Bacterial Fluorescence Research Articles

Versatile Dual Reporter to Identify Ribosome Pausing Motifs Alleviated by Translation Elongation Factor P.

Protein synthesis is influenced by the chemical and structural properties of the amino acids incorporated into the polypeptide chain. Motifs containing consecutive prolines can slow the translation speed and cause ribosome stalling. Translation elongation factor P (EF-P) facilitates peptide bond formation in these motifs, thereby alleviating stalled ribosomes and restoring the regular translational speed. Ribosome pausing at various polyproline motifs has been intensively studied using a range of sophisticated techniques, including ribosome profiling, proteomics, and in vivo screening, with reporters incorporated into the chromosome. However, the full spectrum of motifs that cause translational pausing in Escherichia coli has not yet been identified. Here, we describe a plasmid-based dual reporter for rapid assessment of pausing motifs. This reporter contains two coupled genes encoding mScarlet-I and chloramphenicol acetyltransferase to screen motif libraries based on both bacterial fluorescence and survival. In combination with a diprolyl motif library, we used this reporter to reveal motifs of different pausing strengths in an E. coli strain lacking efp. Subsequently, we used the reporter for a high-throughput screen of four motif libraries, with and without prolines at different positions, sorted by fluorescence-associated cell sorting (FACS) and identify new motifs that influence the translational efficiency of the fluorophore. Our study provides an in vivo platform for rapid screening of amino acid motifs that affect translational efficiencies.

Audit of Antimicrobial Prescribing Trends in 1447 Outpatient Wound Assessments: Baseline Rates and Impact of Bacterial Fluorescence Imaging.

Background/Objectives: In the field of wound care, the prescription of antibiotics and antimicrobials is haphazard and irrational, which has led to unchecked overprescribing. Recent Joint Commission guidelines mandate that hospital outpatient clinics develop and implement antimicrobial stewardship programs (ASPs). Yet few ASPs exist in wound clinics across the United States (US). Understanding baseline prescribing practices and rates in the US is a critical first step toward rational antimicrobial use and effective ASPs. Methods: This prospective study was conducted across eight outpatient wound clinics from January-December 2022. Data from consecutive patients attending single-time-point initial visits were recorded, including clinical findings, antimicrobial prescribing trends, and sampling practices. Results: A total of 1438 wounds were included; 964 were assessed by clinical examination (standard of care, SoC), and 474 by clinical examination plus fluorescence imaging. SoC patients were prescribed more concurrent medications on average than fluorescence patients (1.4 vs. 1 per patient). Prescriptions were preferentially topical in the fluorescence group (92% vs. 64%, p > 0.0001), and systemic antibiotics represented 36% of the single items prescribed under SoC (vs. 8% in fluorescence group p < 0.0001). Conclusions: Fluorescence imaging provided objective and actionable information at the bedside, which led to a decrease in the use of antibiotics. Real-time diagnostic technologies are essential in establishing a meaningful ASP.

Nitrogen-Doped Carbon Dots: A New Powerful Fluorescent Dye with Substantial Effect on Bacterial Cell Labeling.

Carbon dots (CDs)-minute carbon nanoparticles with remarkable luminescent properties, photostability, and low toxicity-show potential for various applications. CDs synthesized using citric acid and urea are the least toxic to biological environments. Here, we aimed to explore the effect of CDs synthesized using citric acid and urea at 50, 33, and 25% (CDs 1/1, 1/2, and 1/3, respectively) weight ratios in a microwave on bacterial cell fluorescence sensing and labeling. The nanoscale properties of CDs were investigated via transmission electron microscopy and dynamic light scattering particle size analysis. X-ray powder diffraction confirmed the graphitic structures of CDs. X-ray photoelectron spectroscopy revealed that the nitrogen content increased gradually with increasing urea ratios, indicating functional group changes. Transient photoluminescence decay periods demonstrated superior fluorescence intensity of CDs 1/3 under blue, green, and red lights. The use of CDs was notably more efficient than traditional methods in staining bacterial cells. Fluorescence microscopy of 10 g-positive and 10 g-negative bacteria revealed enhanced staining of Gram-positive strains, with CDs 1/3 presenting the best results. The CDs exhibited excellent photostability, maintaining poststaining fluorescence for 100 min, surpassing the performance of conventional dyes. CDs could serve as potential fluorescent dyes for the rapid discrimination of Gram-positive and Gram-negative bacteria.

