Biofilm Integrity Research Articles

E. coli cells advance into phase-separated (biofilm-simulating) extracellular polymeric substance containing DNA, HU, and lipopolysaccharide.

Understanding the constitution and behavior of biofilms is crucial to understanding how to deal with persistent biofilms. This study, together with other recent studies from our group, elucidates a novel aspect of the extracellular polymeric substance (EPS) of Escherichia coli biofilms, by creating a simulacrum of the EPS and then demonstrating that its formation involves liquid-liquid phase separation (LLPS) of HU, DNA, and lipopolysaccharide (LPS) components, with LPS determining the liquidity of this EPS simulacrum. The findings provide insight into the nature of biofilms and into how the interplay of HU, DNA, and LPS could modulate the structural integrity and functional dynamics of biofilms.

Effect of wastewater containing mixture of nanoparticles on organic matter removal, power generation, and biofilm integrity in sediment microbial fuel cell

ABSTRACT Industrial wastewaters from the cosmetic industry contain high organic strength and mixture of nanoparticles (NPs). Sediment microbial fuel cell (MFC) is an emerging nature-based technology that can treat complex wastewaters. The aim of this study is to understand the effect of a binary mixture of zinc oxide (ZnO) and copper oxide (CuO) NPs (concentration: 1 + 1 and 10 + 10 mg/L) on the organic matter removal, power generation, and biofilm health of sediment MFCs after a long-term operation of 120 days. The high chemical oxygen demand (COD) removal (>95%) observed for all reactors signified the minimal impact of 10 mg/L NP mixture on treatment. The dissolved organic carbon (DOC) removal from the sediment was reduced by 8% due to NPs. NPs also led to 42.2% higher anode extracellular polymeric substances (EPS) and 46.65% lesser cathode EPS generation. The maximum power density of 0.29 mW/m2 was obtained for the 10 mg/L NP reactor, with the average being 23% higher than the no-NP control reactor. This was the first study to explore the effect of the mixture of NPs on the performance of an MFC. The results indicated that sediment MFCs can sustain high mixture concentrations of NPs. Furthermore, variation of parameters can aid in establishing the feasibility of this technology for treating wastewater with NPs.

A pH-modulated polymeric drug-release system simultaneously combats biofilms and planktonic bacteria to eliminate bacterial biofilm

Bacterial infection, leading to biofilm formation, raises grave concerns. With the aim of eliminating both integrity of biofilms and individual bacterial cells, it is desirable to develop bacterial biofilm-eliminating material to be combined with dispersal and anti-microbial agents. Herein, a biofilm-eliminating enzyme (α amylase) was incorporated in novel pH-sensitive polymer hydrogel, Poly (methacrylic acid-co-2-(dimethylamino)ethyl methacrylate), which encapsulates drug-loaded porous Zeolitic Imidazolate Framework-8 (ZIF-8). At body temperature (37 °C) the maximum swelling of the hydrogel was observed at pH 8.5, which assisted in rupturing biofilm by releasing α amylase. Amylase, encapsulated in polymer matrix reached 90% release after 6 days at 8.5, while CIP was released 96% on day 23 at pH 8.5. The antibiofilm activity of drug-releasing system was assessed against E. coli and S. aureus. The overall bacterial viability of biofilms showed significant time-dependent declines, when exposed to drug-loaded hydrogel. Importantly, cytotoxicity experiments showed excellent biocompatibility of hydrogel. In the in vivo deep wound infection model, drug-loaded hydrogel treatment showed complete wound closure with proper tissue regeneration. No inflammatory, or systemic toxicity of hydrogel was observed. The drug release from hydrogel is initiated only upon receiving physiological cue arising due to biofilm formation. Therefore, this hydrogel serves as reservoir for antibacterial drug, which release it only in presence of biofilm and thus work for sustained release for long period, compared to conventional drug loaded films or coatings. A powerful modality for management of biofilm-associated infections can be provided by the proposed synergistic strategy for complete elimination of biofilms.

