The Low-Abundance Plasma Proteome Reveals Differentially Abundant Proteins Associated with Breast Implant Capsular Contracture: A Pilot Study.

Capsular contracture (CC) is one of the most common postoperative complications associated with breast implant-associated infections. The mechanisms that lead to CC remain poorly understood. Plasma is an ideal biospecimen for early proteomics biomarker discovery. However, as high-abundance proteins mask signals from low-abundance proteins, identifying novel or specific proteins as biomarkers for a particular disease has been hampered. Here, we employed depletion of high-abundance plasma proteins followed by Tandem Mass Tag (TMT)-based quantitative proteomics to compare 10 healthy control patients against 10 breast implant CC patients. A total of 450 proteins were identified from these samples. Among them, 16 proteins were significantly differentially expressed in which 5 proteins were upregulated and 11 downregulated in breast implant CC patients compared to healthy controls. Gene Ontology enrichment analysis revealed that proteins related to cell, cellular processes and catalytic activity were highest in the cellular component, biological process, and molecular function categories, respectively. Further, pathway analysis revealed that inflammatory responses, focal adhesion, platelet activation, and complement and coagulation cascades were enriched pathways. The differentially abundant proteins from TMT-based quantitative proteomics have the potential to provide important information for future mechanistic studies and in the development of breast implant CC biomarkers.

Preclinical 3D-model supports an invisibility cloak for adenoid cystic carcinoma.

The tumour-cell based initiation of immune evasion project evaluated the role of Gipie in adenoid cystic carcinoma (ACC) and mucoepidermoid carcinoma (A-253), from ninety-six 3D-ACC and A-253-immune co-culture models using natural killer cells (NK), and Jurkat cells (JK). Abnormal ACC morphology was observed in 3D-ACC immune co-culture models. Gipie-silencing conferred a "lymphoblast-like" morphology to ACC cells, a six-fold increase in apoptotic cells (compared to unaltered ACC cells, P ≤ 0.0001), a two-fold decrease in T regulatory cells (FoxP3+/IL-2Rα+/CD25+) (P ≤ 0.0001), and a three-fold increase in activated NK cells (NKp30+/IFN-γ+) (P ≤ 0.0001) with significantly higher release of granzyme (P ≤ 0.001) and perforin (P ≤ 0.0001).

Topical Tranexamic Acid in Primary Breast Augmentation Surgery: Short- and Long-term Outcomes.

BACKGROUND: Breast augmentation mammaplasty (BAM) remains the most popular cosmetic procedure done worldwide. Bleeding in this procedure increases the chance of capsular contracture. Tranexamic acid (TXA), an antifibrinolytic, has been widely used by other surgical specialties to reduce bleeding. OBJECTIVES: We aimed to evaluate the use of TXA in BAM surgery. METHODS: This was a single-surgeon case series of all patients who underwent primary BAM from March 2017 to March 2018 and received topical TXA spray to the implant pocket before implant insertion. Early postoperative complications and long-term outcomes, such as capsular contracture and revisional surgery, were recorded and described. RESULTS: Two hundred and eighty-eight patients were included in the study with an overall complication rate of 2.8% over 5 years. No patients had postoperative bleeding or hematoma formation. One patient had a seroma, managed with ultrasound drainage. Complications requiring reoperation included rippling (3, 1.0%), pocket revision (2, 0.7%), capsule contracture (1, 0.3%) and rupture (1, 0.3%). CONCLUSIONS: This study highlights the safety and potential benefits of the use of topical TXA in breast augmentation, with low bleeding and capsular contracture rates.

Significant increased bacterial contamination with endoscope overnight and weekend storage times.