Bacterial Fluorescence Imaging to Address Racial Inequities in Wound Infection Assessment.

Bacterial Fluorescence Imaging to Address Racial Inequities in Wound Infection Assessment.

Improving Wound Healing and Infection Control in Long-term Care with Bacterial Fluorescence Imaging.

High bacterial burden stalls wound healing and can quickly progress to infection and sepsis in complex, older-adult patients in long-term care (LTC) or skilled nursing facilities (SNFs). To investigate the outcomes of point-of-care fluorescence (FL) imaging (MolecuLight i:X) of bacterial loads, which are frequently asymptomatic, to inform customized wound treatment plans for patients in LTC/SNFs. In this retrospective pre/postinterventional cohort study, the authors compared the healing and infection-associated outcomes of 167 pressure injuries from 100 Medicare beneficiaries before and after implementation of FL imaging. Most patient demographics and wound characteristics did not differ significantly between the standard-of-care (SOC; n = 71 wounds) and FL (n = 96 wounds) cohorts. Significantly more wounds (+71.0%) healed by 12 weeks in the FL cohort (38.5%) versus the SoC cohort (22.5%). Wounds in the FL cohort also healed 27.7% faster (-4.8 weeks), on average, and were 1.4 times more likely to heal per Kaplan-Meier survival analysis (hazard ratio = 1.40; 95% CI, 0.90-2.12). Infection-related complications decreased by 75.3% in the FL cohort, and a significant shift from largely systemic to topical antibiotic prescribing was evidenced. Fluorescence-imaging-guided management of wounds significantly improved healing and infection outcomes in highly complex and multimorbid patients in LTC/SNFs. Proactive bacterial infection management via local treatments was enabled by earlier, objective detection. These reported outcome improvements are comparable to randomized controlled trials and cohort studies from less compromised, selectively controlled outpatient populations. Fluorescence imaging supports proactive monitoring and management of planktonic and biofilm-encased bacteria, improving patient care in a complex, real-world setting.

Determination of the Course of Cyan Fluorescence of Pseudomonas aeruginosa with a Handheld Bacterial Imaging Device.

Chronic wound infections are of clinical concern as they often lead to high rates of mortality and morbidity. A point-of-care handheld bacterial fluorescence imaging has been designed to detect the auto-fluorescent characteristics of most clinically relevant species of bacteria. This device causes most species of bacteria to exhibit red fluorescence due to the production of exoproduct porphyrins. One of the most significant contributors to the pathogenicity of chronic wounds is the pathogen Pseudomonas aeruginosa, and interestingly, this organism exhibits an additional unique cyan fluorescence signature. There is an over 90% positive predictive value that, when a chronic wound exhibits cyan fluorescence with the bacterial fluorescence imaging device, the wound will harbor P. aeruginosa. This project seeks to understand what genetic factor(s) contribute to the cyan phenotype observed.

Bis-butoxyphenyl dipyridine cation with high affinity for PF6−: Enhancement of solid fluorescence and inhibits Escherichia coli and Staphylococcus aureus activity through anion-π+ interactions

The contamination of anions and the overuse of antibiotics pose significant hazards to human health. Excess anions can cause serious damage to organisms and the natural environment, while the misuse of antibiotics has led to the emergence of multi-drug resistant bacteria. Therefore, it is important to develop multifunctional fluorescent sensing devices that integrate the rapid and sensitive detection of specific anions and that exhibit excellent antimicrobial activity against multi-resistant bacteria. This study describes the synthesis of three water-soluble cationic fluorescent molecules, DPPT-R (R = 5C, 6C, 7C). Their structures were characterized using 1H NMR spectroscopy, HRMS, and single crystal X-ray diffraction. Spectroscopic properties of the probes were recorded using UV–visible absorption and fluorescence spectroscopy. The recognition mode and mechanism of the fluorescent probes towards PF6− were investigated. Under aqueous solution conditions involves the PF6− induced aggregation of the probe via strong electrostatic interactions, leading to fluorescence quenching. Adjusting the alkyl chain length of the probe can enhance the sensitivity for the detection of the target anions. The detection limits are 0.892 μM, 0.786 μM and 0.444 μM for R = 5C, 6C, 7C, respectively. In addition, DPPT-R and DPPT-R@PF6 exhibit good inhibitory activity and fluorescence imaging capability against methicillin-resistant Staphylococcus aureus and multidrug-resistant Escherichia coli. Among them, the side chain of DPPT-R@PF6 with an almost 90° bend may be more advantageous for membrane insertion, thus enhancing its antimicrobial activity. In particular, after DPPT-R and PF6− form the DPPT-R@PF6 complex, the complex enhances its solid-state fluorescence, intensity of bacterial fluorescence imaging, and antibacterial activity through anion-π+ interactions. Therefore, this study can provide a new strategy and theoretical basis for the design and application of multifunctional fluorescent sensing materials and antibacterial molecules based on electrostatic interactions and anion-π+ interactions.