Study of chlorine disinfection on the formation and resuscitation of Viable but nonculturable (VBNC) bacteria in drinking water distribution systems

The presence of Viable but nonculturable (VBNC) bacteria in drinking water distribution systems(DWDS)which can recover and regain pathogenicity under certain conditions, leads to a serious underestimation of the number of microorganisms in the water and consequently microbiological risks. This study investigated E. coli's biofilm integrity and metabolic activity post‑chlorine disinfection and during resuscitation in a Circulating-flow Dynamic biofilm Contactor (CDC) reactor. Chlorine rapidly induced the VBNC state, with concentrations ≥0.3 mg/L causing all respiratory-active bacteria to transition. Higher chlorine levels damaged cell biofilms, reducing intact cell counts by 95 % and depleting ATP. VBNC bacteria in biofilms quickly regained culturability, utilizing available nutrients and ATP. Within 1–4 days, culturable bacteria in the biofilm increased from 2.29 to 4.95 log CFU/mL, significantly faster than in water. This growth corresponded with rising ATP levels, though slower than in aqueous environments. Signaling molecules (C6, C8, C12, C14) regulated gene expression during resuscitation, aiding bacterial recovery and proliferation. Chlorine disinfection disrupted intercellular communication, altering VBNC bacteria's physiological and metabolic characteristics. These findings highlight the complex dynamics of VBNC bacteria in water systems, their resuscitation potential, and the importance of biofilms in bacterial recovery, emphasizing the need for comprehensive water treatment strategies.

Strain of Xanthomonas oryzae pv. oryzae Loses Virulence through Dysregulation of Cardiolipin Synthase.

Small non-coding RNAs (sRNAs) are pivotal post-transcriptional regulatory factors influencing biological activity. Studies on the rice bacterial blight pathogen Xanthomonas oryzae pathovar oryzae strain PXO99A, previously identified a virulence-associated sRNA, trans3287. A mutant strain lacking this sRNA, named SK01, resulted in markedly diminished virulence towards rice. This study aims to further elucidate the underlying bacterial virulent function of trans3287. The expression of trans3287 was quantified in virulence-inducing and standard nutritional conditions to clarify its production mechanism. The detection of virulence-associated genes revealed that trans3287 regulated the synthesis processes of extracellular polysaccharides, lipopolysaccharides, and the type III secretion system. Moreover, bioinformatics prediction and quantitative PCR indicated a potential direct target of trans3287, PXO_03470, encoding cardiolipin synthase. A dual-plasmid system fusing with GFP tag and protein immunoblotting confirmed that sRNA trans3287 negatively regulated PXO_03470. Bacterial biofilms demonstrated trans3287 regulated the disruption of biofilm integrity through cardiolipin synthase. This study provides preliminary insights into the mechanistic underpinnings of the role of sRNA trans3287 in mediating bacterial virulence through cardiolipin synthase.

Spatial propagation of temperate phages within and among biofilms.

Bacteria form groups comprised of cells and a secreted polymeric matrix that controls their spatial organization. These groups - termed biofilms - can act as refuges from environmental disturbances and from biotic threats, including phages. Despite the ubiquity of temperate phages and bacterial biofilms, live propagation of temperate phages within biofilms has never been characterized on cellular spatial scales. Here, we leverage several approaches to track temperate phages and distinguish between lytic and lysogenic host infections. We determine that lysogeny within E. coli biofilms initially occurs within a predictable region of cell group packing architecture on the biofilm periphery. Because lysogens are generally found on the periphery of large cell groups, where lytic viral infections also reduce local biofilm cell packing density, lysogens are predisposed to disperse into the passing liquid and are over-represented in biofilms formed from the dispersal pool of the original biofilm-phage system. Comparing our results with those for virulent phages reveals that temperate phages have previously unknown advantages in propagating over long spatial and time scales within and among bacterial biofilms.

