Bacterial Penetration Research Articles

A surfactant-based dressing can reduce the appearance of Pseudomonas aeruginosa pigments and uncover the dermal extracellular matrix in an ex vivo porcine skin wound model.

From previous studies, we have shown that viable colony forming units of bacteria and bacterial biofilms are reduced after sequential treatment with a surfactant-based dressing. Here, we sought to test the impact on visible bacterial pigments and the ultrastructural impact following the sequential treatment of the same surfactant-based dressing. Mature Pseudomonas aeruginosa biofilms were grown on ex vivo porcine skin explants, and an imaging-based analysis was used to compare the skin with and without a concentrated surfactant. In explants naturally tinted by bacterial chromophores, wiping alone had no effect, while the use of a surfactant-based dressing reduced coloration. Similarly, daily wiping led to increased immunohistochemical staining for P. aeruginosa antigens, but not in the surfactant group. Confocal immunofluorescent imaging revealed limited bacterial penetration and coating of the dermis and loose pieces of sloughing material. Ultrastructural analysis confirmed that the biofilms were masking the extracellular matrix (ECM), but the surfactant could remove them, re-exposing the ECM. The masking of the ECM may provide another non-inflammatory explanation for delayed healing, as the ECM is no longer accessible for wound cell locomotion. The use of a poloxamer-based surfactant appears to be an effective way to remove bacterial chromophores and the biofilm coating the ECM fibres.

Oxygen‐Free Polycationic Photosensitizers for Treatment of Periodontal Inflammation

AbstractPeriodontal tissue infection is a chronic destructive inflammatory disease caused by bacteria including anaerobes and facultative anaerobes. Traditional mechanical debridement and antibiotic treatments are not effective at removing biofilm from the root surface, resulting in recurrent disease. Antimicrobial photodynamic therapy (aPDT) is a promising bactericidal method, but it is limited by oxygen content. Here, an oxygen‐free cationic polymer (HQRB‐SS‐Dex) containing the photosensitizer rose bengal (RB) and dextran is proposed. It releases both 1O2 and HO•, and it exhibits strong antibacterial efficiency against periodontal pathogenic bacteria under irradiation. The positively charged quaternary ammonium polymer and dextran of HQRB‐SS‐Dex facilitate adherence to the bacterial surface and penetration of the bacterial biofilm to permit efficient use of reactive oxygen species. Disulfide bonds make the complex biodegradable, which improves biosafety. This work presents a unique branched cationic polymer for oxygen‐free aPDT, with potential application to the treatment of periodontal infection.

Gut Bacterial Community Determines the Therapeutic Effect of Ginsenoside on Canine Inflammatory Bowel Disease by Modulating the Colonic Mucosal Barrier.

Inflammatory bowel disease (IBD) comprises systemic inflammatory conditions primarily affecting the gastrointestinal tract, including Crohn's disease and ulcerative colitis. This research aims to analyze the clinical symptoms and pathogenesis of a Dextran sodium sulfate (DSS)-induced canine IBD model and evaluate the restorative effect of ginsenoside from a pathogenesis perspective. We established the DSS-induced canine IBD model and studied the pathological mechanisms. Additionally, we examined the therapeutic effect of ginsenosides by assessing the Canine Inflammatory Bowel Disease Activity Index (CIBDAI), C-reactive protein (CRP) levels, colonic tissue morphology, protein expression, and mucosal bacterial community analysis. Our findings revealed a total ginsenoside content of 22.7% in the ginsenoside extract. Animal experiments demonstrated that dogs with IBD exhibited decreased mental state, significantly increased CIBDAI and CRP levels, disrupted colonic epithelial tissue structure, decreased expression of mucin, tight junctions, and adherens junctions, as well as reduced diversity of the colonic mucosal bacterial community. Furthermore, correlation analysis highlighted a total of 38 bacterial strains correlated with physiological indices. Significantly, ginsenoside treatment could improve these symptoms and reverse the relative abundance of some bacterial communities. In conclusion, alterations in the properties of the colonic mucus layer or the reduction in MUC2, its core component, in dogs with IBD can lead to bacterial penetration of the mucus layer and subsequent contact with intestinal epithelial cells, resulting in inflammation. Remarkably, ginsenoside intervention showcased the capacity to positively influence the relative abundance of bacteria and impact the colonic mucus layer properties, thereby offering promising prospects for IBD management and recovery.