Water-soluble vitamin-E for enhancing fluorescence diagnosis in infected human dentine treated with NaOCl

IntroductionBacterial fluorescence methods are of interest in endodontics for informing endpoints for debridement. This study explored potential fluorescence quenching reversal effects of a water-soluble vitamin E conjugate (d-α-Tocopherol polyethylene glycol 1000 succinate, TPGS) when applied to polymicrobial biofilms grown on dentine that had been exposed to sodium hypochlorite (NaOCl) to cause quenching. MethodExtracted human teeth were debrided, embedded in transparent acrylic resin and sectioned. After smear layer removal, tooth dentine sections were inoculated with a polymicrobial inoculum, and cultured for 7 days to create biofilms. Samples (n = 8 per group) were exposed to 1 % or 4 % NaOCl for 2 or 4 min, and then treated with TPGS. Bacterial fluorescence readings under laser excitation at 655 nm were assessed over 10 min using a calibrated DIAGNOdent device. All data were assessed for normality (Kolmogorov-Smirnov test) and analysed with ANOVA followed by Bonferroni post-hoc tests. ResultsNaOCl at both concentrations quenched fluorescence readings of biofilms grown on dentine samples, with a maximal reduction of 40.4 % at 5 min after 4 % NaOCl. Treatment with TPGS gave faster recovery of fluorescence readings compared to the control at 5 and 10 min. ConclusionThe water-soluble antioxidant TPGS partially reversed fluorescence quenching caused by NaOCl. This agent may have value clinically for reducing the time needed for fluorescence readings to recover when NaOCl is used as an irrigant. This will facilitate more accurate assessment of endpoints for canal debridement.

Synthesis and performance study of novel antibacterial and anticorrosive polyurethane IPN

Polyaniline has been extensively used for the corrosion protection of polymeric materials. However, there have been few studies on its combined antimicrobial activity when used in combination with silver molybdate. In this study, we synthesized silver molybdate (Ag2MoO4) crystals using a homogeneous precipitation method. Polyaniline-dodecylbenzenesulfonic acid/castor oil(PANI-DBSA/CO) mixtures were prepared in an encapsulated state by in situ polymerisation. Subsequently, PANI/Ag2MoO4-PU interpenetrating polymer network(IPN) composite coatings(P/A-PU) were produced by dispersing Ag2MoO4 and PANI-DBSA/CO in polyurethane using ultrasonic dispersion. The composite coatings were characterized using Fourier transform infrared spectroscopy(FTIR), X-ray diffraction analysis(XRD), and transmission electron microscopy(TEM). The study confirmed that composite coatings with uniformly dispersed nanoparticles of PANI and Ag2MoO4 encapsulated by CO and epoxy resin E-44 grease exhibit improved physico-mechanical characteristics. The research also evaluated the effect of nanoparticles on the corrosion behaviour of the coatings using electrochemical impedance spectroscopy and kinetic potential polarisation experiments. The study investigated the antimicrobial properties of P/A-PU composite coatings on Escherichia coli(E.coli) and Staphylococcus aureus(S.aureus) by conducting bacterial growth curves, protein leakage, and fluorescence staining experiments. The aim was to establish a causal connection between the coated surfaces and the observed antimicrobial properties. The study demonstrated that incorporating IPN of polyaniline polyurethane into the coating composites improved their anticorrosive properties. Furthermore, the addition of silver molybdate at a mass ratio of 0.3 % resulted in a remarkable 99 % antimicrobial activity against E.coli and S.aureus.

