Urine biomarkers individually and as a consensus model show high sensitivity and specificity for detecting UTIs.

BACKGROUND: Current diagnoses of urinary tract infection (UTI) by standard urine culture (SUC) has significant limitations in sensitivity, especially for fastidious organisms, and the ability to identify organisms in polymicrobial infections. The significant rate of both SUC "negative" or "mixed flora/contamination" results in UTI cases and the high prevalence of asymptomatic bacteriuria indicate the need for an accurate diagnostic test to help identify true UTI cases. This study aimed to determine if infection-associated urinary biomarkers can differentiate definitive UTI cases from non-UTI controls. METHODS: Midstream clean-catch voided urine samples were collected from asymptomatic volunteers and symptomatic subjects ≥ 60 years old diagnosed with a UTI in a urology specialty setting. Microbial identification and density were assessed using a multiplex PCR/pooled antibiotic susceptibility test (M-PCR/P-AST) and SUC. Three biomarkers [neutrophil gelatinase-associated lipocalin (NGAL), and Interleukins 8 and 1ß (IL-8, and IL-1ß)] were also measured via enzyme-linked immunosorbent assay (ELISA). Definitive UTI cases were defined as symptomatic subjects with a UTI diagnosis and positive microorganism detection by SUC and M-PCR, while definitive non-UTI cases were defined as asymptomatic volunteers. RESULTS: We observed a strong positive correlation (R2 > 0.90; p < 0.0001) between microbial density and the biomarkers NGAL, IL-8, and IL-1ß for symptomatic subjects. Biomarker consensus criteria of two or more positive biomarkers had sensitivity 84.0%, specificity 91.2%, positive predictive value 93.7%, negative predictive value 78.8%, accuracy 86.9%, positive likelihood ratio of 9.58, and negative likelihood ratio of 0.17 in differentiating definitive UTI from non-UTI cases, regardless of non-zero microbial density. NGAL, IL-8, and IL-1ß showed a significant elevation in symptomatic cases with positive microbe identification compared to asymptomatic cases with or without microbe identification. Biomarker consensus exhibited high accuracy in distinguishing UTI from non-UTI cases. CONCLUSION: We demonstrated that positive infection-associated urinary biomarkers NGAL, IL-8, and IL-1ß, in symptomatic subjects with positive SUC and/or M-PCR results was associated with definitive UTI cases. A consensus criterion with ≥ 2 of the biomarkers meeting the positivity thresholds showed a good balance of sensitivity (84.0%), specificity (91.2%), and accuracy (86.9%). Therefore, this biomarker consensus is an excellent supportive diagnostic tool for resolving the presence of active UTI, particularly if SUC and M-PCR results disagree.

Elevated UTI Biomarkers in Symptomatic Patients with Urine Microbial Densities of 10,000 CFU/mL Indicate a Lower Threshold for Diagnosing UTIs.

The literature lacks consensus on the minimum microbial density required for diagnosing urinary tract infections (UTIs). This study categorized the microbial densities of urine specimens from symptomatic UTI patients aged ≥ 60 years and correlated them with detected levels of the immune response biomarkers neutrophil gelatinase-associated lipocalin (NGAL), interleukin-8 (IL-8), and interleukin-1-beta (IL-1ß). The objective was to identify the microbial densities associated with significant elevation of these biomarkers in order to determine an optimal threshold for diagnosing symptomatic UTIs. Biobanked midstream voided urine samples were analyzed for microbial identification and quantification using standard urine culture (SUC) and multiplex-polymerase chain reaction (M-PCR) testing, while NGAL, IL-8, and IL-1ß levels were measured via enzyme-linked immunosorbent assay (ELISA). NGAL, IL-8, and IL-1ß levels were all significantly elevated at microbial densities ≥ 10,000 cells/mL when measured via M-PCR (p < 0.0069) or equivalent colony-forming units (CFUs)/mL via SUC (p < 0.0104) compared to samples with no detectable microbes. With both PCR and SUC, a consensus of two or more elevated biomarkers correlated well with microbial densities > 10,000 cells/mL or CFU/mL, respectively. The association between ≥10,000 cells and CFU per mL with elevated biomarkers in symptomatic patients suggests that this lower threshold may be more suitable than 100,000 CFU/mL for diagnosing UTIs.