BACKGROUND AND AIM: Forced-air drying (FAD) cabinets are recommended for storage of reprocessed endoscopes, but financial constraints prevent their universal application. The study aimed to determine bacterial contamination in flexible gastroscopes (FG) channels after storage, in a cabinet with filtered air and UV lights, but without FAD. METHODS: Eight FG in clinical use in an endoscopy service of a large Brazilian hospital were sampled: immediately "Time zero" (N = 50), 12 h "Time 1" (N = 25), and 60 h "Time 2" (N = 25) after reprocessing. Following a flush-brush-flush of channels, 40-mL sterile water and 3 cm of the brush were collected. Each sample was divided, filtered onto two 0.22-µm membranes, and incubated in media without or with disinfectant neutralizer. Automated method was used for identification and antibiotic resistance test of isolated bacteria. RESULTS: Bacterial contamination in times "1" and "2" was 5.9 and 16.1 times greater than that of "Time zero," respectively. Number of positive cultures in media with and without neutralizer was similar at times "1" and "2," while media with neutralizer produced more positive cultures at "Time zero." Most bacteria isolated at "Time 2" were Gram-negative rods (52.3%) and showed resistance to one or more antibiotics (65%). CONCLUSION: Bacterial contamination was detected on reprocessed FG stored in non-FAD cabinets overnight (12 h) and increased with longer storage time (60 h). The contamination source is likely to be bacteria in biofilm which multiply in the absence of FAD. Evidence-based criteria should be available for storage time according to the cabinet available.

Staphylococcus aureus Cell Wall Phenotypic Changes Associated with Biofilm Maturation and Water Availability: A Key Contributing Factor for Chlorine Resistance.

Staphylococcus aureus biofilms are resistant to both antibiotics and disinfectants. As Staphylococci cell walls are an important defence mechanism, we sought to examine changes to the bacterial cell wall under different growth conditions. Cell walls of S. aureus grown as 3-day hydrated biofilm, 12-day hydrated biofilm, and 12-day dry surface biofilm (DSB) were compared to cell walls of planktonic organisms. Additionally, proteomic analysis using high-throughput tandem mass tag-based mass spectrometry was performed. Proteins involved in cell wall synthesis in biofilms were upregulated in comparison to planktonic growth. Bacterial cell wall width (measured by transmission electron microscopy) and peptidoglycan production (detected using a silkworm larva plasma system) increased with biofilm culture duration (p < 0.001) and dehydration (p = 0.002). Similarly, disinfectant tolerance was greatest in DSB, followed by 12-day hydrated biofilm and then 3-day biofilm, and it was least in the planktonic bacteria--suggesting that changes to the cell wall may be a key factor for S. aureus biofilm biocide resistance. Our findings shed light on possible new targets to combat biofilm-related infections and hospital dry surface biofilms.

Minimizing the Risks of Simethicone in Endoscope Reprocessing.

This statement was written under the auspices of the World Gastroenterology Organization's Guidelines Committee. The authors are members of the Review Team of the WGO Endoscope Disinfection Guideline and have experience in endoscopy, endoscope reprocessing, and microbiology, including biofilms. During the preparation of the WGO Update on Endoscope Disinfection Guidelines, concerns about simethicone on endoscope channel surfaces compromising cleaning and disinfection were raised. Publications on simethicone, including modes of delivery, effectiveness, and risks, have been reviewed. The paper was written as a companion to the new guidelines with a focus on minimizing the risks of simethicone in endoscope reprocessing.

Biofilme em parafusos ortopédicos prontos para uso adquiridos por meio de sistema de consiganação/comodato / Biopelícula en tornillos ortopédicos listos para uso adquiridos por medio del sistema de consignación/comodato / Biofilm on patient-ready orthopaedic screws acquired through the loaner system