A Diagnostic-Driven Prospective Clinical Study Evaluating the Combination of an Antibiofilm Agent and Negative Pressure Wound Therapy.

Each year, millions of Americans develop truncal pressure ulcers (PUs) which can persist for months, years, or until the end of life. Despite the negative impact on quality of life and escalating costs associated with PUs, there is sparse evidence supporting validated and efficacious treatment options. As a result, treatment is based on opinion and extrapolation from other wound etiologies. The ideal reconstructive plan maximizes the patient's nutritional status, incorporates the basic tenets of wound bed preparation (debridement, offloading, proper moisture balance, reduction of bacterial burden), and employs diagnostics to guide therapeutic intervention. The use of combination therapies can potentially overcome several of the barriers to wound healing. Negative pressure wound therapy (NPWT), a commonly used modality in the management of PUs, facilitates healing by stimulating the formation of granulation tissue and promoting wound contraction; however, NPWT alone is not always effective. Clinical studies examining microbial bioburden in PUs determined that most ulcers contain bacteria at levels that impede wound healing (>104 CFU/g). Thus, we hypothesized that adding an anti-microbial agent to decrease both planktonic and biofilm bacteria in the wound would increase the efficacy of NPWT. In this prospective study, twenty patients with recalcitrant PUs that previously failed NPWT were treated with a biofilm-disrupting agent (Blast-X, Next Science, Jacksonville, FL, USA) in combination with NPWT. Fluorescence imaging was used to follow bacterial burden and guide therapy. In total, 45% of the PUs reduced in size over the course of the four-week study, with a resolution of bacterial fluorescence in the NPWT dressing and wound bed seen in an average of three weeks. The combination of an antibiofilm agent and NPWT reduced bacterial levels and improved wound healing in recalcitrant PUs.

Bacterial fluorescence imaging as a predictor of skin graft integration in burn wounds

BackgroundSplit-thickness skin graft (STSG)11STSG: Split-thickness skin grafts integration rates are susceptible to improvement. Infection and/or biofilm should be appropriately addressed prior to grafting to improve the likelihood of graft-take. Incorporating technological aids such as fluorescence (FL)22FL: Fluorescence imaging (MolecuLight®), which accurately locates areas of bacterial loads above 104 CFU/gr, for graft site assessment and preparation could yield better outcomes. MethodsThis single-center, prospective observational study included adult burn patients with previously infected wounds that had been deemed clinically and microbiologically clean and were therefore candidates for grafting. Prior to grafting, a FL imaging assessment (blinded to the surgical team) localized areas positive for moderate-high bacterial loads (>104 CFU/gr). Intra-operatively, a standard swab sample from the recipient site was collected by the surgical team. Postoperatively, areas positive/negative for FL and areas of graft take and failure were overlapped and measured (cm2) over a 2D schematic. The performance and accuracy of FL imaging and swab sampling in relation to graft outcomes were assessed. Results38 patients were enrolled in the study. The mean total body surface area (TBSA)33TBSA: Total body surface area involvement was 14.5 ± 12.4 % [range 0.8 – 40.2 %]. 25/38 of the subjects enrolled had complete graft take while 13 had partial graft losses. There were no total losses. FL-imaging was positive in 100 % of losses versus 31 % (4/13) of the swab microbiology. FL-imaging was found to have a sensitivity of 86 %, specificity of 98 %, PPV of 72 %, NPV of 99 %, and an accuracy of 94 % for predicting any type or range of graft loss in the entire cohort. Meanwhile, the sensitivity of microbiology from swab samples was 30 %, with a specificity of 76 %. ConclusionsFL imaging is an accurate method for assessing recipient sites and predicting the outcome of a skin graft among burn patients. These findings suggest that FL imaging can inform better decision-making surrounding grafts that may lead to better outcomes. Level of evidenceLevel IIA, Therapeutic study

Space-confined nanozyme with cascade reaction based on PCN-224 for synergistic bacterial infection treatment and NIR fluorescence imaging of wound bacterial infections

