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Cold atmospheric plasmas (CAPs) within recent years have shown great promise in the field of plasma medicine, encompassing a variety of treatments from wound healing to the treatment of cancerous tumors. For each subsequent treatment, a different application of CAPs has been postulated and attempted to best treat the target for the most effective results. These treatments have varied through the implementation of control parameters such as applied settings, electrode geometries, gas flow, and the duration of the treatment. However, with such an extensive number of variables to consider, scientists and engineers have sought a means to accurately control CAPs for the best-desired effects in medical applications. This paper seeks to investigate and characterize the historical precedent for the use of plasma control mechanisms within the field of plasma medicine. Current control strategies, plasma parameters, and control schemes will be extrapolated through recent developments and successes to gain better insight into the future of the field and the challenges that are still present in the overall implementation of such devices. Proposed approaches, such as data-driven machine learning, and the use of closed-loop feedback controls, will be showcased as the next steps toward application.

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Introduction. Aspergillus flavus and Fusarium keratoplasticum are common causative pathogens of fungal keratitis (FK), a severe corneal disease associated with significant morbidity and vision loss. Escalating incidence of antifungal resistance to available antifungal drugs poses a major challenge to FK treatment. Cold atmospheric plasma (CAP) is a pioneering nonpharmacologic antimicrobial intervention that has demonstrated potential as a broad-spectrum antifungal treatment.Gap statement. Previous research highlights biofilm-associated resistance as a critical barrier to effective FK treatment. Although CAP has shown promise against various fungal infections, its efficacy against biofilm and conidial forms of FK pathogens remains inadequately explored.Aim. This study aims to investigate the antifungal efficacy of CAP against clinical fungal keratitis isolates of A. flavus and F. keratoplasticum in vitro.Methodology. Power parameters (22-27 kVpp, 300-400 Hz and 20-80 mA) of a dielectric barrier discharge CAP device were optimized for inactivation of A. flavus biofilms. Optimal applied voltage and total current were applied to F. keratoplasticum biofilms and conidial suspensions of A. flavus and F. keratoplasticum. The antifungal effect of CAP treatment was investigated by evaluating fungal viability through means of metabolic activity, c.f.u. enumeration (c.f.u. ml-1) and biofilm formation.Results. For both fungal species, CAP exhibited strong time-dependent inactivation, achieving greater than 80 % reduction in metabolic activity and c.f.u. ml-1 within 300 s or less, and complete inhibition after 600 s of treatment.Conclusion. Our findings indicate that CAP is a promising broad-spectrum antifungal intervention. CAP treatment effectively reduces fungal viability in both biofilm and conidial suspension cultures of A. flavus and F. keratoplasticum, suggesting its potential as an alternative treatment strategy for fungal keratitis.

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INTRODUCTION: The renin-angiotensin system (RAS) has been demonstrated to modulate cell proliferation, desmoplasia, angiogenesis and immunosuppression. We examined the association of RAS inhibitors (RASi)-namely angiotensin converting enzyme inhibitors (ACEi) and angiotensin receptor blockers (ARB)-with neoadjuvant chemotherapy (NAC) for muscle-invasive bladder cancer (MIBC) preceding radical cystectomy (RC). PATIENTS AND METHODS: We retrospectively investigated concurrent RASi use with NAC prior to RC in 302 patients with MIBC from 3 academic institutions. Outcomes included pathologic complete response (pCR) and overall survival (OS). Pathologic features, performance status (PS), clinical stage, type/number of cycles of NAC, and toxicities were collected. RESULTS: Overall pCR rate was 26.2% and 5-year OS was 62%. Concurrent ACEi intake with NAC approached significance for association with pCR (odds ratio [OR] = 1.71; 95% CI, 0.94-3.11; P = .077). Patients with cT3/4N0-N1 disease receiving ACEi had higher pCR rates (30.8% vs. 17.7%, P = .056) than those not on ACEi. Female sex had a statistically significant favorable interaction for pCR with ACEi intake (P = .044). ACEi intake was not associated with OS, while pCR, PS and lower clinical stage were significantly associated with improved OS. CONCLUSION: ACEi intake is potentially associated with increased pCR in patients with MIBC receiving NAC prior to RC, and this association is more pronounced in patients with higher clinical stage of disease at the initiation of therapy and female sex. Our data suggest the potential relevance of the RAS as a therapeutic target in aggressive MIBC.