Photodynamic inactivation of bacteria: Why it is not enough to excite a photosensitizer

BackgroundThe development of multidrug resistance (MDR) in infectious agents is one of the most serious global problems facing humanity. Antimicrobial photodynamic therapy (APDT) shows encouraging results in the fight against MDR pathogens, including those in biofilms. MethodsPhotosensitizers (PS), monocationic methylene blue, polycationic and polyanionic derivatives of phthalocyanines, electroneutral and polycationic derivatives of bacteriochlorin were used to study photodynamic inactivation of Gram-positive and Gram-negative planktonic bacteria and biofilms under LED irradiation. Zeta potential measurements, confocal fluorescence imaging, and coarse-grained modeling were used to evaluate the interactions of PS with bacteria. PS aggregation and photobleaching were studied using absorption and fluorescence spectroscopy. ResultsThe main approaches to ensure high efficiency of bacteria photosensitization are analyzed. ConclusionsPS must maintain a delicate balance between binding to exocellular and external structures of bacterial cells and penetration through the cell wall so as not to get stuck on the way to photooxidation-sensitive structures of the bacterial cell.

Bacterial contamination of medical face mask wearing duration and the optimal wearing time.

Bacterial contamination is a critical parameter for how long a medical mask will be worn. In this study, we used the pour plate method to observe the total bacteria counts in used medical face masks. The bacterial community analysis was detected using bio-Mass spectrometry technology and 16SrRNA gene sequencing technology. The wearing time of the mask from 0.5 hours to 5 hours were studied. These results shown that the total number of bacteria on the inside surface of the mask were higher than the outside. The total number of bacteria on the inner surface of masks worn for 0.5 h, 1 h 2 h, 4 h and 5 h was 69 CFU/m2,91.3 CFU/m2, 159.6 CFU/m2, 219 CFU/m2, and 879 CFU/m2, respectively. The total number of bacteria on the outside surface of masks worn for 0.5 h, 1 h 2 h, 4 h and 5 h was 60 CFU/m2, 82.7 CFU/m2, 119.8 CFU/m2, 200 CFU/m2, and 498 CFU/m2, respectively. The bacterial abundance obtained from bio-Mass spectrometry were consistent with the results of 16SrRNA sequencing. Both the methods discovered the maximum number of Neisseria followed by Corynebacterium species in mask worn 5 hours. The top 100 bacteria isolated from inside and outside surface of mask belong to 11 phyla. We analyzed bacterial penetration efficiency of the bacteria that were detected both on the inside and outside surface of the masks. In the top 10 bacteria, no bacteria were detected both inside and outside the mask worn for four hours, while 6 bacteria species were detected on the inside and outside of the mask after wearing for five hours. Bacterial penetration rates ranged from 0.74% to 99.66% for masks worn continuously for five hours, and the penetration rate of four strains exceeded 10% in the top 10 colonies. We recommend timely replacement of masks worn for more than four hours.

Pathogenic bacteria exploit transferrin receptor transcytosis to penetrate the blood–brain barrier

The human blood-brain barrier (BBB) comprises a single layer of brain microvascular endothelial cells (HBMECs) protecting the brain from bloodborne pathogens. Meningitis is among the most serious diseases, but the mechanisms by which major meningitis-causing bacterial pathogens cross the BBB to reach the brain remain poorly understood. We found that Streptococcus pneumoniae, group B Streptococcus, and neonatal meningitis Escherichia coli commonly exploit a unique vesicle fusion mechanism to hitchhike on transferrin receptor (TfR) transcytosis to cross the BBB and illustrated the details of this process in human BBB model in vitro and mouse model. Toll-like receptor signals emanating from bacteria-containing vesicles (BCVs) trigger K33-linked polyubiquitination at Lys168 and Lys181 of the innate immune regulator TRAF3 and then activate the formation of a protein complex containing the guanine nucleotide exchange factor RCC2, the small GTPase RalA and exocyst subcomplex I (SC I) on BCVs. The distinct function of SEC6 in SC I, interacting directly with RalA on BCVs and the SNARE protein SNAP23 on TfR vesicles, tethers these two vesicles and initiates the fusion. Our results reveal that innate immunity triggers a unique modification of TRAF3 and the formation of the HBMEC-specific protein complex on BCVs to authenticate the precise recognition and selection of TfR vesicles to fuse with and facilitate bacterial penetration of the BBB.