Resumo Objetivo Avaliar a integridade da superfície e as condições microbiológicas de parafusos prontos para uso em bandejas ortopédicas após múltiplos processamentos. Métodos Após o processamento completo, as bandejas utilizadas em cirurgias de pequenos fragmentos, fornecidas por meio de sistema de consignação/comodato em um hospital brasileiro, foram selecionadas aleatoriamente durante quatro meses. Os parafusos mais utilizados (números 14, 16 e 18 - Grupo 1) e menos utilizados (números 10 e 38 - Grupo 2), portanto, os mais e menos expostos a agentes biológicos, químicos e físicos, foram aleatoriamente removidos e submetidos a inspeção visual (n=126), seguido de cultura bacteriana (n=6 parafusos/bandeja, 9 bandejas), teste de proteínas (n=6 parafusos/bandeja, 9 bandejas) e Microscopia Eletrônica de Varredura (MEV) (n=2 parafusos/bandeja, 9 bandejas). As culturas positivas foram submetidas a métodos automatizados de identificação bacteriana e suscetibilidade antimicrobiana. Resultados Foram detectadas ranhuras em 8,7% dos parafusos, predominantemente no Grupo 2 (8/11). Proteína residual foi detectada em 96,3%, e não houve diferença estatisticamente significativa na quantidade de proteína entre os grupos (P=0,07). Crescimento bacteriano foi identificado em 3/54 parafusos. Danos na superfície e presença de sujidade foram visualizados em todos os parafusos submetidos a MEV. Formação de biofilmes extensos foi detectada em oito parafusos, três do Grupo 1 e cinco do Grupo 2. Conclusão Recuperação de bactérias viáveis, acúmulo de biofilme e danos na superfície foram detectados nos parafusos prontos para uso. Os parafusos costumam permanecer nas bandejas cirúrgicas e serem submetidos a múltiplos processamento, sendo expostos a contaminação e danos repetidas vezes. Esses achados apontam para a necessidade de discutir e repensar a forma como esses implantes de uso único são atualmente disponibilizados para cirurgias.

Resumen Objetivo Evaluar la integridad de la superficie y las condiciones microbiológicas de tornillos listos para uso en bandejas ortopédicas después de múltiples procesamientos. Métodos Después del procesamiento completo, fueron seleccionadas aleatoriamente durante cuatro meses las bandejas utilizadas en cirugías de pequeños fragmentos, proporcionadas mediante el sistema de consignación/comodato en un hospital brasileño. Los tornillos más utilizados (números 14, 16 y 18 - Grupo 1) y menos utilizados (números 10 y 38 - Grupo 2), por lo tanto, los más y menos expuestos a agentes biológicos, químicos y físicos, fueron quitados aleatoriamente y sometidos a inspección visual (n=126), seguido de cultivo bacteriano (n=6 tornillos/bandeja, 9 bandejas), prueba de proteínas (n=6 tornillos/bandeja, 9 bandejas) y microscopía electrónica de barrido (MEB) (n=2 tornillos/bandeja, 9 bandejas). Los cultivos positivos fueron sometidos a métodos automatizados de identificación bacteriana y susceptibilidad antimicrobiana. Resultados Se detectaron ranuras en el 8,7 % de los tornillos, predominantemente en el Grupo 2 (8/11). Se detectó proteína residual en el 96,3 % y no se encontró diferencia estadísticamente significativa en la cantidad de proteína entre los grupos (P=0,07). En 3/54 tornillos se identificó crecimiento bacteriano. Se visualizaron daños en la superficie y presencia de suciedad en todos los tornillos sometidos a MEB. En ocho tornillos se detectó la formación de biopelículas, tres del Grupo 1 y cinco del Grupo 2. Conclusión Se detectó recuperación de bacterias viables, acumulación de biopelícula y daños en la superficie en los tornillos listos para uso. Los tornillos suelen permanecer en las bandejas quirúrgicas y son sometidos a múltiples procesamientos, donde están expuestos a contaminación y daños repetidas veces. Estos descubrimientos señalan la necesidad de discutir y repensar la forma como estos implantes de uso único se ponen a disposición para cirugía actualmente.

