples' isolation. However, it is essential to assess, on a case-by-case basis, the characteristics of any given CRISPR locus before its use as an epidemiological marker. In conclusion, the results of this study showed that the use of sequence content of CRISPR area could provide new and valuable information on the evolution and typing of the L. monocytogenes bacterium.

Bacteriocins: Properties and potential use as antimicrobials.

A variety of bacteriocins originate from lactic acid bacteria, which have recently been modified by scientists. Many strains of lactic acid bacteria related to food groups could produce bacteriocins or antibacterial proteins highly effective against foodborne pathogens such as Staphylococcus aureus, Pseudomonas fluorescens, P. aeruginosa, Salmonella typhi, Shigella flexneri, Listeria monocytogenes, Escherichia coli O157:H7, and Clostridium botulinum. A wide range of bacteria belonging primarily to the genera Bifidobacterium and Lactobacillus have been characterized with different health-promoting attributes. Extensive studies and in-depth understanding of these antimicrobials mechanisms of action could enable scientists to determine their production in specific probiotic lactic acid bacteria, as they are potentially crucial for the final preservation of functional foods or for medicinal applications. In this review study, the structure, classification, mode of operation, safety, and antibacterial properties of bacteriocins as well as their effect on foodborne pathogens and antibiotic-resistant bacteria were extensively studied.

Transcriptome analysis of biofilm formation under aerobic and microaerobic conditions in clinical isolates of Campylobacter spp.

It has been well documented that Campylobacter is the leading cause of foodborne infections and bacterial enteritis in high-income countries. The gastrointestinal tract of most warm-blooded animals, such as mammals and poultry, is prone to this pathogen. Infections caused by this bacterium in humans have usually been associated with the consumption of contaminated poultry meat. The important point about Campylobacter is that this bacterium has adapted to harsh environmental conditions along the food chain (poultry digestive tract to the consumer's plate) and developed an adapted mechanism to those conditions. This study aimed to compare the ability of Campylobacter jejuni and Campylobacter coli strains to form biofilms under aerobic and microaerobic conditions. The presence and expression of flab, FliS, DnaK, luxs, CsrA, Cj0688, and cosR genes involved in biofilm formation were investigated. Finally, the correlation between the biofilm forming ability of Campylobacter isolates and the presence/expression of selected genes has been explored. A significant correlation was observed between the presence and expression of some genes and the degree of biofilm formation in C. jejuni and C. coli isolates. A strong biofilm production was detected in strains harboring all selected genes with greater expression levels. The ability of C. jejuni and C. coli strains in biofilm formation is associated with the coordinated function and convergent expression of the selected genes. Seemingly, stress response- and motility-related genes have the most involvement in biofilm formation of C. jejuni and C. coli strains, while other genes have an accessory role in this phenomenon.

Microbiological Detoxification of Mycotoxins: Focus on Mechanisms and Advances.

Some fungal species of the genera Aspergillus, Penicillium, and Fusarium secretes toxic metabolites known as mycotoxins, have become a global concern that is toxic to different species of animals and humans. Biological mycotoxins detoxification has been studied by researchers around the world as a new strategy for mycotoxin removal. Bacteria, fungi, yeast, molds, and protozoa are the main living organisms appropriate for the mycotoxin detoxification. Enzymatic and degradation sorptions are the main mechanisms involved in microbiological detoxification of mycotoxins. Regardless of the method used, proper management tools that consist of before-harvest prevention and after-harvest detoxification are required. Here, in this review, we focus on the microbiological detoxification and mechanisms involved in the decontamination of mycotoxins.

A genomic concept in cellular interaction of clinical Campylobacter spp. with human epithelial colorectal adenocarcinoma cells.

This study aimed to realize the genomic concept of cellular interaction of clinical Campylobacter spp. with human epithelial colorectal adenocarcinoma cells. It was indicated that the mean adherence and invasion rate of C.jejuni isolates was significantly higher than C.coli and the highest adhesion rate among the C.jejuni and C.coli belonged to strains harboring 4 (flaA, cadF, peb1A, and flpA) and 3 (flaA, cadF, and peb1A) adherence genes, respectively, which indicates that the adhesion potential of C.coli and C.jejuni strains is associated with the coordinate function and cumulative effect of selected virulence-associated genes. The highest invasion rate in C.jejuni (10.3%) and C.coli (8.4%) isolates belonged to strains which concomitantly contained 3 (ciaB, iamA, and tlp1) and 2 (ciaB and iamA) invasion-associated genes which emphasizes on the cooperative roles of these genes in C.jejuni and C.coli invasion to Caco-2 cells. The toxicity of C.jejuni for Caco-2 cells was proved higher than that of C.coli. There was a positive correlation between adherence, invasion and toxicity of both C.jejuni and C.coli isolates. Moreover, the expression levels of CDT-producing genes in C.jejuni strains was significantly higher than that of C.coli. The average cytotoxicity of the strains with all three CDT-encoding genes (cdtA, cdtB and cdtC) was statistically higher than those lacking one or more CDT subunits. A crucial contribution of CdtB to the cytotoxicity of Campylobacter strains was detected. Following the treatment of epithelial cells with C.jejuni or C.coli, IL-8 and TNF-α were significantly increased compared to untreated Caco-2 cells, and the highest IL-8 expression was observed in both C.jejuni and C.coli expressing all CDTs (cdtA, cdtB, and cdtC). We, for the first time, indicated the major contribution of TLR2 and TLR4 in campylobacter initiation of pathogenesis, while increased invasiveness and cytotoxicity was significantly associated with the increased expression of TLR4 in C.jejuni isolates.