In vitro Comparative Evaluation of Polylactic Acid/Polyglycolic Acid Copolymer and Expanded Polytetrafluoroethylene Based Guided Tissue Regeneration Membrane for Bacterial Colonisation

The study was designed to compare the extent of bacterial colonisation on the surface of Polylactic acid/Polyglycolic acid copolymer and expanded Polytetrafluoroethylene based Guided tissue regeneration (GTR) membrane in an in vitro model by spot analysis and scanning electron microscopy. Earlier in vitro models have aimed to study the barrier function against the bacterial penetration across just one surface of the GTR membranes. No such study is present in the literature which aimed to quantify bacterial adhesion over both the surfaces of the membrane. Sterile Resorbable PLA/PGA copolymer and non-resorbable microporous ePTFE based GTR membrane were used in the study. Both groups were subdivided into two subgroup (n=10) based on incubation period of 24 and 48 hours. Incubated in Todd Hewitt broth with Streptococcus mutans, the samples were vortexed and analysed for bacterial count using spot test and scanning electron microscopy. Between PLA/PGA and ePTFE GTR membrane at 24 hours incubation time period, there was a significant difference in terms of Colony forming units (p = <0.001), with the median Colony forming units being highest in the PLA/PGA GTR membrane. At 48 hours incubation time period, there was a significant difference in terms of Colony forming units (p = <0.001), with the median Colony forming units being highest in the PLA/PGA GTR membrane. Our findings depict that ePTFE based GTR membrane showed significantly lesser bacterial colonisation on its surfaces as compared to PLA/PGA at both the incubation periods i.e., 24 hours and 48 hours as shown by both spot test and SEM.

Dual RNA-seq identifies genes and pathways modulated during Clostridioides difficile colonization.

The gastrointestinal pathogen Clostridioides difficile is the most common cause of hospital-acquired diarrhea. Bacterial interactions with the gut mucosa are crucial for the establishment of C. difficile infection; however, key infection events like bacterial attachment and gut penetration are still poorly defined. To better understand the initial events that occur when this anaerobe interacts with the human gut epithelium, we employed a dual RNA-sequencing approach to study the bacterial and host transcriptomic profiles during C. difficile infection in a dual environment in vitro human gut model. Temporal changes in gene expression during infection were studied in bacterial and epithelial cells over 3-24 hours. While there were several common differentially expressed bacterial genes across different timepoints after infection, mammalian transcriptional profiles were quite distinct, with little overlap. Interestingly, an induction of colonic receptors for C. difficile toxins was observed, along with the downregulation of genes encoding immune response markers. Several cell wall-associated proteins were downregulated in C. difficile when associated with host cells, including slpA, which encodes the main S-layer protein. Gene function and pathway enrichment analyses revealed a potential modulation of the purine/pyrimidine synthesis pathways both in the mammalian and bacterial cells. We observed that proline-proline endopeptidase, a secreted metalloprotease responsible for cell surface protein cleavage, is downregulated during infection, and a mutant lacking this enzyme demonstrated enhanced adhesion to epithelial cells during infection. This study provides new insight into the host and bacterial pathways based on gene expression modulation during the initial contact of C. difficile with gut cells. IMPORTANCE The initial interactions between the colonic epithelium and the bacterium are likely critical in the establishment of Clostridioides difficile infection, one of the major causes of hospital-acquired diarrhea worldwide. Molecular interactions between C. difficile and human gut cells have not been well defined mainly due to the technical challenges of studying cellular host-pathogen interactions with this anaerobe. Here we have examined transcriptional changes occurring in the pathogen and host cells during the initial 24 hours of infection. Our data indicate several changes in metabolic pathways and virulence-associated factors during the initial bacterium-host cell contact and early stages of infection. We describe canonical pathways enriched based on the expression profiles of a dual RNA sequencing in the host and bacterium, and functions of bacterial factors that are modulated during infection. This study thus provides fresh insight into the early C. difficile infection process.