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BACKGROUND CONTEXT: The influence of SDOH on spine surgery is poorly understood. Historically, researchers commonly focused on the isolated influences of race, insurance status, or income on healthcare outcomes. However, analysis of SDOH is becoming increasingly more nuanced as viewing social factors in aggregate rather than individually may offer more precise estimates of the impact of SDOH on healthcare delivery. PURPOSE: The aim of this study was to evaluate the effects of patient social history on length of stay (LOS) and readmission within 90 days following spine surgery using ensemble machine learning and multilayer perceptron. STUDY DESIGN: Retrospective chart review. PATIENT SAMPLE: 8,565 elective and emergency spine surgery cases performed from 2013 to 2023 using our institution's database of longitudinally collected electronic medical record information. OUTCOMES MEASURES: Patient LOS, discharge disposition, and rate of 90-day readmission. METHODS: Ensemble machine learning and multilayer perceptron were employed to predict LOS and readmission within 90 days following spine surgery. All other subsequent statistical analysis was performed using SPSS version 28. To further assess correlations among variables, Pearson's correlation tests and multivariate linear regression models were constructed. Independent sample t-tests, paired sample t-tests, one-way analysis of variance (ANOVA) with post-hoc Bonferroni and Tukey corrections, and Pearson's chi-squared test were applied where appropriate for analysis of continuous and categorical variables. RESULTS: Black patients demonstrated a greater LOS compared to white patients, but race and ethnicity were not significantly associated with 90-day readmission rates. Insured patients had a shorter LOS and lower readmission rates compared to non-insured patients, as did privately insured patients compared to publicly insured patients. Patients discharged home had lower LOS and lower readmission rates, compared to patients discharged to other facilities. Marriage decreased both LOS and readmission rates, underweight patients showcased increased LOS and readmission rates, and religion was shown to impact LOS and readmission rates. When utilizing patient social history, lab values, and medical history, machine learning determined the top 5 most-important variables for prediction of LOS -along with their respective feature importances-to be insurance status (0.166), religion (0.100), ICU status (0.093), antibiotic use (0.061), and case status: elective or urgent (0.055). The top 5 most-important variables for prediction of 90-day readmission-along with their respective feature importances-were insurance status (0.177), religion (0.123), discharge location (0.096), emergency case status (0.064), and history of diabetes (0.041). CONCLUSIONS: This study highlights that SDOH is influential in determining patient length of stay, discharge disposition, and likelihood of readmission following spine surgery. Machine learning was utilized to accurately predict LOS and 90-day readmission with patient medical history, lab values, and social history, as well as social history alone.

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BACKGROUND: Nonsteroidal anti-inflammatory drug (NSAID) use has been investigated as a modifiable risk factor for postoperative pancreatic fistula (POPF) after pancreatoduodenectomy (PD). This study comprises a systematic review and meta-analysis examining the impact of perioperative NSAID use on rates of POPF after PD. METHODS: A Preferred Reporting Items for Systematic Reviews and Meta-Analyses 2020-compliant systematic review was performed. Pooled mean differences (MD), odds ratios (OR), and risk ratios with 95% CIs were calculated. RESULTS: Seven studies published from 2015 to 2021 were included, reporting 2851 PDs (1372 receiving NSAIDs and 1479 not receiving NSAIDs). There were no differences regarding blood loss (MD -99.40 mL; 95% CI, -201.71 to 2.91; P = .06), overall morbidity (OR 1.05; 95% CI, 0.68-1.61; P = .83), hemorrhage (OR 2.35; 95% CI, 0.48-11.59; P = .29), delayed gastric emptying (OR 0.98; 95% CI, 0.60-1.60; P = .93), bile leak (OR 0.68; 95% CI, 0.12-3.89; P = .66), surgical site infection (OR 1.02; 95% CI, 0.33-3.22; P = .97), abscess (OR 0.99; 95% CI, 0.51-1.91; P = .97), clinically relevant POPF (OR 1.18; 95% CI, 0.84-1.64; P = .33), readmission (OR 0.94; 95% CI, 0.61-1.46; P = .78), or reoperation (OR 0.82; 95% CI, 0.33-2.06; P = .68). NSAID use was associated with a shorter hospital stay (MD -1.05 days; 95% CI, -1.39 to 0.71; P < .00001). CONCLUSION: The use of NSAIDs in the perioperative period for patients undergoing PD was not associated with increased rates of POPF.