Membrane protein Bcsdr2 mediates biofilm integrity, hyphal growth and virulence of Botrytis cinerea

Grey mould caused by Botrytis cinerea is a devastating disease responsible for large losses to agricultural production, and B. cinerea is a necrotrophic model fungal plant pathogen. Membrane proteins are important targets of fungicides and hotspots in the research and development of fungicide products. Wuyiencin affects the permeability and pathogenicity of B. cinerea, parallel reaction monitoring revealed the association of membrane protein Bcsdr2, and the bacteriostatic mechanism of wuyiencin was elucidated. In the present work, we generated and characterised ΔBcsdr2 deletion and complemented mutant B. cinerea strains. The ΔBcsdr2 deletion mutants exhibited biofilm loss and dissolution, and their functional activity was illustrated by reduced necrotic colonisation on strawberry and grape fruits. Targeted deletion of Bcsdr2 also blocked several phenotypic defects in aspects of mycelial growth, conidiation and virulence. All phenotypic defects were restored by targeted gene complementation. The roles of Bcsdr2 in biofilms and pathogenicity were also supported by quantitative real-time RT-PCR results showing that phosphatidylserine decarboxylase synthesis gene Bcpsd and chitin synthase gene BcCHSVII were downregulated in the early stages of infection for the ΔBcsdr2 strain. The results suggest that Bcsdr2 plays important roles in regulating various cellular processes in B. cinerea.Key points• The mechanism of wuyiencin inhibits B. cinerea is closely associated with membrane proteins.• Wuyiencin can downregulate the expression of the membrane protein Bcsdr2 in B. cinerea.• Bcsdr2 is involved in regulating B. cinerea virulence, growth and development.

Design of Ultrasound-Driven Charge Interference Therapy for Wound Infection.

Wound infections, especially those caused by pathogenic bacteria, present a considerable public health concern due to associated complications and poor therapeutic outcomes. Herein, we developed antibacterial nanoparticles, namely, PGTP, by coordinating guanidine derivatives with a porphyrin-based sonosensitizer. The synthesized PGTP nanoparticles, characterized by their strong positive charge, effectively disrupted the bacterial biosynthesis process through charge interference, demonstrating efficacy against both Gram-negative and Gram-positive bacteria. Additionally, PGTP nanoparticles generated reactive oxygen species under ultrasound stimulation, resulting in the disruption of biofilm integrity and efficient elimination of pathogens. RNA-seq analysis unveiled the detailed mechanism of wound healing, revealing that PGTP nanoparticles, when coupled with ultrasound, impair bacterial metabolism by interfering with the synthesis and transcription of amino acids. This study presents a novel approach to combatting wound infections through ultrasound-driven charge-interfering therapy, facilitated by advanced antibacterial nanomaterials.

Methods for Fixing Biofilms of Staphylococcus aureus and Salmonella enterica for Microscopic Examination.

Different methods for fixing biofilms of Staphylococcus aureus and Salmonella enterica for light and electron microscopy were compared. Paraformaldehyde fixation did not preserve biofilm integrity during dehydration; Ito-Karnovsky fixation revealed cell morphology, but did not preserve the matrix. Ruthenium red combined with aldehydes allowed the matrix to be preserved and visualized. An analysis of the ultrastructure of S. aureus and S. enterica cells in biofilms and suspensions at various fixations is presented. The ultrastructure of the biofilm matrix has been described.

Mucus and Biofilm Penetrating Nanoplatform as an Ultrasound-Induced Free Radical Initiator for Targeted Treatment of Helicobacter pylori Infection.

Helicobacter pylori (H. pylori) infection is closely associated with the development of various gastric diseases. The effectiveness of current clinical antibiotic therapy is hampered by the rise of drug-resistant strains and the formation of H. pylori biofilm. This paper reports a sonodynamic nanocomposite PtCu3-PDA@AIPH@Fucoidan (PPAF), which consists of dopamine-modified inorganic sonosensitizers PtCu3, alkyl radicals (R•) generator AIPH and fucoidan, can penetrate the mucus layer, target H. pylori, disrupt biofilms, and exhibit excellent bactericidal ability. In vitro experiments demonstrate that PPAF exhibits excellent acoustic kinetic properties, generating a significant amount of reactive oxygen species and oxygen-independent R• for sterilization under ultrasound stimulation. Simultaneously, the produced N2 can enhance the cavitation effect, aiding PPAF nanoparticles in penetrating the gastric mucus layer and disrupting biofilm integrity. This disruption allows more PPAF nanoparticles to bind to biofilm bacteria, facilitating the eradication of H. pylori. In vivo experiments demonstrate that ultrasound-stimulated PPAF exhibited significant antibacterial efficacy against H. pylori. Moreover, it effectively modulated the expression levels of inflammatory factors and maintained gastrointestinal microbiota stability when compared to the antibiotic treatment group. In summary, PPAF nanoparticles present a potential alternative to antibiotics, offering an effective and healthy option for treating H. pylori infection.