Antibiofilm and antimicrobial activity of temporary filling materials on root canals: an in situ acid challenge.

The present study evaluated the antibiofilm and antimicrobial effects of temporary restorative materials on root canals after an intra-oral challenge. Seventy roots were endodontically treated and divided into 5 groups: high-viscosity glass ionomer (HV-GIC), light-activated glass ionomer (RM-GIC), zinc-oxide cement without eugenol (ZO), zinc-oxide cement with eugenol (ZOE), and unsealed roots (negative control). For 28 days, 14 participants used intra-oral devices with five roots, and drops of sucrose were applied onto them. The amount of biofilm and the bacterial counts were analyzed by Kruskal-Wallis and Dunn, and by two-way ANOVA and Tukey (α = 0.05). HV-GIC and RM-GIC better inhibit biofilm, followed by ZO and ZOE. Unsealed roots had the largest biofilm accumulation (p = 0.002) and higher bacterial penetration than restored roots (p = 0.023). A low amount of Streptococcus was found in RM-GIC and ZOE-restored roots without difference from HV-GIC (p = 0.021). The low amount of Enterococcus (p = 0.003) was found in the ZOE-restored roots, without difference from GICs.

An Ex-Vivo Model for Investigating Bacterial Extrusion from Infected Root Canals during Masticatory Function

IntroductionThe extrusion of bacteria from infected root canals may lead to increase in symptoms, expansion of periapical lesions, and contribution to systemic diseases. The aim of this study is to investigate a potential proof-of-concept model to study the extent to which bacteria can escape from infected root canals under dynamic loading (simulated chewing). MethodsThe study was completed in 2 experiments performed at 2 institutions. Biofilms of Streptococcus intermedius in the first experiment and S. intermedius and Actinomyces naeslundii were allowed to grow in root canals of single-rooted extracted teeth for 3 weeks. The roots of the teeth were suspended in a small chamber containing dental transport medium and were mounted on a lower sample holder of a chewing simulator. In the experimental group, simulated chewing cycles equivalent to 1 year of function were conducted, and then bacterial migration was quantified and compared with stationary teeth. ResultsAll experimental samples of the loading group revealed bacterial penetration in both experiments. Several of the unloaded samples revealed no bacterial penetration. In the first experiment, a significantly higher number of bacteria were able to escape into the periapex of the loaded group compared with the unloaded group (P = .017). In the second experiment, there was no significant difference between the 2 bacterial species used in the amount of extruded bacteria; however, there was a highly significant effect for occlusal loading (P = .0001). ConclusionsThe potential for occlusal forces to enhance bacterial extrusion from infected root canals should be further explored.

Microleakage of Restorative Materials Used for Temporization of Endodontic Access Cavities.

A tight temporary seal applied to an access cavity is thought to improve endodontic outcomes. This study aims to assess the bacterial and glucose microleakage of different types and combinations of temporary restorations. Human-extracted incisors were instrumented, dressed with a calcium hydroxide paste, and sealed with Cavit W (CW), CW/Ketac Molar (CW/KM), CW/Smart Dentin Replacement (CW/SDR), Intermediate restorative material/KM (IRM/KM), or Clip F (CF). Standardized 3D-printed hollow test specimens were manufactured and temporized in the same manner. The specimens were examined for bacterial and glucose leakage for 28 days. Data were analyzed using a Kaplan-Meier survival analysis. CW/SDR and CF showed the least bacterial and glucose leakage over time. CW, CW/KM, and IRM/KM had similarly high levels of glucose leakage, but CW/KM and IRM/KM provided a tighter seal against bacterial penetration than CW. CW/SDR and CF should be considered for the sealing of access cavities of teeth previously restored with methacrylate-based materials.

Development of light-emitting Episcia lilacina leaf by applying Vibrio campbellii RMT1 and extending the glowing by CaCl2 and yeast extract.