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AIMS: Urinary tract infections are the most common hospital-acquired infection, 80% of which are associated with catheterization. Diagnostic methods may influence the reported identities of these pathogens, and phenotypic testing under laboratory conditions may not reflect infection phenotypes. This study aimed to evaluate the efficacy of diagnostic methods and whether medium composition alters phenotypes by characterizing catheter-associated urinary tract infection isolates from a UK hospital. METHODS AND RESULTS: We compared five bacterial identification methods, including biochemical testing, matrix-assisted laser desorption/ionization biotyping, and genome sequencing, finding differences in genus- or species-level identifications. Antibiotic susceptibility comparisons between phenotypic assays and genomic predictions showed high agreement only in multidrug-resistant strains. To determine whether growth rate and biofilm formation were affected by medium composition, strains were grown in both planktonic and biofilm states. Low planktonic growth and significant biofilm formation were observed in artificial urine compared to rich laboratory media, underscoring the importance of assay design. CONCLUSIONS: This study highlights the risks of relying on a single diagnostic method for species identification, advocating for whole-genome sequencing for accuracy. It emphasizes the continued importance of phenotypic methods in understanding antibiotic resistance in clinical settings and the need for characterization conditions that mirror those encountered by pathogens in the body.

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Stevens Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are severe cutaneous adverse reactions characterised by keratinocyte apoptosis, necroptosis and epidermal detachment. Several cytokines and cytotoxic proteins have been shown to be elevated in the blood and skin of SJS/TEN sufferers and biologics such as intravenous immune globulin and tumour necrosis factor (TNF)-alpha inhibitors have demonstrated good therapeutic potential. The exact pathogenic model of SJS/TEN however remains elusive. This systematic review aimed to evaluate the case-control studies of cytokines and cytotoxic proteins in the blister fluid and skin of adults with Stevens Johnson syndrome and/or toxic epidermal necrolysis. This review was registered with INPLASY and conducted in accordance with the PRISMA reporting guidelines. Potential bias was assessed using the NIH criteria. Eleven articles describing results from 96 cases and 170 controls were included. Fas, Fas ligand, Interleukin (IL)-8 and B-cell lymphoma (Bcl)-2 were elevated in SJS/TEN blister fluid and skin tissue, compared with healthy controls. IL-2, IL-6, TNF-alpha, tumour necrosis factor-related apoptosis-inducing ligand (TRAIL), interferon-gamma and matrix metalloproteinase-2 were elevated in SJS/TEN blister fluid compared with fluid sampled from lesional controls. Granulysin, IL-33, TGF-beta-1 and IL-13 were elevated in SJS/TEN skin tissue compared with lesional lichen planus tissue, as was IL-13, IFN-gamma, IL-2 and IL-5, when compared with erythema multiforme tissue. A wide array of cytokines and cytotoxic proteins are present at higher concentrations in the blister fluid and skin tissue of SJS/TEN patients compared with healthy and lesional controls. Our findings suggest that these proteins may be pathogenic, as well as possibly markers for diagnosis, disease severity and course. They may also prove to be useful therapeutic targets. More research is needed.

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Objectives This is the protocol for a Campbell systematic review. The objectives are as follows: The aim of this systematic review is to examine the scientific evidence available from low- and middle-income countries on the association of breakfast consumption habits and anthropometry/adiposity- and nutrition-related outcomes in adolescents aged 10-19 years old.