Abstract Objective Assess the surface integrity and microbiological conditions of patient-ready screws in orthopaedic trays that had been multiply reprocessed. Methods After full reprocessing, clinical trays used for small fragment surgery provided through a loaner system to a Brazilian hospital were randomly selected during four months. The most (numbers 14, 16 and 18 - Group 1) and least (numbers 10 and 38 - Group 2) frequently implanted screws, therefore, the ones estimated to be the most and least exposed to biological, chemical and physical agents, were randomly removed and subjected to visual inspection (n=126), followed by bacterial culture (n=6 screws/tray, 9 trays), protein test (n=6 screws/tray, 9 trays) and Scanning Electron Microscopy (SEM) (n=2 screws/tray, 9 trays). Positive cultures were subjected to automated bacterial identification and antimicrobial susceptibility tests. Results Grooves were detected on 8.7% screws, predominantly in Group 2 (8/11). Residual protein was detected on 96,3%, and there was no statistically significant difference in the amount of protein between the groups (P=0.07). Bacterial growth was identified in 3/54 screws. Surface damage and soil were visualized on all screws subjected to SEM. Extensive biofilms were detected on eight screws, three from Group 1 and five from Group 2. Conclusion Recovery of bacteria, biofilm accumulation and surface damage were detected on patient-ready screws. Screws frequently remain in surgical trays for multiple reprocessing; thus they are repeatedly exposed to contamination and damage. These findings point to the need to discuss and review the way these single-use implants are currently made available for surgeries.

One Step Forward with Dry Surface Biofilm (DSB) of Staphylococcus aureus: TMT-Based Quantitative Proteomic Analysis Reveals Proteomic Shifts between DSB and Hydrated Biofilm.

The Gram-positive bacterium Staphylococcus aureus is responsible for serious acute and chronic infections worldwide and is well-known for its biofilm formation ability. Recent findings of biofilms on dry hospital surfaces emphasise the failures in current cleaning practices and disinfection and the difficulty in removing these dry surface biofilms (DSBs). Many aspects of the formation of complex DSB biology on environmental surfaces in healthcare settings remains limited. In the present study, we aimed to determine how the protein component varied between DSBs and traditional hydrated biofilm. To do this, biofilms were grown in tryptic soy broth (TSB) on removable polycarbonate coupons in the CDC biofilm reactor over 12 days. Hydrated biofilm (50% TSB for 48 h, the media was then changed every 48 h with 20% TSB, at 37 °C with 130 rpm). DSB biofilm was produced in 5% TSB for 48 h at 35 °C followed by extended periods of dehydration (48, 66, 42 and 66 h at room temperature) interspersed with 6 h of 5% TSB at 35 °C. Then, we constructed a comprehensive reference map of 12-day DSB and 12-day hydrated biofilm associated proteins of S. aureus using a high-throughput tandem mass tag (TMT)-based mass spectrometry. Further pathway analysis of significantly differentially expressed identified proteins revealed that proteins significantly upregulated in 12-day DSB include PTS glucose transporter subunit IIBC (PtaA), UDP-N-acetylmuramate-L-alanine ligase (MurC) and UDP-N-acetylenolpyruvoylglucosamine (MurB) compared to 12-day hydrated biofilm. These three proteins are all linked with peptidoglycan biosynthesis pathway and are responsible for cell-wall formation and thicker EPS matrix deposition. Increased cell-wall formation may contribute to the persistence of DSB on dry surfaces. In contrast, proteins associated with energy metabolisms such as phosphoribosyl transferase (PyrR), glucosamine--fructose-6-phosphate aminotransferase (GlmS), galactose-6-phosphate isomerase (LacA), and argininosuccinate synthase (ArgG) were significantly upregulated whereas ribosomal and ABC transporters were significantly downregulated in the 12-day hydrated biofilm compared to DSB. However, validation by qPCR analysis showed that the levels of gene expression identified were only partially in line with our TMT-MS quantitation analysis. For the first time, a TMT-based proteomics study with DSB has shed novel insights and provided a basis for the identification and study of significant pathways vital for biofilm biology in this reference microorganism.

Mild Positive Pressure Improves the Efficacy of Benzalkonium Chloride against Staphylococcus aureus Biofilm.

