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2021 ◽  
Author(s):  
Roni F Rayes ◽  
Marnie G Wilson ◽  
Stephen D Gowing ◽  
France Bourdeau ◽  
Betty Giannias ◽  
...  

Background: Lung cancer is a leading cause of death partially due to high recurrence rates after surgical resection. Clinical data suggest that post-operative infections may increase the risk of recurrence. Our previous work indicated that increased adhesion of circulating tumors in the context of infection is partially responsible for this phenotype. However, cancer metastasis is a multi-step process, and it is likely that other events following tumor adhesion also play a role. Methods: In vivo intrasplenic injection of murine lung cancer cells into wild type (WT) and Toll-like receptor 4 knockout (TLR4-/-) mice followed by cecal-ligation and puncture (CLP) as a model of post-operative infection or sham surgery were used. H&E staining and immunohistochemistry analysis of Ki67+ cells in the livers of those mice were performed. In vitro proliferation assays were performed on human lung cancer cells using combinations of TLR blockade. Results: We found a 5-fold increase in hepatic metastases in WT CLP mice compared to WT sham mice. TLR4-/- CLP mice had a significant decreased tumor burden compared to WT CLP mice. This indicated an important mechanistic role for the TLR4-initiated host response to gram negative infection post-tumor cell adhesion. By analyzing the livers of those mice, we observed an increase in proliferation of tumor micrometastases in vivo in WT CLP mice as compared to WT sham mice. Here again, CLP TLR4 -/- mice had significantly fewer replicating micrometastases than CLP WT mice. Indeed, we found that direct stimulation of lung cancer cells with heat-inactivated E.Coli resulted in increased proliferation of tumor growth in vitro. These effects were partially abrogated by tumor TLR4 blockade; combined TLR2, 4 and 5 blockades led to a more prominent decrease. Conditioned media from bronchoalveolar epithelial cells treated with lipopolysaccharide lead to increased lung cancer proliferation; these changes were reversed with TLR blockade, indicating that the host response to infection is TLR mediated. Conclusions: Overall, these results imply a more complex mechanistic role of post-operative infection in metastasis. From a clinical standpoint, this evidence strengthens the case for the use of TLR blockade as a potential therapeutic target in the prevention of metastasis.


2021 ◽  
Vol 15 (1) ◽  
pp. 92-97
Author(s):  
Valentina Paci ◽  
Pierina Visciano ◽  
Ivanka Krasteva ◽  
Tiziana Di Febo ◽  
Fabrizia Perletta ◽  
...  

Background: The diagnosis of brucellosis by serological tests is based on antigen suspensions derived from smooth lipopolysaccharide extracts, which can give false positive results linked to cross-reactivity with other Gram-negative microorganisms, especially Yersinia enterocolitica O:9 and Escherichia coli O157:H7. Objective: The objective of the present study was the characterization by proteomic analysis of specific immunogenic proteins not associated with smooth lipopolysaccharide to improve the diagnostic tests used in the ovine brucellosis eradication programs. Methods: The serum from a sheep positive to Brucella melitensis was treated to eliminate all antibodies against such lipopolysaccharide and highlight the reaction towards the immunoreactive proteins in Western Blotting. Results: The immunoreactive bands were identified by nLC-MS/MS and through bioinformatic tools, it was possible to select 12 potential candidates as protein antigens specific for Brucella melitensis. Conclusion: The detection of new antigens not subjected to cross-reactivity with other Gram-negative microorganisms can offer an additional tool for the serological diagnosis of such disease.


2021 ◽  
Vol 8 ◽  
Author(s):  
Nazanin Mansouri Shirazi ◽  
Niloofar Eslahi ◽  
Adeleh Gholipour-Kanani