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Importance: Telehealth has the potential to improve health for older adults, but many access disparities exist, including for those with vision impairment (VI). Objective: To examine the associations between VI and digital technology access measures in US older adults. Design, Setting, and Participants: This was a cross-sectional study that included Medicare beneficiaries 65 years and older. Beneficiary data were obtained from the National Health and Aging Trends Study (NHATS) 2021; data collection was conducted from June 2020 through January 2021. Data were analyzed September 2023. Exposure: VI, measured on categorical (distance and near VI [>0.30 logMAR], contrast sensitivity impairment [CSI; <1.55 logCS], and any VI [distance VI, near VI, or CSI]), and continuous (distance and near acuity [logMAR] and contrast sensitivity [logCS]) scales. Main Outcomes: Self-reported outcomes of digital technology access from the technological environment component of the NHATS. Results: Of the 2822 Medicare beneficiaries (mean [SD] age, 78.5 [5.6] years; 1605 female [54.7%]) included in this study, patients self-identified with the following race and ethnicity categories (weighted percentages): 575 non-Hispanic Black (8.0%), 132 Hispanic (7.0%), 2019 non-Hispanic White (81.7%), and 63 non-Hispanic other race (3.4%), which included American Indian, Alaska Native, Asian, multiracial, Native Hawaiian, Pacific Islander, or other specified race. A total of 1077 of 2822 patients (32.3%) had any VI. In multivariable logistic regression models, older adults with any VI had lower odds of having or knowing how to use a cellphone (odds ratio [OR], 0.58; 95% CI, 0.38-0.88), computer (OR, 0.61; 95% CI, 0.47-0.79), or tablet (OR, 0.68, 95% CI = 0.54-0.85) than peers without VI. In other models, near VI was associated with lower odds of having and knowing how to use a phone (OR, 0.56; 95% CI, 0.36-0.87), computer (OR, 0.57; 95% CI, 0.44-0.75), or tablet (OR, 0.65; 95% CI, 0.52-0.81) compared with no near VI. CSI was associated with lower odds of having and knowing how to use a phone (OR, 0.66; 95% CI, 0.45-0.99) or computer (OR, 0.72; 95% CI, 0.55-0.93) compared with no CSI. Distance VI was only associated with lower odds of having and knowing how to use a cellphone (OR, 0.63; 95% CI, 0.41-0.95). Any VI and individual categorical VI measures were not associated with other outcomes of digital health- and nonhealth-related experiences. Similar associations were noted when vision was examined on a continuous scale. However, worse distance acuity (per 0.1 logMAR) was associated with being less likely to visit with family or friends on a video call (OR, 0.91; 95% CI, 0.84-0.98) and to order or refill prescriptions online (OR, 0.90; 95% CI, 0.83-0.97). Conclusions and Relevance: Results suggest that older US adults with VI were less likely to have access to digital technology than peers without VI, although no differences were noted in the report of digital health- and nonhealth-related activities. These findings highlight the potential for inequities that may arise in telehealth for older adults with VI and the necessity to develop strategies to improve accessibility of telemedicine for all.

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INTRODUCTION: Parkinson's disease (PKD) is neurodegenerative disorder marked by tremors, bradykinesia, muscle rigidity and reduction in precise hand movements which could lead to improper oral hygiene and Periodontal disease. Current systematic review aims to review existing literature and provide assessment of periodontal health in PKD patients through a meta-analysis METHODS: Review was performed in accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines and registered in PROSPERO-CRD42023451700. Databases were searched for studies having sufficient data on periodontal health in patients with PKD compared to healthy controls and reporting outcomes in terms of various periodontal parameters of probing depth (PD), plaque index (PI), clinical attachment level (CAL), presence of bleeding on probing and gingivitis. Quality assessment of included was evaluated using Newcastle Ottawa Scale (NOS). RESULTS: Eleven studies fulfilled the eligibility criteria, of which ten studies were suitable for meta-analysis. Pooled estimate through the SMD showed that all periodontal parameters were altered and significantly deteriorated in PKD compared to controls but this group differences were statistically insignificant (p > 0.05). Publication bias through the funnel plot showed symmetric distribution with absence of systematic heterogeneity. CONCLUSION: Parkinson's disease could be a possible factor in deterioration of periodontal health.

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BACKGROUND: Comprehensive profiling of autoantibodies (AAbs) in metastatic urothelial cancer (mUC) has not been performed to date. This may aid in diagnosis of UC, uncover novel therapeutic targets in this disease as well as identify associations between AAbs and response and toxicity to systemic therapies. METHODS: We used serum from patients with mUC collected prior to and after systemic therapy (immune checkpoint inhibitor (ICI) or platinum-based chemotherapy (PBC)) at Dana-Farber Cancer Institute. 38 age-matched and sex-matched healthy controls (HCs) from healthy blood donors were also evaluated. The SeroTag immuno-oncology discovery array (Oncimmune) was used, with quantification of the AAb reactivity toward 1132 antigens. Bound AAbs were detected using an anti-immunoglobulin G-specific detection antibody conjugated to the fluorescent reporter dye phycoerythrin. The AAb reactivity was reported as the median fluorescence intensity for each color and sample using a Luminex FlexMAP3D analyzer. Clinical outcomes of interest included radiographic response and development of immune-related adverse events (irAEs). Significance analysis of microarray was used to compare mUC versus HC and radiographic response. Associations with irAE were evaluated using a logistic regression model. P<0.05 was considered statistically significant. RESULTS: 66 patients were included with a median age of 68 years; 54 patients (82%) received ICI and 12 patients (18%) received PBC. Compared with HCs, AAbs against the cancer/testis antigens (CTAG1B, CTAG2, MAGEB18), HSPA1A, TP53, KRAS, and FGFR3 were significantly elevated in patients with mUC. AAbs against BRCA2, TP53, and CTNBB1 were associated with response, and those against BICD2 and UACA were associated with resistance to ICI therapy. AAbs against MITF, CDH3, and KDM4A were associated with development of irAEs in patient who received an ICI. A higher variance in pre-to-post treatment fold change in AAb levels was seen in patients treated with ICI versus PBC and was associated with response to ICI. CONCLUSIONS: This is the first report of comprehensive AAb profiling of patients with mUC and identified key AAbs that were elevated in patients with mUC versus HCs as well as AAbs associated with therapeutic response to ICI. These findings are hypothesis generating and further mechanistic studies evaluating humoral immunity in UC are required.