The rapid development of bacterial infections has become a significant global health challenge. The overuse or prolonged use of antibiotics is considered a major contributor to the development of bacterial resistance. Therefore, there is an urgent need to develop a new therapeutic approach to combat infectious diseases caused by bacteria. In this study, we demonstrate the effectiveness of a novel strategy for synergistic antibacterial therapy and fluorescence imaging of wound bacterial infections through the nano-confined construction of LL-37-HA-Au-Pt@PCN-224 nanozymes. The nanozyme is composed of Au NPs and Pt NPs confined to PCN-224 through in situ reduction, with its outer layer modified with HA and LL-37. The nanozyme catalyzes the conversion of low-concentration H2O2 into O2 and •OH, with the generated O2 further catalyzed into 1O2 by PCN-224. In vivo imaging experiments demonstrate that LL-37-HA-Au-Pt@PCN-224 labeled with near-infrared fluorescent dye MPA can effectively realize fluorescence imaging of wound bacterial infections and real-time monitoring of the wound healing process. In vitro and in vivo antibacterial experiments reveal that LL-37-HA-Au-Pt@PCN-224 enhances the healing efficiency of bacteria-infected wounds due to its synergistic antibacterial effect. The multifunctional nanozyme offers various synergistic antibacterial therapies, including antimicrobial peptides, peroxide nanozymes, hydrogen peroxide nanozymes, and photodynamic therapy. Moreover, it provides real-time wound imaging and monitoring of the wound healing process, while maintaining high biocompatibility. Overall, this study presents a promising perspective on the potential use of nanozymes as antibacterial agents against wound bacterial infections in the future.

Development and utilization of new O2-independent bioreporters.

Fluorescent proteins have revolutionized science since their discovery in 1962. They have enabled imaging experiments to decipher the function of proteins, cells, and organisms, as well as gene regulation. Green fluorescent protein and all its derivatives are now standard tools in cell biology, immunology, molecular biology, and microbiology laboratories around the world. A common feature of these proteins is their dioxygen (O2)-dependent maturation allowing fluorescence, which precludes their use in anoxic contexts. In this work, we report the development and in cellulo characterization of genetic circuits encoding the O2-independent KOFP-7 protein, a flavin-binding fluorescent protein. We have optimized the genetic circuit for high bacterial fluorescence at population and single-cell level, implemented this circuit in various plasmids differing in host range, and quantified their fluorescence under both aerobic and anaerobic conditions. Finally, we showed that KOFP-7-based constructions can be used to produce fluorescing cells of Vibrio diazotrophicus, a facultative anaerobe, demonstrating the usefulness of the genetic circuits for various anaerobic bacteria. These genetic circuits can thus be modified at will, both to solve basic and applied research questions, opening a highway to shed light on the obscure anaerobic world.IMPORTANCEFluorescent proteins are used for decades, and have allowed major discoveries in biology in a wide variety of fields, and are used in environmental as well as clinical contexts. Green fluorescent protein (GFP) and all its derivatives share a common feature: they rely on the presence of dioxygen (O2) for protein maturation and fluorescence. This dependency precludes their use in anoxic environments. Here, we constructed a series of genetic circuits allowing production of KOFP-7, an O2-independant flavin-binding fluorescent protein. We demonstrated that Escherichia coli cells producing KOFP-7 are fluorescent, both at the population and single-cell levels. Importantly, we showed that, unlike cells producing GFP, cells producing KOFP-7 are fluorescent in anoxia. Finally, we demonstrated that Vibrio diazotrophicus NS1, a facultative anaerobe, is fluorescent in the absence of O2 when KOFP-7 is produced. Altogether, the development of new genetic circuits allowing O2-independent fluorescence will open new perspective to study anaerobic processes.

An objective comparative study of non-surgical cleansing techniques and cleanser types in bacterial burden management.

Cleansing is a vital component of effective wound hygiene and biofilm management, often accomplished through vigorous mechanical action or through soaking with moistened gauze. In the present study, a quantitative comparison of the effectiveness of different cleansing techniques and solutions in removing bacteria was conducted on 71 chronic wounds using bacterial fluorescence imaging as a real-time diagnostic for moderate to high bacterial loads. Vigorous gauze cleansing for 30 s proved most effective by reducing bacterial fluorescence by 33.99%, surpassing 10-min soaking in bacterial reduction (13.24%). Among different cleansers, no statistically significant differences in effectiveness were observed, but povidone-iodine showed the strongest trend towards bacterial reduction. Sub-analysis highlighted the superiority of antiseptic cleansers over saline and gentle soap (-33.30% vs. -1.80% bacterial reduction respectively). Five percent acetic acid was also shown to be more effective in removing specific bacterial strains (Pseudomonas aeruginosa). Findings from studies like this contribute to refining wound hygiene guidelines and clinical algorithms for bacterial and biofilm management.