Purification, characterization, and identification of a novel bacteriocin produced by Lacticaseibacillus casei KLS1, and its antimicrobial mechanism against Staphylococcus aureus

Staphylococcus aureus (S. aureus) and its biofilm were recognized as one of the major threats to food safety and human health. Bacteriocins produced by Lacticaseibacillus species have been demonstrated to inhibit the growth of foodborne pathogenic bacteria and their biofilm formation. Herein, a novel bacteriocin KLS1 from Lacticaseibacillus casei KLS1 was extracted by ethyl acetate and purified using an AKTA Pure 25 protein chromatography purification system. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses revealed that the molecular mass of bacteriocin KLSI was 3786.91 Da and the amino acid sequence was DTSASIASNKSETNDLLKQIEAANTEVINLNKQID. Bacteriocin KLS1 was stable at pH 2.0–6.0, sensitive to proteases, and showed excellent thermal stability. Furthermore, bacteriocin KLS1 exhibited broad-spectrum antimicrobial activity against both Gram-positive and Gram-negative bacteria, even fungi. The crystal violet staining assay indicated that bacteriocin KLS1 significantly reduced the relative percentage of S. aureus biofilm (P < 0.05). Meanwhile, bacteriocin KLS1 was found to significantly decrease the metabolic activity of S. aureus by Cell Counting Kit-8 assay (P < 0.05). In addition, scanning electron microscopy and confocal laser scanning microscope showed that bacteriocin KLS1 disrupted the structure of S. aureus biofilm and cell membrane integrity, leading to cell deformation and leakage of contents. These results suggested that bacteriocin KLS1 might have the potential to serve as a biological preservative in the food industry.

Exploring Citronella's inhibitory mechanism against Listeria monocytogenes and its utilization in preserving cheese

Listeria monocytogenes presents significant risk to human health due to its high resistance and capacity to form toxin-producing biofilms that contaminate food. The objective of this study was to assess the inhibitory effect of citronella aldehyde (CIT) on L. monocytogenes and investigate the underlying mechanism of inhibition. The results indicated that the minimum inhibitory concentration (MIC) and Minimum sterilisation concentration (MBC) of CIT against L. monocytogenes was 2 μL/mL. At this concentration, CIT was able to effectively suppress biofilm formation and reduce metabolic activity. Crystalline violet staining and MTT reaction demonstrated that CIT was able to inhibit biofilm formation and reduce bacterial cell activity. Furthermore, the motility assessment assay revealed that CIT inhibited bacterial swarming and swimming. Scanning electron microscopy (SEM) and laser confocal microscopy (LSCM) observations revealed that CIT had a significant detrimental effect on L. monocytogenes cell structure and biofilm integrity. LSCM also observed that nucleic acids of L. monocytogenes were damaged in the CIT-treated group, along with an increase in bacterial extracellular nucleic acid leakage. The proteomic results also confirmed the ability of CIT to affect the expression of proteins related to processes including metabolism, DNA replication and repair, transcription and biofilm formation in L. monocytogenes. Consistent with the proteomics results are ATPase activity and ATP content of L. monocytogenes were significantly reduced following treatment with various concentrations of CIT. Notably, CIT showed good inhibitory activity against L. monocytogenes on cheese via fumigation at 4 °C.This study establishes a foundation for the potential application of CIT in food safety control.