Current protocols using liquid disinfectants to disinfect heat-sensitive hospital items frequently fail, as evidenced by the continued isolation of bacteria following decontamination. The contamination is, in part, due to biofilm formation. We hypothesize that mild positive pressure (PP) will disrupt this biofilm structure and improve liquid disinfectant/detergent penetration to biofilm bacteria for improved killing. Staphylococcus aureus biofilm, grown on polycarbonate coupons in the biofilm reactor under shear at 35 °C for 3 days, was treated for 10 min and 60 min with various dilutions of benzalkonium chloride without PP at 1 atmosphere (atm), and with PP at 3, 5, 7, and 10 atm. The effect on biofilm and residual bacterial viability was determined by standard plate counts, confocal laser scanning microscopy, and scanning electron microscopy. Combined use of benzalkonium chloride and PP up to 10 atm significantly increased biofilm killing up to 4.27 logs, as compared to the treatment using disinfectant alone. Microscopy results were consistent with the viability plate count results. PP improved disinfectant efficacy against bacterial biofilm. The use of mild PP is possible in many flow situations or if equipment/contaminated surfaces can be placed in a pressure chamber.

Proteome of Staphylococcus aureus Biofilm Changes Significantly with Aging.

Staphylococcus aureus is a notorious biofilm-producing pathogen that is frequently isolated from implantable medical device infections. As biofilm ages, it becomes more tolerant to antimicrobial treatment leading to treatment failure and necessitating the costly removal of infected devices. In this study, we performed in-solution digestion followed by TMT-based high-throughput mass spectrometry and investigated what changes occur in the proteome of S. aureus biofilm grown for 3-days and 12-days in comparison with 24 h planktonic. It showed that proteins associated with biosynthetic processes, ABC transporter pathway, virulence proteins, and shikimate kinase pathway were significantly upregulated in a 3-day biofilm, while proteins associated with sugar transporter, degradation, and stress response were downregulated. Interestingly, in a 3-day biofilm, we observed numerous proteins involved in the central metabolism pathways which could lead to biofilm growth under diverse environments by providing an alternative metabolic route to utilize energy. In 12-day biofilms, proteins associated with peptidoglycan biosynthesis, sugar transporters, and stress responses were upregulated, whereas proteins associated with ABC transporters, DNA replication, and adhesion proteins were downregulated. Gene Ontology analysis revealed that more proteins are involved in metabolic processes in 3dwb compared with 12dwb. Furthermore, we observed significant variations in the formation of biofilms resulting from changes in the level of metabolic activity in the different growth modes of biofilms that could be a significant factor in S. aureus biofilm maturation and persistence. Collectively, potential marker proteins were identified and further characterized to understand their exact role in S. aureus biofilm development, which may shed light on possible new therapeutic regimes in the treatment of biofilm-related implant-associated infections.

Management of surgical instruments at loaner companies in upper-middle and high-income countries: The other side of the coin.

BACKGROUND: The complex and specific surgical instruments (SI) acquired from loaner companies are provided non-sterile and must be reprocessed before each use. We evaluated the management practices of SI in loaner companies in a high (Australia) and a middle-income (Brazil) country. METHODS: Seven company managers in Australia and eight in Brazil replied to the self-administrated survey. RESULTS: Failures to meet recommended practices were detected, including standard operating procedure (SOP) provision, minimum delivery time, transport container and decontamination in loaner companies in both countries. Six of seven loaner companies in each country provided SOP for instrument reprocessing (one company in Brazil did not reply). Solicitation of the SI may occur 12 h before need in Brazil, and delivery of the set could be accepted 4 h before surgery in both countries. Transport of SI on stainless steel/aluminium trays was reported by 42.85% and 28.6% of companies in Brazil and Australia, respectively. In Australia, 57.1% of the loaner companies affirmed they performed cleaning only if the SI were visibly dirty. In Brazil, 62.5% of the companies reported they use alcohol to wipe SI. CONCLUSIONS: There is a need for standardisation of requirements and process verification improvement in loaner SI reprocessing and management.