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PURPOSE: This study aimed to investigate the potential link between methamphetamine use and infectious syphilitic uveitis through a retrospective chart review of four patients with both conditions. METHODS: A retrospective chart review was conducted on four patients with a prior history of methamphetamine use and presented with vision loss at an emergency room or clinic that was ultimately diagnosed as syphilitic uveitis. RESULTS: The average age of the four patients was 41.5 ± 9.8 years old, and each presented with symptoms of blurry vision after an average of 3 weeks. Clinical and serologic examinations revealed variable severity of syphilitic uveitis in each patient, and a confirmed history of methamphetamine use was found on a review of social history. Each patient had worsening visual outcomes after the initial presentation and diagnosis. CONCLUSION: Syphilitic uveitis is a challenging diagnosis due to its similarity with other ocular pathologies. This case series highlights the importance of maintaining a high level of suspicion for syphilis in patients with a history of methamphetamine use and uveitis to enable rapid treatment and prevent visual decline. The poor visual outcomes observed in these four patients underscore the significance of identifying and treating syphilitic uveitis promptly in this population and the barriers to treatment completion.

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A woman in her 50s presented with suspected open globe injury (OGI) of the right eye after being hit with a high velocity piece of plastic. Visual acuity at the time of presentation was counting fingers in the affected eye. Slit lamp examination revealed a full thickness laceration of the cornea and a traumatic cataract. Primary corneal repair was performed and the patient was left aphakic after cataract removal. Secondary intraocular lens placement was deferred for 2 years, after which time a scleral-fixated intraocular lens was implanted using the Yamane technique. Postoperative visual acuity of 20/50 was achieved, with the vision limited by persistent diabetic macular oedema. Thus, this case of successful implantation of a secondary lens using the Yamane technique in a patient with prior corneal laceration and traumatic cataract highlights that the Yamane technique can result in visual improvement in patients with prior OGI.

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Catheter-associated urinary tract infections (CAUTIs) represent one of the major healthcare-associated infections, and Pseudomonas aeruginosa is a common Gram-negative bacterium associated with catheter infections in Egyptian clinical settings. The present study describes the phenotypic and genotypic characteristics of 31 P. aeruginosa isolates recovered from CAUTIs in an Egyptian hospital over a 3 month period. Genomes of isolates were of good quality and were confirmed to be P. aeruginosa by comparison to the type strain (average nucleotide identity, phylogenetic analysis). Clonal diversity among the isolates was determined; eight different sequence types were found (STs 244, 357, 381, 621, 773, 1430, 1667 and 3765), of which ST357 and ST773 are considered to be high-risk clones. Antimicrobial resistance (AMR) testing according to European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines showed that the isolates were highly resistant to quinolones [ciprofloxacin (12/31, 38.7 %) and levofloxacin (9/31, 29 %) followed by tobramycin (10/31, 32.5 %)] and cephalosporins (7/31, 22.5 %). Genotypic analysis of resistance determinants predicted all isolates to encode a range of AMR genes, including those conferring resistance to aminoglycosides, ß-lactamases, fluoroquinolones, fosfomycin, sulfonamides, tetracyclines and chloramphenicol. One isolate was found to carry a 422 938 bp pBT2436-like megaplasmid encoding OXA-520, the first report from Egypt of this emerging family of clinically important mobile genetic elements. All isolates were able to form biofilms and were predicted to encode virulence genes associated with adherence, antimicrobial activity, anti-phagocytosis, phospholipase enzymes, iron uptake, proteases, secretion systems and toxins. The present study shows how phenotypic analysis alongside genomic analysis may help us understand the AMR and virulence profiles of P. aeruginosa contributing to CAUTIs in Egypt.