Keratin protein has been applied for biomedical applications due to its biocompatibility, biodegradability, mechanical resistance, and bioavailability. Tragacanth gum (TG) as a polysaccharide-based biopolymer has wound healing and antimicrobial properties. In this study, keratin was extracted from protein-based chicken feather by using reduction hydrolysis (sodium sulfide), and nanogels of keratin and TG composites at different ratios were produced by using the chemical cross-linking method. Then, cinnamon (5 and 10%) as an antibacterial herbal extract was added to the nanogels and coated on cotton fabric. The morphology and size of the composite nanogels, chemical structure, biological, and antibacterial properties were evaluated. According to DLS results, TGK2:1 (ratio of TG to keratin = 2:1) had the minimum size (80 nm) and PDI (0.1), and therefore, this sample was chosen as the optimum one. FESEM and TEM images showed the semi-spherical shape of the produced nanogels. FTIR spectra revealed the possible hydrogen bonding between the components, and the formation of disulfide bonds after the addition of hydrogen peroxide was confirmed by XPS. After loading cinnamon into the nanogels, an increase in size was observed from 80 nm for free-nanogel to 85 and 105 nm for 5 and 10% extract-loaded nanogels, respectively. Besides, more cinnamon was released from the treated fabrics by increasing time and cinnamon concentration. The antibacterial test exhibited good antibacterial properties against both Gram-positive and Gram-negative bacteria. Finally, MTT assay approved the biocompatibility of the produced nanogels for potential use in medical textiles.


2021 ◽  
Vol 218 (11) ◽  
Author(s):  
Annika Hausmann ◽  
Boas Felmy ◽  
Leo Kunz ◽  
Sanne Kroon ◽  
Dorothée Lisa Berthold ◽  
...  

Intestinal epithelial cell (IEC) NF-κB signaling regulates the balance between mucosal homeostasis and inflammation. It is not fully understood which signals tune this balance and how bacterial exposure elicits the process. Pure LPS induces epithelial NF-κB activation in vivo. However, we found that in mice, IECs do not respond directly to LPS. Instead, tissue-resident lamina propria intercrypt macrophages sense LPS via TLR4 and rapidly secrete TNF to elicit epithelial NF-κB signaling in their immediate neighborhood. This response pattern is relevant also during oral enteropathogen infection. The macrophage–TNF–IEC axis avoids responses to luminal microbiota LPS but enables crypt- or tissue-scale epithelial NF-κB responses in proportion to the microbial threat. Thereby, intercrypt macrophages fulfill important sentinel functions as first responders to Gram-negative microbes breaching the epithelial barrier. The tunability of this crypt response allows the induction of defense mechanisms at an appropriate scale according to the localization and intensity of microbial triggers.


2021 ◽  
Vol 12 ◽  
Author(s):  
Carla L. Schwan ◽  
Sara Lomonaco ◽  
Leonardo M. Bastos ◽  
Peter W. Cook ◽  
Joshua Maher ◽  
...  

Non-typhoidal Salmonella enterica is a pathogen of global importance, particularly in low and middle-income countries (LMICs). The presence of antimicrobial resistant (AMR) strains in market environments poses a serious health threat to consumers. In this study we identified and characterized the genotypic and phenotypic AMR profiles of 81 environmental S. enterica strains isolated from samples from informal markets in Cambodia in 2018–2019. AMR genotypes were retrieved from the NCBI Pathogen Detection website (https://www.ncbi.nlm.nih.gov/pathogens/) and using ResFinder (https://cge.cbs.dtu.dk/services/) Salmonella pathogenicity islands (SPIs) were identified with SPIFinder (https://cge.cbs.dtu.dk/services/). Susceptibility testing was performed by broth microdilution according to the Clinical and Laboratory Standards Institute (CLSI) standard guidelines M100-S22 using the National Antimicrobial Resistance Monitoring System (NARMS) Sensititre Gram Negative plate. A total of 17 unique AMR genes were detected in 53% (43/81) of the isolates, including those encoding tetracycline, beta-lactam, sulfonamide, quinolone, aminoglycoside, phenicol, and trimethoprim resistance. A total of 10 SPIs (SPI-1, 3–5, 8, 9, 12–14, and centisome 63 [C63PI]) were detected in 59 isolates. C63PI, an iron transport system in SPI-1, was observed in 56% of the isolates (n = 46). SPI-1, SPI-4, and SPI-9 were present in 13, 2, and 5% of the isolates, respectively. The most common phenotypic resistances were observed to tetracycline (47%; n = 38), ampicillin (37%; n = 30), streptomycin (20%; n = 16), chloramphenicol (17%; n = 14), and trimethoprim-sulfamethoxazole (16%; n = 13). This study contributes to understanding the AMR genes present in S. enterica isolates from informal markets in Cambodia, as well as support domestic epidemiological investigations of multidrug resistance (MDR) profiles.