Effect of alkali‐treated Putranjiva roxburghii seed shell filler on physico‐chemical, thermal, mechanical, and barrier properties of polyvinyl alcohol‐based biofilms

AbstractBiopolymers and bio‐fillers derived from natural, organic, and abundant resources have garnered more responsiveness owing to their affordability and degradability in the production of packaging plastics. This study explores the novel use of different proportions (5%, 10%, 15%, and 20%) of 5% alkali‐treated Putranjiva roxburghii seed shell filler (PRSSF) as a bio‐filler in combination with polyvinyl alcohol (PVA) for the first time. FTIR analysis showed the creation of robust hydrogen bonds and enhanced compatibility between the matrix and the alkalized PRSSF. The XRD results revealed that alkalized PRSSF strengthens the structural integrity of biofilms. The water absorption of the PVA/at PRSSF biofilm samples decreased by 88.38% at a higher composition (20%) of alkali‐treated PRSSF due to their alkalized hydrophobic filler. Due to the effect of at‐PRSSF into PVA, the resulting films demonstrated a degradation temperature and char residue of 334.8°C and 13.57%, respectively, and relatively better UV‐barrier properties in a range of visible light. When compared with pure PVA films, the tensile strength and corresponding modulus of PVA/20% at‐PRSSF films increased by 32.94% and 16.2%, respectively. Therefore, the PVA/at‐PRSSF biofilms produced in this study are ideal materials for wrapping and folding applications.Highlights PVA/at‐PRSSF films outperform in multiple aspects as compared with pure PVA. Tensile strength of PVA/20%‐PRSSF films increased by 32.94%. Elongation at break decreased with at‐PRSSF loading. Water absorption in PVA/20% PRSSF biofilm decreased by 88.38%. Fractography showed voids and agglomerations at high filler levels.

The antioxidant, antimicrobial and antibiofilm potential of biosurfactants derived from Enterococcus faecalisBHT‐2

AbstractBiosurfactants are amphiphilic molecules containing one hydrophilic and one hydrophobic moiety. They are synthesized by microorganisms either on the cell surface or secreted extracellularly which serve as natural surfactant substitutes with a wide range of applications in different sectors. The present study aims to assess the antioxidant, antimicrobial, and antibiofilm properties of biosurfactants derived from Enterococcus faecalis BHT‐2 against different microorganisms (E. clocae, P. aeruginosa, M. luteus, S. aureus, C. albicans, and E. coli). Derived biosurfactants showed antimicrobial activity against test microorganisms in the range between 0.6 and 3 mg/mL. It displayed potent antibiofilm ability by inhibiting the bacterial attachment to surfaces, leading to the disruption of biofilm formation by altering the integrity of microbial biofilms when observed on coverslips by SEM and fluorescence microscopy. The treatment of microbial biofilms by biosurfactants greatly reduced the exopolysaccharide biomass, that is, carbohydrate and protein content. Thus, in vitro assessment of antimicrobial and antibiofilm properties of biosurfactants derived from E. faecalis BHT‐2 indicated that it act as a better alternative antimicrobial agents.

Restraining Staphylococcus aureus Virulence Factors and Quorum Sensing through Lactic Acid Bacteria Supernatant Extracts.

The escalating prevalence of antibiotic-resistant bacteria poses a grave threat to human health, necessitating the exploration of novel alternatives to conventional antibiotics. This study investigated the impact of extracts derived from the supernatant of four lactic acid bacteria strains on factors contributing to the pathogenicity of three Staphylococcus aureus strains. The study evaluated the influence of lactic acid bacteria supernatant extracts on the growth, biofilm biomass formation, biofilm metabolic activity, and biofilm integrity of the S. aureus strains. Additionally, the impact on virulence factors (hemolysin and coagulase) was examined. Gas chromatography coupled with mass spectrometry was used to identify the bioactive compounds in the extracts, while molecular docking analyses explored potential interactions. Predominantly, the extracts contain eight 2,5-diketopiperazines, which are cyclic forms of peptides. The extracts demonstrated inhibitory effects on biofilm formation, the ability to disrupt mature biofilms, and reduce the biofilm cell metabolic activity of the S. aureus strains. Furthermore, they exhibited the ability to inhibit α-hemolysin production and reduce coagulase activity. An in silico docking analysis reveals promising interactions between 2,5-diketopiperazines and key proteins (SarA and AgrA) in S. aureus, confirming their antivirulence and antibiofilm activities. These findings suggest that 2,5-diketopiperazines could serve as a promising lead compound in the fight against antibiotic-resistant S. aureus.

Suppression of planktonic and biofilm of Escherichia coli by the synergistic lantibiotics-polymyxins combinations.