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Purpose: To compare the lobbying expenditures and political action committee (PAC) campaign finance activities of the American Academy of Ophthalmology (AAO), American Society of Cataract and Refractive Surgery (ASCRS), and American Optometric Association (AOA) from 2015 to 2022. Methods: Financial data were collected from the Federal Election Commission and OpenSecrets database. Analysis was performed to characterize and compare financial activity among the organizations. P < 0.05 was considered significant and all analyses were two-sided. Results: From 2015 to 2022, the AAO, ASCRS, and AOA spent $6,745,000, $5,354,406, and $13,335,000 on lobbying, respectively. The AOA's annual lobbying expenditure (median, $1,725,000) was significantly greater than AAO's ($842,500, P = 0.03) and ASCRS's ($694,289, P < 0.001). In PAC donations, OPHTHPAC, affiliated with AAO, received $3,221,737 from 2079 donors (median, $900); eyePAC, affiliated with ASCRS, received $506,255 from 349 donors ($500); and AOA-PAC received $6,642,588 from 3641 donors ($825). Compared to eyePAC, median donations to OPHTHPAC (P = 0.01) and AOA-PAC (P = 0.04) were significantly higher. In campaign spending, OPHTHPAC contributed $2,728,500 to 326 campaigns (median, $5000), eyePAC contributed $293,500 to 58 campaigns ($3000), and AOA-PAC contributed $5,128,673 to 617 campaigns ($5500). eyePAC's median campaign contribution was significantly lower than the AOA's (P < 0.001) and AAO's (P = 0.007). Every PAC directed most of its contributions toward Republican campaigns; eyePAC donated the highest proportion (64.9%). Conclusions: AOA was more assertive in shaping policy by increasing lobbying expenditures, fundraising, and donating to a greater number of election campaigns.

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Histone H2A lysine 119 (H2AK119Ub) is monoubiquitinated by Polycomb repressive complex 1 and deubiquitinated by Polycomb repressive deubiquitinase complex (PR-DUB). PR-DUB cleaves H2AK119Ub to restrict focal H2AK119Ub at Polycomb target sites and to protect active genes from aberrant silencing. The PR-DUB subunits (BAP1 and ASXL1) are among the most frequently mutated epigenetic factors in human cancers. How PR-DUB establishes specificity for H2AK119Ub over other nucleosomal ubiquitination sites and how disease-associated mutations of the enzyme affect activity are unclear. Here, we determine a cryo-EM structure of human BAP1 and the ASXL1 DEUBAD in complex with a H2AK119Ub nucleosome. Our structural, biochemical, and cellular data reveal the molecular interactions of BAP1 and ASXL1 with histones and DNA that are critical for restructuring the nucleosome and thus establishing specificity for H2AK119Ub. These results further provide a molecular explanation for how >50 mutations in BAP1 and ASXL1 found in cancer can dysregulate H2AK119Ub deubiquitination, providing insight into understanding cancer etiology.

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We present the first complete genome sequence of the species Staphylococcus casei . Strain DSM 15096 was sequenced with a hybrid approach using Oxford Nanopore Technologies long-read sequencing and Illumina short-read sequencing. The assembled sequences produced a 2 808 898 bp chromosomal molecule containing 2705 predicted genes, plus eight plasmids.

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Introduction. Helicobacter pylori is highly polymorphic, and some strains are much more likely to cause disease than others. Biofilm formation can help bacteria to survive antibiotic treatment, immune attack and other stresses, promoting persistent infection.Hypothesis/Gap Statement. We hypothesized that H. pylori isolates from patients with more severe H. pylori-associated disease would be better at forming biofilms than isolates from patients with less severe disease.Aim. We initially aimed to determine whether or not the biofilm-forming ability of H. pylori isolates was associated with disease in the UK-based patients from whom the bacteria were isolated.Methodology. Biofilm-forming ability of H. pylori isolates was determined using a crystal violet assay on glass coverslips. The complete genome sequence of strain 444A was generated by hybrid assembly of Nanopore MinION and Illumina MiSeq data.Results. Although we found no associations between biofilm-forming ability of H. pylori and disease severity in patients, we discovered that strain 444A had particularly high biofilm-forming ability. This strain had been isolated from a patient with gastric ulcer disease and moderate to severe scores for H. pylori-induced histopathology. Analysis of the genome of the high biofilm-forming H. pylori strain 444A revealed that it possesses numerous biofilm- and virulence-associated genes and a small cryptic plasmid encoding a type II toxin-antitoxin system.Conclusion. There is substantial variation in biofilm-forming ability in H. pylori, but this was not significantly associated with disease severity in our study. We identified and characterized an interesting strain with high biofilm-forming ability, including generation and analysis of the complete genome.

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