Author(s):  
Eliza Depoorter ◽  
Tom Coenye ◽  
Peter Vandamme

Burkholderia cepacia complex strain R-12632 produces ditropolonyl sulfide, an unusual sulfur-containing tropone, via a yet unknown biosynthetic pathway. Ditropolonyl sulfide purified from a culture of strain R-12632 inhibits the growth of various Gram-positive and Gram-negative multidrug resistant bacteria, with minimum inhibitory concentration (MIC) values as low as 16 μg/ml. In the present study we used a transposon mutagenesis approach combined with metabolite analyses to identify the genetic basis for antibacterial activity of strain R-12632 against Gram-negative bacterial pathogens. Fifteen of the 8304 transposon mutants investigated completely lost antibacterial activity against Klebsiella pneumoniae LMG 2095. In these loss-of-activity mutants, nine genes were interrupted. Four of those genes were involved in assimilatory sulfate reduction, two in phenylacetic acid (PAA) catabolism and one in glutathione metabolism. Via semipreparative fractionation and metabolite identification, it was confirmed that inactivation of the PAA degradation pathway or glutathione metabolism led to loss of ditropolonyl sulfide production. Based on earlier studies on the biosynthesis of tropolone compounds, the requirement for a functional PAA catabolic pathway for antibacterial activity in strain R-12632 indicated that this pathway likely provides the tropolone backbone for ditropolonyl sulfide. Loss of activity observed in mutants defective in assimilatory sulfate reduction and glutathione biosynthesis suggested that cysteine and glutathione are potential sources of the sulfur atom linking the two tropolone moieties. The demonstrated antibacterial activity of the unusual antibacterial compound ditropolonyl sulfide warrants further studies into its biosynthesis and biological role. Importance Burkholderia bacteria are historically known for their biocontrol properties and have been proposed as a promising and underexplored source of bioactive specialized metabolites. Burkholderia cepacia complex strain R-12632 inhibits various Gram-positive and Gram-negative resistant pathogens and produces numerous specialized metabolites, among which ditropolonyl sulfide. This unusual antimicrobial has been poorly studied and its biosynthetic pathway remained unknown. In the present study, we performed transposon mutagenesis of strain R-12632 and performed genome and metabolite analyses of loss-of-activity mutants to study the genetic basis for antibacterial activity. Our results indicate that the phenylacetic acid catabolism, assimilatory sulfate reduction and glutathione metabolism are necessary for ditropolonyl sulfide production. These findings contribute to understanding the biosynthesis and biological role of this unusual antimicrobial.


2021 ◽  
Author(s):  
Hani Abdel Hafez Naseef shtaya ◽  
Ula Mohammad ◽  
Nimeh Al-Shami ◽  
Yousef Sahoury ◽  
Abdallah D Abukhalil ◽  
...  

Diagnosis of co-infections with multiple pathogens among hospitalized COVID-19 patients can be jointly challenging and very essential for appropriate treatment, shortening hospital stay and preventing antimicrobial resistance. This study proposes to investigate the burden of bacterial and fungal co-infections outcomes on COVID-19 patients. It is a single centre cross-sectional study of hospitalized COVID-19 patients at Beit-Jala hospital in Palestine. The study included 321 hospitalized patients admitted to the ICU between June 2020 and March 2021 aged ≥20 years, with a confirmed diagnosis of COVID-19 via RT-PCR conducted on a nasopharyngeal swab. The patient's information was gathered using graded data forms from electronic medical reports. The diagnosis of bacterial and fungal infection was proved through the patient`s clinical presentation and positive blood or sputum culture results. All cases had received empirical antimicrobial therapy before the ICU admission, and different regimens during the ICU stay. The rate of bacterial co-infection was 51.1%, mainly from gram-negative isolates (Enterobacter species and K.pneumoniae). The rate of fungal co-infection caused by A.fumigatus was 48.9%, and the mortality rate was 8.1%. However, it is unclear if it had been attributed to SARS-CoV-2 or coincidental.


Genes ◽  
2021 ◽  
Vol 12 (9) ◽  
pp. 1421
Author(s):  
Yuqiang Zhao ◽  
Yanxin Wang ◽  
Chengyao Xia ◽  
Xu Li ◽  
Xianfeng Ye ◽  
...  