Time-series analysis of bacterial counts and fluorescence properties in hot-filled tofu during storage

Time-series analysis of bacterial counts and fluorescence properties in hot-filled tofu during storage

Integrating Point-of-Care Bacterial Fluorescence Imaging-Guided Care with Continued Wound Measurement for Enhanced Wound Area Reduction Monitoring.

This prospective observational study investigated wound area reduction (WAR) outcomes in a complex wound population composed of non-healing acute and chronic wounds. The relationship between bacterial autofluorescence signals and WAR was investigated. Area measurements were collected both manually and digitally, and both methods were compared for accuracy. Twenty-six participants with 27 wounds of varying etiologies were observed twice weekly for two weeks. Digital wound measurement, wound bacterial status assessment, and targeted debridement were performed through a point-of-care fluorescence imaging device (MolecuLight® i: X, MolecuLight Inc, Toronto, Canada). The wound area reduction (WAR) rate was calculated using baseline and last visit measurements. Statistical analyses, including t-tests, Fisher exact tests, the Wilcoxon signed rank test for method comparison, and ANOVA for bacterial subgroups, were applied as pertinent. The overall average WAR was -3.80 cm2, or a decrease of 46.88% (manual measurement), and -2.62 cm2, or a 46.05% decrease (digital measurement via MolecuLight® device). There were no statistically significant differences between the WAR of acute and chronic wounds (p = 0.7877). A stepwise correlation between the WAR and bacterial status classification per fluorescence findings was observed, where persistent bacteria resulted in worse WAR outcomes. An overestimation of wound area by manual measurement was 23% on average. Fluorescence imaging signals were linked to WAR outcome and could be considered predictive. Wounds exhibiting bacterial loads that persisted at the end of the study period had worse WAR outcomes, while those for which management was able to effectively remove them demonstrated greater WAR. Manual measurement of the wound area consistently overestimated wound size when compared to digital measurement. However, if performed by the same operator, the overestimation was uniform enough that the WAR was calculated to be close to accurate. Notwithstanding, single wound measurements are likely to result in overestimation.

The contribution of IL-17A-dependent low LCN2 levels to Helicobacter pylori infection: Insights from clinical and experimental studies

BackgroundHelicobacter pylori (H. pylori) infection is a common bacterial infection that is widespread globally. It is crucial to comprehend the molecular mechanisms that underlie the infection caused by H. pylori in order to devise successful therapeutic approaches. The objective of this study was to examine the involvement of Lipocalin-2 (LCN2) in the development of H. pylori infection. MethodsLCN2 expression levels in human gastric mucosa and H. pylori-infected mouse models were analyzed using quantitative PCR and immunohistochemistry methods. The effects of LCN2 on the attachment of H. pylori to gastric mucosa cells were assessed using bacterial culture and fluorescence intensity tests. To investigate the correlation between LCN2, CCL20, and IL-17A, we performed gene expression analysis and measured serum levels. ResultsThe findings indicated an increase in LCN2 levels in the gastric mucosa of both patients and mice infected with H. pylori. Blocking the natural LCN2 resulted in an increased attachment of H. pylori to cells in the gastric mucosa. In addition, we noticed that reduced levels of LCN2 promoted the attachment of H. pylori to cells in the gastric mucosa. Furthermore, H. pylori-infected patients exhibited increased expression of both LCN2 and CCL20, and there was a positive correlation between serum levels of CCL20 and LCN2. LCN2 expression was found to depend on the presence of IL-17A, and inhibiting IL-17A led to a higher H. pylori colonization. ConclusionThe persistence of H. pylori infection is facilitated by the presence of low levels of LCN2, which is dependent on IL-17A. This finding offers valuable perspectives for the development of novel therapeutic approaches for H. pylori infection.

Is my wound infected? A study on the use of hyperspectral imaging to assess wound infection.