Escherichia coli are generally resistant to the lantibiotic's action (nisin and warnerin), but we have shown increased sensitivity of E. coli to lantibiotics in the presence of subinhibitory concentrations of polymyxins. Synergistic lantibiotic-polymyxin combinations were found for polymyxins B and M. The killing of cells at the planktonic and biofilm levels was observed for two collection and four clinical multidrug-resistant E. coli strains after treatment with lantibiotic-polymyxin B combinations. Thus, 24-h treatment of E. coli mature biofilms with warnerin-polymyxin B or nisin-polymyxin B leads to five to tenfold decrease in the number of viable cells, depending on the strain. AFM revealed that the warnerin and polymyxin B combination caused the loss of the structural integrity of biofilm and the destruction of cells within the biofilm. It has been shown that pretreatment of cells with polymyxin B leads to an increase of Ca2+ and Mg2+ ions in the culture medium, as detected by atomic absorption spectroscopy. The subsequent exposure to warnerin caused cell death with the loss of K+ ions and cell destruction with DNA and protein release. Thus, polymyxins display synergy with lantibiotics against planktonic and biofilm cells of E. coli, and can be used to overcome the resistance of Gram-negative bacteria to lantibiotics.

Enhancing manganese removal through pre-deposition of powdered activated carbon-manganese oxide (PAC-MnOx) film on gravity-driven ceramic membrane: The impact of biofilm integrity and fluid dynamics

Manganese is widely distributed in the natural environment, and its presence in drinking water poses a risk to human health. In this study, an integrated pre-deposited powdered activated carbon (PAC) and manganese oxide (MnOx) bio-functional layer of inorganic origin was coupled with gravity-driven ceramic membrane (GDCM) reactor. The study aims to investigate the effects of the membrane pore size and functional layer integrity on the Mn2+ removal and flux. The results reveal that the average Mn2+ removal efficiency of the large pore size membranes (300 kDa and 0.45 μm) was lower (75 % – 78 %) compared to those with smaller pores (15 kDa and 50 kDa) (87 % – 88 %) during long-term operation. Such differences are attributed to a period of manganese leakage before the PAC-MnOx matured in large-pore membrane units and a longer residence time in small-pore membranes. The functional layer leaded to reversible fouling, which was more prominent in membranes with larger pores (contributing to 93 % of total resistance) than those with smaller ones (ranging from 51.7 % to 61.9 %). Although the damaged functional layer resulted in occasional high Mn2+ concentrations in the effluent, it did not significantly impact long-term flux development. Optical coherence tomography (OCT), in conjunction with COMSOL analysis, non-destructively evaluate biofilm formation process and indicate a homogeneous flow field and mass transfer in the “dual layers” (active film layer and ceramic membrane layer) of the GDCM. The presence of manganese-oxidizing bacteria (Hyphomicrobium, Reyranella, and Pseudomonas) in the GDCM reactor was verified using confocal laser scanning microscopy (CLSM) and high-throughput sequencing. Additional material characterization revealed that a portion of the PAC-MnOx exhibited characteristics of birnessite, with autocatalytic oxidation of Mn2+, and that Mn (III) and Mn (IV) were the primary species in MnOx, which exhibited excellent catalytic oxidation and adsorption capabilities for Mn (II), respectively. Overall, the developed PAC-MnOx GDCM system shows great promise in efficiently removing manganese and producing water of desired quality, thereby holding potential in applications across several sectors of advanced water treatment processes.

Further Preclinical Development of a Bio‐therapeutic Against Porphyromonas gingivalis for the control of Systemic and Neuro‐inflammation related Dementia and Alzheimer’s Disease