Corallococcus sp. strain EGB is a Gram-negative myxobacteria isolated from saline soil, and has considerable potential for the biocontrol of phytopathogenic fungi. However, the detailed mechanisms related to development and predatory behavior are unclear. To obtain a comprehensive overview of genetic features, the genome of strain EGB was sequenced, annotated, and compared with 10 other Corallococcus species. The strain EGB genome was assembled as a single circular chromosome of 9.4 Mb with 7916 coding genes. Phylogenomics analysis showed that strain EGB was most closely related to Corallococcus interemptor AB047A, and it was inferred to be a novel species within the Corallococcus genus. Comparative genomic analysis revealed that the pan-genome of Corallococcus genus was large and open. Only a small proportion of genes were specific to strain EGB, and most of them were annotated as hypothetical proteins. Subsequent analyses showed that strain EGB produced abundant extracellular enzymes such as chitinases and β-(1,3)-glucanases, and proteases to degrade the cell-wall components of phytopathogenic fungi. In addition, 35 biosynthetic gene clusters potentially coding for antimicrobial compounds were identified in the strain EGB, and the majority of them were present in the dispensable pan-genome with unexplored metabolites. Other genes related to secretion and regulation were also explored for strain EGB. This study opens new perspectives in the greater understanding of the predatory behavior of strain EGB, and facilitates a potential application in the biocontrol of fungal plant diseases in the future.


Polymers ◽  
2021 ◽  
Vol 13 (18) ◽  
pp. 3116
Author(s):  
Thien Bui-Thuan Do ◽  
Tien Ngoc-Thuy Nguyen ◽  
Minh Hieu Ho ◽  
Nghi Thi-Phuong Nguyen ◽  
Thai Minh Do ◽  
...  

(1) Background: Wounds with damages to the subcutaneous are difficult to regenerate because of the tissue damages and complications such as bacterial infection. (2) Methods: In this study, we created burn wounds on pigs and investigated the efficacy of three biomaterials: polycaprolactone-gelatin-silver membrane (PCLGelAg) and two commercial burn dressings, Aquacel® Ag and UrgoTulTM silver sulfadiazine. In vitro long-term antibacterial property and in vivo wound healing performance were investigated. Agar diffusion assays were employed to evaluate bacterial inhibition at different time intervals. Minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and time-kill assays were used to compare antibacterial strength among samples. Second-degree burn wounds in the pig model were designed to evaluate the efficiency of all dressings in supporting the wound healing process. (3) Results: The results showed that PCLGelAg membrane was the most effective in killing both Gram-positive and Gram-negative bacteria bacteria with the lowest MBC value. All three dressings (PCLGelAg, Aquacel, and UrgoTul) exhibited bactericidal effect during the first 24 h, supported wound healing as well as prevented infection and inflammation. (4) Conclusions: The results suggest that the PCLGelAg membrane is a practical solution for the treatment of severe burn injury and other infection-related skin complications.


2021 ◽  
Vol 2021 ◽  
pp. 1-7
Author(s):  
Dongkai Sun ◽  
Peishan Cong ◽  
Fengju Guan ◽  
Shuai Liu ◽  
Lijiang Sun ◽  
...  

Objective. We sought to analyze the distribution and antibiotic sensitivity of pathogens in hospitalized patients and to provide a scientific reference for the rational application of antibiotics. Methods. From January 2014 to December 2018, urine cultures from patients in our hospital were collected and analyzed retrospectively for the presence, distribution, and drug sensitivity of pathogens. Results. A total of 42,854 midstream urine cultures were collected from which 11,891 (27.75%) pathogens were isolated, including 8101 (68.13%) strains of gram-negative bacteria, 2580 (21.69%) strains of gram-positive bacteria, and 1210 (10.18%) strains of fungi. Escherichia coli and Enterococci were the most common species of gram-negative and gram-positive bacteria, respectively. Drug sensitivity varied among different pathogens. Clear drug resistance was observed in bacteria, while fungus exhibited relatively lower resistance. Conclusion. Pathogens responsible for urinary tract infections in hospitalized patients are diversiform and display resistance to some antibiotics. Drug resistance monitoring should be enhanced to optimize antimicrobial therapy.


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