Glowing Episcia lilacina was generated through foliar application of the bioluminescent bacterium Vibrio campbellii RMT1. Firstly, different nutrient formulas were tested, incorporating yeast extract and various inorganic salts, such as CaCl2, MgCl2, MgSO4, KH2PO4, K2HPO4, and NaCl, in order to enhance bacterial growth and light emission. The combination of 0.15% of yeast extract and 0.3% of CaCl2 in a nutrient broth (NB) + 1% NaCl medium extended light emission to 24h and resulted in higher light intensity compared to other combinations of yeast extract and inorganic salts. The peak intensity reached approximately 1.26 × 108 relative light units (RLU) at 7h. The optimal presence of inorganic salt ions likely contributed to enhanced light emission, while the yeast extract acted as a nutrient source. Secondly, the effect of proline on salt-induced stress symptoms was investigated by applying 20mM proline to the glowing plant. Additionally, a 0.5% agar nutrient was spread on the leaves prior to bacteria application to support bacterial growth and penetration. Exogenous proline application led to a significant accumulation of proline in plant cells, resulting in decreased malondialdehyde (MDA) levels. However, the proline accumulation also reduced the light intensity of the bioluminescent bacteria. This study demonstrates the potential for generating light on a living plant using bioluminescent bacteria. Further understanding of the interaction between plants and light-emitting bacteria could contribute to the development of sustainably light-emitting plants.

Development of antisense peptide-peptide nucleic acids against fluoroquinolone-resistant Escherichia coli.

Fluoroquinolones (FQs) are potent and broad-spectrum antibiotics commonly used to treat MDR bacterial infections, but bacterial resistance to FQs has emerged and spread rapidly around the world. The mechanisms for FQ resistance have been revealed, including one or more mutations in FQ target genes such as DNA gyrase (gyrA) and topoisomerase IV (parC). Because therapeutic treatments for FQ-resistant bacterial infections are limited, it is necessary to develop novel antibiotic alternatives to minimize or inhibit FQ-resistant bacteria. To examine the bactericidal effect of antisense peptide-peptide nucleic acids (P-PNAs) that can block the expression of DNA gyrase or topoisomerase IV in FQ-resistant Escherichia coli (FRE). A set of antisense P-PNA conjugates with a bacterial penetration peptide were designed to inhibit the expression of gyrA and parC and were evaluated for their antibacterial activities. Antisense P-PNAs, ASP-gyrA1 and ASP-parC1, targeting the translational initiation sites of their respective target genes significantly inhibited the growth of the FRE isolates. In addition, ASP-gyrA3 and ASP-parC2, which bind to the FRE-specific coding sequence within the gyrA and parC structural genes, respectively, showed selective bactericidal effects against FRE isolates. Our results demonstrate the potential of targeted antisense P-PNAs as antibiotic alternatives against FQ-resistance bacteria.

PH/Hyal-Responsive Surface-Charge Switchable Electrostatic Complexation for Efficient Elimination of MRSA Infection.

Methicillin-resistant Staphylococcus aureus (MRSA) has become a great threat to human health worldwide, making new effective antibacterial strategies urgently desired. In this study, a cationic pH-responsive delivery system (pHSM) was developed based on poly(β-amino esters)-methoxy poly(ethylene glycol), by which linezolid (LZD) could be encapsulated to form pHSM/LZD. The biocompatibility and stability of pHSM/LZD were further enhanced by adding low-molecular-weight hyaluronic acid (LWT HA) on the surface through electrostatic interaction to form pHSM/LZD@HA, of which the positive surface charges were neutralized by LWT HA under physiological conditions. LWT HA can be degraded by hyaluronidase (Hyal) after arriving at the infection site. In vitro, pHSM/LZD@HA could rapidly change to being positively charged on the surface within 0.5 h under acidic conditions, especially when Hyal was present, thus promoting bacterial binding and biofilm penetration of pHSM/LZD@HA. In addition, the pH/Hyal-dependent accelerated drug release behavior was also observed and it is beneficial for the comprehensive treatment of MRSA infection in vitro and in vivo. Our study provides a novel strategy to develop a pH/Hyal-responsive drug delivery system for the treatment of MRSA infection.

Porphyromonas gingivalis Gingipains Destroy the Vascular Barrier and Reduce CD99 and CD99L2 Expression To Regulate Transendothelial Migration.