Efficacy of Surgical/Wound Washes against Bacteria: Effect of Different In Vitro Models.

Topical antiseptics are often used to treat chronic wounds with biofilm infections and during salvage of biofilm contaminated implants, but their antibacterial efficacy is frequently only tested against non-aggregated planktonic or free-swimming organisms. This study evaluated the antibacterial and antibiofilm efficacy of four commercial surgical washes Bactisure, TorrenTX, minimally invasive lavage (MIS), and Betadine against six bacterial species: Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus pyogenes, Acinetobacter baumannii, Pseudomonas aeruginosa, and Escherichia coli, which are commonly isolated from surgical site infections and chronic wound infections using different in vitro models. We determined minimum planktonic inhibitory and eradication concentration and minimum 1-day-old biofilm inhibition and eradication concentration of antiseptics in 96-well plates format with 24 h contact time. We also tested the efficacy of antiseptics at in-use concentration and contact time in the presence of biological soil against 3-day-old biofilm grown on coupons with shear in a bioreactor, such that the results are more applicable to the clinical biofilm situations. In the 96-well plate model, the minimum concentration required to inhibit or kill planktonic and biofilm bacteria was lower for Bactisure and TorrenTX than for MIS and Betadine. However, Betadine and Bactisure showed better antibiofilm efficacy than TorrenTX and MIS in the 3-day-old biofilm bioreactor model at in-use concentration. The minimal concentration of surgical washes required to inhibit or kill planktonic bacterial cells and biofilms varies, suggesting the need for the development and use of biofilm-based assays to assess antimicrobial therapies, such as topical antiseptics and their effective concentrations. The antibiofilm efficacy of surgical washes against different bacterial species also varies, highlighting the importance of testing against various bacterial species to achieve a thorough understanding of their efficacy.

Biofilm on Toothbrushes of Children with Cystic Fibrosis: A Potential Source of Lung Re-Infection after Antibiotic Treatment?

Frequent recurrent lung infections result in irreversible lung damage in children with cystic fibrosis (CF). This study aimed to determine if toothbrushes contain biofilms of pathogens, and act as potential reservoirs for lung re-infection following antibiotic treatment of acute exacerbations. Toothbrushes were collected from children with CF of lung infection before, during and after antibiotic treatment. Toothbrushes were rinsed with sterile saline and cultured. Bacterial isolates from toothbrushes were identified by 16s rRNA gene sequencing and compared with isolates from a sputum sample of the same patient. Scanning electron microscopy (SEM) was used to visually confirm the presence of bacterial biofilms and confocal laser scanning microscopy (CLSM) combined with Live/Dead stain to confirm bacterial viability. Large numbers of bacteria and biofilms were present on all toothbrushes. SEM confirmed the presence of biofilms and CLSM confirmed bacterial viability on all toothbrushes. Pathogens identified on toothbrushes from children before and during antibiotics treatment were in concordance with the species found in sputum samples. Pseudomonas aeruginosa and Staphylococcus aureus was able to be cultured from children's toothbrushes despite antibiotic treatment. Toothbrushes were shown to be contaminated with viable pathogens and biofilms before and during antibiotic treatment and could be a potential source of lung re-infections.

Reprocessing of loaned surgical instruments/implants in Australia and Brazil: A survey of those at the coalface.

BACKGROUND: Acquisition of surgical instruments (SI) and implants by loaner system is a worldwide practice. Although quality of device reprocessing has been reported to be lower in low and middle-income countries compared with higher income countries, few investigations have been conducted on the management and reprocessing of loaned devices. Thus, in this study we evaluate the practices of management and reprocessing of loaner SI and non-sterile implants (NSI) in a high (Australia) and a middle-income (Brazil) country. METHODS: Cross-sectional surveys (hardcopy or online form) was self-administered by sterilising service unit's personnel of Brazilian and Australian hospitals. Only those units that sourced orthopaedic surgical instruments and non-sterile loaned implants from independent companies were eligible. RESULTS: A total of 65 questionnaires from Australia and 168 from Brazil were analysed. Quality indicators regarding structure and work process for the management and reprocessing of loaned SI and NSI was of a higher standard in Australia than in Brazil. However, failures were detected in both countries, for instance delivery delays and improper point-of-use pre-cleaning practices. CONCLUSION: There are key challenges inherent of the multifaceted reusable medical devices loaner system in both countries, such as communication failures, implementation of non-recommended practices, reprocessing of NSI, and other related to structure and process quality indicators, that must be faced. Initial and ongoing education and training should be provided and should embrace the themes of technical proficiency, effective communication and teamwork, and should include all personnel involved in this process, even loaner company staff.