Targeting Listeria monocytogenes consensus sequence of internalin genes using an antisense molecule.

As an intracellular pathogen, Listeria monocytogenes can enter host cells where it can replicate and escape detection and eradication by the host immune response making the clearance of infection very challenging. Furthermore, with the advent of antimicrobial resistance, the need for alternative targets is inevitable. Internalin proteins are crucial to this bacterium as they contribute to bacterial entry to the systemic circulation. In this study, we targeted a highly conserved region of these proteins by an antisense sequence that was covalently conjugated to the cell penetrating peptides (CPP) to overcome the challenging delivery barriers. Then, we evaluated the efficiency of this construct in vitro. We also assessed the antigenicity, cytotoxicity, and probability of apoptosis induction by this construct. The studied CPP-PNA inhibited bacterial growth and suppressed the mRNA expression of internalins in a dose-dependent manner. In addition, at all studied concentrations, CPP-PNA significantly reduced the invasion rate of L. monocytogenes in the examined cell lines. Moreover, different concentrations of CPP-PNA did not have a significant antigenic, cytotoxic, and apoptotic properties compared to the control. These results suggest the effectiveness of CPP-antisense in targeting the mRNAs of internalins for various research, therapeutic and preventive purposes. However, additional research is required to evaluate the potency, safety, and pharmacokinetics of this compound for the prevention and treatment of listeriosis.

Time-variable expression levels of mazF, atlE, sdrH, and bap genes during biofilm formation in Staphylococcus epidermidis.

Staphylococcus epidermidis is an opportunistic pathogen causing infections related to the usage of implants and medical devices. Pathogenicity of this microorganism is mainly linked to its capability to form biofilm structures. Biofilm formation vastly depends on several factors including different proteins. We studied the expression levels of three proteins including SdrH, Bap, AtlE, and MazF at different time intervals during the course of biofilm formation. In this study, a catheter-derived S. epidermidis isolate with strong ability of biofilm formation was selected. PCR assay was used to detect sdrH, bap, atlE, and mazF genes in this isolate. Real-time PCR was used to determine the expression levels of these genes after 4, 8, and 20 h during the course of biofilm formation. The studied genes showed different expression levels at different time intervals during biofilm formation by real-time PCR method. Expression levels of atlE and sdrH genes were the highest at 4 h, whereas bap gene showed the highest expression level at 8 h during the course of biofilm formation. In addition, the expression level of mazF gene peaked at 4 h and then progressively decreased at 8 and 20 h. Our results suggest the importance of AtlE, SdrH, and MazF proteins in the establishment and development of the biofilm structure. In addition, our results showed the important role of protein Bap in the accumulation of biofilm structure. Future studies are required to understand the exact role of MazF in the process of biofilm formation.

Therapeutic bacteria to combat cancer; current advances, challenges, and opportunities.

Successful treatment of cancer remains a challenge, due to the unique pathophysiology of solid tumors, and the predictable emergence of resistance. Traditional methods for cancer therapy including radiotherapy, chemotherapy, and immunotherapy all have their own limitations. A novel approach is bacteriotherapy, either used alone, or in combination with conventional methods, has shown a positive effect on regression of tumors and inhibition of metastasis. Bacteria-assisted tumor-targeted therapy used as therapeutic/gene/drug delivery vehicles has great promise in the treatment of tumors. The use of bacteria only, or in combination with conventional methods was found to be effective in some experimental models of cancer (tumor regression and increased survival rate). In this article, we reviewed the major advantages, challenges, and prospective directions for combinations of bacteria with conventional methods for tumor therapy.

Evaluation of Fosfomycin Activity Against Extended Spectrum Beta Lactamase (ESBL) Producing Enterobacteriaceae Isolated from Three Centers of Tehran, Iran.

BACKGROUND: Rising rates of antimicrobial resistance among Enterobacteriaceae limit the use of reliably active forms of available drugs. The aim of this study was to investigate the prevalence of fosfomycin (US6794490B2) resistance gene among ESBL producing isolates in Iran. METHOD: We tested 355 isolates of Enterobacteriacea collected from various clinical samples including urine, wounds, blood and other sources during June 2016 to July 2017. Antibiotic sensitivity and Extended Spectrum Beta Lactamase (ESBL) production were tested using agar dilution method according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. ESBL genes (blaTEM, bla SHV,bla CTX-M), plasmid-encoded fosfomycin resistance genes (fosA, fosB, fosA3 and fosC2) and chromosomal mutations (murA, glpT, uhpT) were detected by Polymerase Chain Reaction (PCR). RESULTS: In this study, 151 of the 355 isolates were ESBL-positive. blaCTX-M (77%) was the most common gene followed by blaSHV (70%) and blaTEM (58%), either alone or in combination. Eighty nine percent (132/151) of the ESBL-positive isolates were MDR. Antimicrobial susceptibility rates were higher for fosfomycin (92.8%) and imipenem (35.5%) among ESBL-positive isolates. None of the ESBL- positive isolates harbored any mutations or plasmid-mediated fosfomycin resistance determinants. CONCLUSION: In conclusion, fosfomycin showed good antimicrobial activity against multidrug resistance ESBL- positive Enterobacteriaceae.