Porphyromonas gingivalis is an important periodontal pathogen that can cause vascular injury and invade local tissues through the blood circulation, and its ability to evade leukocyte killing is critical to its distal colonization and survival. Transendothelial migration (TEM) is a series of that enable leukocytes to squeeze through endothelial barriers and migrate into local tissues to perform immune functions. Several studies have shown that P. gingivalis-mediated endothelial damage initiates a series of proinflammatory signals that promote leukocyte adhesion. However, whether P. gingivalis is involved in TEM and thus influences immune cell recruitment remains unknown. In our study, we found that P. gingivalis gingipains could increase vascular permeability and promote Escherichia coli penetration by downregulating platelet/endothelial cell adhesion molecule 1 (PECAM-1) expression in vitro. Furthermore, we demonstrated that although P. gingivalis infection promoted monocyte adhesion, the TEM capacity of monocytes was substantially impaired, which might be due to the reduced CD99 and CD99L2 expression on gingipain-stimulated endothelial cells and leukocytes. Mechanistically, gingipains mediate CD99 and CD99L2 downregulation, possibly through the inhibition of the phosphoinositide 3-kinase (PI3K)/Akt pathway. In addition, our in vivo model confirmed the role of P. gingivalis in promoting vascular permeability and bacterial colonization in the liver, kidney, spleen, and lung and in downregulating PECAM-1, CD99, and CD99L2 expression in endothelial cells and leukocytes. IMPORTANCE P. gingivalis is associated with a variety of systemic diseases and colonizes in distal locations in the body. Here, we found that P. gingivalis gingipains degrade PECAM-1 to promote bacterial penetration while simultaneously reducing leukocyte TEM capacity. A similar phenomenon was also observed in a mouse model. These findings established P. gingivalis gingipains as the key virulence factor in modulating the permeability of the vascular barrier and TEM processes, which may provide a new rationale for the distal colonization of P. gingivalis and its associated systemic diseases.

Development of breathable and liquid/wet bacterial penetration barrier composite laminated fabrics for surgical gown applications

Disposable surgical gowns are used to protect health care workers from microorganisms and body fluids to meet the needs of surgical applications. In addition to protection, it has a significant impact on the surgical team members’ comfort and thus, the success of the operations. In this study, a dual layer surgical gown-an outer layer of film and an inner layer of nonwoven fabric-was developed. Produced composite laminated fabrics are SMS (spunbond/meltblown/spunbond) nonwoven fabric laminated microporous polyethylene film and SMS (spunbond/meltblown/spunbond) nonwoven fabric laminated thermoplastic polyurethane films. These fabrics were compared with nonporous polyethylene film-coated SS (spunbond/spunbond) nonwoven fabric available in the local market. The study aims to investigate the effect of some structural factors of composite nonwoven laminated fabrics, including lamination process, manufacturing process, and film type; microporous or monolithic, nonwoven fabric manufacturing process, and helium atmospheric-pressure glow discharge plasma treatment, on their functional performance properties, including air permeability, water vapour permeability, hydrostatic pressure resistance, and wet bacterial penetration resistance. The results showed that the produced composite fabrics offer outstanding breathability and, thus, the surgical team members' comfort. Produced composite fabrics had no wet bacterial penetration of Staphylococcus aureus and thus contributed to the prevention of the transfer of bacterial pathogens to prevent surgical site infection or post-operative infection. Produced composite fabrics offer superior liquid barriers according to the barrier protection classification of the Association for the Advancement of Medical Instrumentation (ANSI/AAMI PB 70:12) and thus contribute to the prevention of the transfer of body fluids during surgical operations.

Effects of Organic Ligands on the Antibacterial Activity of Reduced Iron-Containing Clay Minerals: Higher Extracellular Hydroxyl Radical Production Yet Lower Bactericidal Activity.

Reduced iron-containing clay (RIC) minerals have been documented to exhibit antibacterial activity through a synergistic action of extracellular membrane attack and intracellular oxidation of cellular components. However, the relative importance between extracellular and intracellular processes has remained elusive. Here, metal-chelating organic ligands (lactate, oxalate, citrate, and ethylene diaminetetraacetic acid (EDTA)) were amended to the bactericidal assays such that the importance of the two processes could be evaluated. Reduced nontronite (rNAu-2) was used as a model clay mineral to produce extracellular hydroxyl radical (•OH) upon oxygenation. The presence of Fe-chelating ligands increased •OH yield by 3-5 times. Consequently, bacterial cell membrane attack was enhanced, yet the antibacterial activity of RIC diminished. Additional experiments revealed that the ligands inhibited soluble metal ions from adsorption onto the bacterial cell membrane and/or penetration into the cytoplasm. Consequently, intracellular Fe concentration for the ligand-treated group was nearly 2 orders of magnitude lower than that for no-ligand control, which greatly decreased intracellular accumulation of reactive oxygen species (ROS) and increased cell survival. These results highlight that destruction of intracellular contents (proteins and DNA) is more important than oxidative degradation of membrane lipids and cell envelope proteins in causing bacterial cell death by RIC.