Biofilm accumulation in new flexible gastroscope channels in clinical use.

OBJECTIVE: Assess the accumulation of protein and biofilm on the inner surfaces of new flexible gastroscope (FG) channels after 30 and 60 days of patient use and full reprocessing. DESIGN: Clinical use study of biofilm accumulation in FG channels. SETTING: Endoscopy service of a public hospital. METHODS: First, we tested an FG in clinical use before the implementation of a revised reprocessing protocol (phase 1 baseline; n = 1). After replacement of the channels by new ones and the implementation of the protocol, 3 FGs were tested after 30 days of clinical use (phase 2; n = 3) and 3 FGs were tested after 60 days of clinical use (phase 3; n = 3), and the same FGs were tested in phase 2 and 3. Their biopsy, air, water, and air/water junction channels were removed and subjected to protein testing (n = 21), bacteriological culture (n = 21), and scanning electron microscopy (SEM) (n = 28). Air-water junction channels fragments were subjected to SEM only. RESULTS: For the FGs, the average number of uses and reprocessing cycles was 60 times. Extensive biofilm was detected in air, water, and air-water junction channels (n = 18 of 28). All channels (28 of 28) showed residual matter, and structural damage was identified in most of them (20 of 28). Residual protein was detected in the air and water channels of all FG evaluated (phases 1-3), except for 1 air channel from phase 2. Bacteria were recovered from 8 of 21 channels, most air or water channels. CONCLUSIONS: The short time before damage and biofilm accumulation in the channels was evident and suggests that improving the endoscope design is necessary. Better reprocessing methods and channel maintenance are needed.

Gram-Negative Bacterial Lipopolysaccharide Promotes Tumor Cell Proliferation in Breast Implant-Associated Anaplastic Large-Cell Lymphoma.

Breast implant-associated anaplastic large-cell lymphoma (BIA-ALCL) is a distinct malignancy associated with textured breast implants. We investigated whether bacteria could trigger the activation and multiplication of BIA-ALCL cells in vitro. BIA-ALCL patient-derived BIA-ALCL tumor cells, BIA-ALCL cell lines, cutaneous ALCL cell lines, an immortal T-cell line (MT-4), and peripheral blood mononuclear cells (PBMC) from BIA-ALCL, capsular contracture, and primary augmentation patients were studied. Cells were subjected to various mitogenic stimulation assays including plant phytohemagglutinin (PHA), Gram-negative bacterial lipopolysaccharide (LPS), Staphylococcal superantigens enterotoxin A (SEA), toxic shock syndrome toxin-1 (TSST-1), or sterilized implant shells. Patient-derived BIA-ALCL tumor cells and BIA-ALCL cell lines showed a unique response to LPS stimulation. This response was dampened significantly in the presence of a Toll-like receptor 4 (TLR4) inhibitor peptide. In contrast, cutaneous ALCL cells, MT-4, and PBMC cells from all patients responded significantly more to PHA, SEA, and TSST-1 than to LPS. Breast implant shells of all surface grades alone did not produce a proliferative response of BIA-ALCL cells, indicating the breast implant does not act as a pro-inflammatory stimulant. These findings indicate a possible novel pathway for LPS to promote BIA-ALCL cell proliferation via a TLR4 receptor-mediated bacterial transformation of T-cells into malignancy.