AbstractAddressing novel mechanisms and effective therapeutic treatments for cognitive decline, Dementia and Alzheimer’s Disease is a major public health need. Keystone Bio have identified that specific virulent strains of Porphyromonas gingivalis (Pg) and the release of specific virulence factors/toxins in the oral cavity as the primary driver/causation of systemic/neuro‐vascular inflammation/degenerative diseases i.e., cognitive decline/dementias/Alzheimer’s disease (Nara et. al 2021). A recent landmark finding was reported in a sub‐cohort analysis from a Phase 2/3 GAIN trial (n = 238) demonstrated that lowering the load of Pg in the mouth leads to a significant improvement of cognitive slowing at both 24 and 48 weeks in participants with mild to moderate AD. Another study showed lowering the load of Pg in the mouth had a favorable effect on AD‐related brain atrophy (https://doi.org/10.1002/alz.12378). Pg OMVs and systemic system diseases has many well‐defined examples such as: cardiometabolic diseases‐ Pg OMVs attenuate insulin induced Akt/GSK‐3_ signaling in hepatic HepG2 cells, thereby causing changes in glucose metabolism in the liver and promoting the development of diabetes and increase vascular permeability by cleaving endothelial cell connexins such as PECAM‐1, thereby promoting cardiovascular diseases.Keystone Bio has further developed both a companion diagnostic (CDx) and a clinical, proof‐of‐concept tested, first generation, safe, efficacious, precision, bio‐therapeutic murine monoclonal antibody (KB‐001) for the diagnosis, treatment and monitoring against the oral bacteria and major virulent factor/toxin of Pg. The antibody engagement is with the later stages of the complex virulent factor/toxin secretion containing outer membrane vesicles from the bacteria thereby interfering/stopping all necessary metabolic, host defense, energy‐producing sources, adherence and biofilm formation and integrity (Nara et. al 2021). The talk will review what now seems like a solid case for causation in the role of virulent/toxin secreting strains of in neuro‐inflammation leading to cognitive decline, dementia, Sporadic Alzheimer’s disease and possibly Parkinson’s.

RNA is a key component of extracellular DNA networks in Pseudomonas aeruginosa biofilms

The extracellular matrix of bacterial biofilms consists of diverse components including polysaccharides, proteins and DNA. Extracellular RNA (eRNA) can also be present, contributing to the structural integrity of biofilms. However, technical difficulties related to the low stability of RNA make it difficult to understand the precise roles of eRNA in biofilms. Here, we show that eRNA associates with extracellular DNA (eDNA) to form matrix fibres in Pseudomonas aeruginosa biofilms, and the eRNA is enriched in certain bacterial RNA transcripts. Degradation of eRNA associated with eDNA led to a loss of eDNA fibres and biofilm viscoelasticity. Compared with planktonic and biofilm cells, the biofilm matrix was enriched in specific mRNA transcripts, including lasB (encoding elastase). The mRNA transcripts colocalised with eDNA fibres in the biofilm matrix, as shown by single molecule inexpensive FISH microscopy (smiFISH). The lasB mRNA was also observed in eDNA fibres in a clinical sputum sample positive for P. aeruginosa. Thus, our results indicate that the interaction of specific mRNAs with eDNA facilitates the formation of viscoelastic networks in the matrix of Pseudomonas aeruginosa biofilms.

Design, synthesis, and bioactivity studies of chalcone derivatives containing [1,2,4]-triazole-[4,3-a]-pyridine

In this study, 30 chalcone derivatives containing [1,2,4]-triazole-[4,3-a]-pyridine were designed and synthesized. The results of antibacterial activity showed that EC50 values of N26 against Xoo, Pcb was 36.41, 38.53 μg/mL, respectively, which were better than those of thiodiazole copper, whose EC50 values were 60.62, 106.75 μg/mL, respectively. The bacterial inhibitory activity of N26 against Xoo was verified by SEM. Antibacterial mechanism between N26 and Xoo was preliminarily explored, the experimental results showed that when the drug concentration was 100 mg/L, N26 had a good cell membrane permeability of Xoo, and it can inhibit the production of EPS content and extracellular enzyme content to disrupt the integrity of the Xoo biofilms achieving the effect of inhibiting Xoo. At 200 mg/L, N26 can protect and inhibit the lesions of post-harvested potatoes in vivo. The activities of N1-N30 against TMV were determined with half leaf dry spot method. The EC50 values of the curative and protective activity of N22 was 77.64 and 81.55 μg/mL, respectively, which were superior to those of NNM (294.27, 175.88 μg/mL, respectively). MST experiments demonstrated that N22 (Kd = 0.0076 ± 0.0007 μmol/L) had a stronger binding ability with TMV-CP, which was much higher than that of NNM (Kd = 0.7372 ± 0.2138 μmol/L). Molecular docking results showed that N22 had a significantly higher affinity with TMV-CP than NNM.