Edaravone-Delivering PVA-Chitosan Nanofibrous Wound Dressings Promote Diabetic Wound Healing Through Modulation of Oxidative Stress and Inflammation

In the current study, edaravone was loaded into electrospun PVA/chitosan wound dressings to develop a potential wound care material for diabetic wounds. Different in vitro studies were performed to characterize the dressings. In vivo study was carried out in a rat model of diabetic wound healing. Study showed that the nanofibrous membranes prevented bacterial penetration and showed no toxicity toward skin fibroblast cells. In vivo study showed that edaravone-loaded wound dressings had the highest rate of wound closure and formation of new epithelium compared with other groups. Gene expression study showed that TNF-a and GP-x genes tissue expression was significantly downregulated in the rats treated with edaravone-delivering wound dressings, implying that these dressings have modulated oxidative stress and inflammation in diabetic wounds.

Bacterial leaf spot of Ilex paraguariensis caused by Pseudomonas syringae

AbstractYerba mate (Ilex paraguariensis) leaves and twigs are dried and used to make mate, a popular herbal tea in Southern Brazil, Argentina, Paraguay and Uruguay. In February 2021, yerba mate leaves with water‐soaked spots, surrounded by chlorotic halos were observed in four nurseries at the Ilópolis and Arvorezinha counties, Rio Grande do Sul (RS) State, Brazil. Bacterial colonies were recovered from the symptomatic leaves. Based on morphological characteristics, LOPAT tests and the DNA sequence of four house‐keeping genes, the causal agent was identified as Pseudomonas syringae. To fulfil Koch's postulates, pathogenicity was confirmed by inoculating leaves of yerba mate seedlings with a bacterial suspension obtained from one of the isolates, under laboratory conditions. Inoculated leaves developed small dark spots after 4 days post‐inoculation only in plants that were wounded prior to the inoculation, indicating that mechanical disruption of the plant tissue favoured bacterial penetration. To the best of our knowledge, this is the first worldwide report of P. syringae causing bacterial leaf spots on yerba mate.

Bacterial Colonization and Proliferation in Primary Molars following the Use of the Hall Technique: A Confocal Laser Scanning Microscopy Study.

Restorative dentistry aims to create a favorable environment to arrest caries with minimal operative intervention. The Hall technique (HT) involves the seating and cementation of stainless steel crowns (SSC) on primary molars without any tooth preparation, caries removal, or local anesthesia. In this manner, it entombs bacteria and arrests caries' progress. We compared bacterial distribution and quantity among primary molars affected with caries and restored with SSC using the HT (n = 10), the conventional technique (CT; n = 10), or not restored at all (control; n = 10). The teeth were contaminated with Enterococcus faecalis to mimic the clinical situation in the oral cavity and then incubated for 21 days. They were then cut mesiodistally and evaluated with confocal laser scanning microscopy. Total bacterial load (live + dead) in the mesial and distal areas of the crown showed no significant difference between the groups (p = 0.711), but there were significantly more dead than live bacteria in the CT and control groups versus the HT group (p = 0.0274 and p = 0.0483, respectively). Inside the pulp chamber and the crown area, the total bacterial load was significantly higher in the HT compared to the CT group (p < 0.001). Significantly more dead than live bacteria were observed in all tooth areas treated with the HT (p = 0.0169). Bacterial penetration depth was significantly correlated with bacterial load (p = 0.0167). In conclusion, although more bacteria were present in teeth that had undergone the HT versus those treated with the CT, they were mainly unviable. Additionally, the CT and the HT showed a similar performance in terms of marginal leakage, indicating that complete caries removal is not essential to achieve good sealing.