ML218 HCl Is More Efficient Than Capsaicin in Inhibiting Bacterial Antigen-Induced Cal 27 Oral Cancer Cell Proliferation.

The bacterial antigen, lipopolysaccharide (LPS) and disruptions in calcium channels are independently known to influence oral cancer progression. Previously, we found that bacterial antigens, LPS and lipoteichoic acid (LTA) act as confounders during the action of capsaicin on Cal 27 oral cancer proliferation. As calcium channel drugs may affect oral cancer cell proliferation, we investigated the effect of ML218 HCl, a T-type voltage-gated calcium channel blocker, on the proliferation of Cal 27 oral cancer cells. We hypothesized that ML218 HCl could effectively reduce LPS-induced oral cancer cell proliferation. LPS and LTA antigens were added to Cal 27 oral cancer cells either prior to and/or concurrently with ML218 HCl treatment, and the efficacy of the treatment was evaluated by measuring Cal 27 proliferation, cell death and apoptosis. ML218 HCl inhibited oral cancer cell proliferation, increased apoptosis and cell death, but their efficacy was significantly reduced in the presence of bacterial antigens. ML218 HCl proved more effective than capsaicin in reducing bacterial antigen-induced Cal 27 oral cancer cell proliferation. Our results also suggest an interplay of proliferation factors during the bacterial antigens and calcium channel drug interaction in Cal 27. Bacterial antigen reduction of drug efficacy should be considered for developing newer pharmacological agents or testing the efficacy of the existing oral cancer chemotherapeutic agents. Finally, voltage gated calcium channel drugs should be considered for future oral cancer research.

Bacterial Antigens Reduced the Inhibition Effect of Capsaicin on Cal 27 Oral Cancer Cell Proliferation.

Oral cancer is a major global health problem with high incidence and low survival rates. The oral cavity contains biofilms as dental plaques that harbour both Gram-negative and Gram-positive bacterial antigens, lipopolysaccharide (LPS) and lipoteichoic acid (LTA), respectively. LPS and LTA are known to stimulate cancer cell growth, and the bioactive phytochemical capsaicin has been reported to reverse this effect. Here, we tested the efficacy of oral cancer chemotherapy treatment with capsaicin in the presence of LPS, LTA or the combination of both antigens. LPS and LTA were administered to Cal 27 oral cancer cells prior to and/or concurrently with capsaicin, and the treatment efficacy was evaluated by measuring cell proliferation and apoptotic cell death. We found that while capsaicin inhibits oral cancer cell proliferation and metabolism (MT Glo assay) and increases cell death (Trypan blue exclusion assay and Caspase 3/7 expression), its anti-cancer effect was significantly reduced on cells that are either primed or exposed to the bacterial antigens. Capsaicin treatment significantly increased oral cancer cells' suppressor of cytokine signalling 3 gene expression. This increase was reversed in the presence of bacterial antigens during treatment. Our data establish a rationale for clinical consideration of bacterial antigens that may interfere with the treatment efficacy of oral cancer.

A multiomics approach to identify host-microbe alterations associated with infection severity in diabetic foot infections: a pilot study.

Diabetic foot infections (DFIs) are a major cause of hospitalization and can lead to lower extremity amputation. In this pilot study, we used a multiomics approach to explore the host-microbe complex within DFIs. We observed minimal differences in the overall microbial composition between PEDIS infection severities, however Staphylococcus aureus and Streptococcus genera were abundant and highly active in most mild to moderate DFIs. Further, we identified the significant enrichment of several virulence factors associated with infection pathogenicity belonging to both Staphylococcus aureus and Streptococcus. In severe DFIs, patients demonstrated a greater microbial diversity and differential gene expression demonstrated the enrichment of multispecies virulence genes suggestive of a complex polymicrobial infection. The host response in patients with severe DFIs was also significantly different as compared to mild to moderate DFIs. This was attributed to the enrichment of host genes associated with inflammation, acute phase response, cell stress and broad immune-related responses, while those associated with wound healing and myogenesis were significantly depleted.