Clinical signs and symptoms (CSS) of infection are a standard part of wound care, yet they can have low specificity and sensitivity, which can further vary due to clinician knowledge, experience, and education. Wound photography is becoming more widely adopted to support wound care. Thermography has been studied in the medical literature to assess signs of perfusion and inflammation for decades. Bacterial fluorescence has recently emerged as a valuable tool to detect a high bacterial load within wounds. Combining these modalities offers a potential objective screening tool for wound infection. A multi-center prospective study of 66 outpatient wound care patients used hyperspectral imaging to collect visible light, thermography, and bacterial fluorescence images. Wounds were assessed and screened using the International Wound Infection Institute (IWII) checklist for CSS of infection. Principal component analysis was performed on the images to identify wounds presenting as infected, inflamed, or non-infected. The model could accurately predict all three wound classes (infected, inflamed, and non-infected) with an accuracy of 74%. They performed best on infected wounds (100% sensitivity and 91% specificity) compared to non-inflamed (sensitivity 94%, specificity 70%) and inflamed wounds (85% sensitivity, 77% specificity). Combining multiple imaging modalities enables the application of models to improve wound assessment. Infection detection by CSS is vulnerable to subjective interpretation and variability based on clinicians' education and skills. Enabling clinicians to use point-of-care hyperspectral imaging may allow earlier infection detection and intervention, possibly preventing delays in wound healing and minimizing adverse events.

Identifying infection in chronic wounds in a community setting: A systematic review of diagnostic test accuracy studies.

To determine the diagnostic accuracy of different methods currently available to identify infection in chronic wounds applicable to adult patients in a community setting. Systematic review of diagnostic test accuracy studies. Two authors independently completed screening, data extraction and quality and bias assessments (QUADAS2). Eligible studies compared a method (index test) for detecting infection (diagnosis of interest) with microscopy and culture of either deep tissue biopsy or wound swab (reference test) in adult patients with wounds of >4 weeks duration (participants). The results were synthesized narratively. We systematically searched CINAHL, Embase and Medline from 2011 to April 2022. Four studies were included, all recruiting from secondary care wound clinics. Two studies assessed the diagnostic accuracy of Moleculight i:X, a bacterial fluorescence imaging device against deep tissue biopsy culture. One study assessed the diagnostic accuracy of the elevation of various enzymes detected in wound fluid against wound swab microscopy of culture. One study assessed the diagnostic accuracy of bacterial protease activity against wound swab microscopy and culture. Sensitivities of these methods ranged from 50 to 75% and specificities from 47 to 100%. Only a small number of studies were included in this systematic review due to our strict inclusion criteria. We have not identified any methods for diagnosing infection in chronic wounds with either a sufficient quality of evidence to recommend their use in community settings at present. Further research is needed to develop and evaluate appropriate diagnostics for this purpose. This study highlights the paucity of research into wound diagnostics in a community setting and should prompt further research in this area. Accurate diagnostic tests have the potential to improve community-based wound care by optimizing antibiotic use and potentially improving healing time. PRISMA-DTA checklist. The PPI group for the NIHR Community Healthcare MIC were supportive of this topic of work.

A case of canine intestinal malakoplakia

Malakoplakia is a rare chronic granulomatous disease usually affecting the urinary bladder and other locations. In humans, the gastrointestinal tract is the second most common location but there are no reports of intestinal malakoplakia in animals. A 10-month-old female French Bulldog was presented with chronic haemorrhagic diarrhoea and anorexia with normochromic-normocytic anaemia and hypoalbuminaemia. Grossly, there was mucosal thickening and ulceration of the caecum, colon and rectum. Microscopically, transmural sheets of foamy macrophages were seen in these tissues. Macrophages were periodic acid–Schiff, vimentin and ionized calcium-binding adaptor molecule 1 positive and contained von Kossa- and Prussian blue-positive Michaelis–Gutmann bodies. Giemsa staining revealed rod-shaped bacterial colonies and fluorescence in-situ hybridization demonstrated Escherichia coli within macrophages. This is the first reported case of intestinal malakoplakia in domestic animals. Pathological features of intestinal malakoplakia share many similarities with ulcerative histiocytic colitis in dogs but it is unclear if they are different forms of the same pathological process or distinct entities.