A test combining multiplex-PCR with pooled antibiotic susceptibility testing has high correlation with expanded urine culture for detection of live bacteria in urine samples of suspected UTI patients.

We evaluated whether multiplex polymerase chain reaction (M-PCR) detects viable micro-organisms by comparing micro-organism identification with standard urine culture (SUC) and expanded quantitative urine culture (EQUC). Of the 395 organisms detected by M-PCR, EQUC detected 89.1% (p = 0.10), whereas SUC detected 27.3% (p < 0.0001 vs. M-PCR and p < 0.0001 vs EQUC). M-PCR identified 260 nonfastidious bacteria, EQUC detected 96.5% (p = 0.68), whereas SUC detected 41.5% (p < 0.0001). Common nonfastidious bacteria missed by SUC included Escherichia coli (72.5% detected), Klebsiella pneumoniae (66.7% detected), Enterococcus faecalis (34.6% detected) and Enterococcus faecium (0% detected). M-PCR identified 135 fastidious bacteria and EQUC 101 (74.8%, p = 0.01), whereas SUC failed to detect any (0%, p < 0.0001). Clinical samples evaluated using EQUC and M-PCR yielded very similar findings, indicating that most microbes identified by M-PCR represented viable organisms, and validating M-PCR as a diagnostic tool for UTIs.

Quantitative multiplex polymerase chain reaction in copies ml-1 linearly correlates with standard urine culture in colonies ml-1 for urinary tract infection (UTI) pathogens.

Standard urine culture (SUC) is the current standard method for confirmation of a urinary tract infection (UTI). SUC identifies microorganisms in urine samples and semi-quantifies these as colony-forming units (CFUs) ml-1. In contrast, quantitative multiplex polymerase chain reaction (q-MPCR) is a culture-independent assay in which the microbes are quantified by targeting genomic sequences and reported as cells ml-1, calculated from copies ml-1. Using serial dilutions within the 104-105 cells ml-1 range, the usual reporting range of SUC, this study compared the quantification results based on SUC and q-MPCR for four uropathogens with the control hemocytometer counts. The results revealed a linear relationship and a 1:1 correlation between the q-MPCR and SUC results. Additional q-MPCR quantification of 36 uropathogenic non-fastidious and fastidious bacteria and yeast indicated a reproducible linear correlation in a 1:1 manner with the control counts over a range of cell densities (103-106 cells ml-1). The results confirm that the quantifications by q-MPCR in cells ml-1 and by SUC in CFUs ml-1 are comparable and answer to the lingering question of how the results of these two methods correlate. Moreover, q-MPCR provided accurate quantification of various microorganisms over wider cell density ranges without the time required for microbial growth.

Bigger babies: what happens in real practice in a non-academic UK center? Detection accuracy and outcomes with induction.

BACKGROUND: There is emerging evidence of improved outcomes with induction of labour for pregnancies in which the baby is thought to be large. This trial identifies scan accuracy and the effect of intervention for pregnancies complicated by suspected large for gestational age (LGA) on customized chart outside an academic center. METHODS: This is a retrospective cohort study of 3 groups of induced pregnancies; women with a suspected LGA fetus, women with diabetes (DM) and a control group (C) of women that underwent induction of labour on or after 280 days gestation. Data collection and analysis were prespecified. Scan accuracy and outcomes between the cohorts were compared. RESULTS: Over 1 year there were 845 cases: LGA (128), DM (116) and control cases (601). Mean birthweights differed significantly. PPV of EFW for birthweight >90th centile on GROW chart, WHO chart, and >4 kg was 0.35-0.40. Projected birthweight of >4 kg significantly better predicted itself (AUROC 0.70, 0.74 and 0.80). Mean scan error was -5.2% and +15.6% for DM and LGA. Shoulder dystocia and neonatal morbidity were not increased in LGA despite the significant increase in AVD 28/128, 21.9% vs. 99/601, 16.5%, aOR 2.20 (1.07-4.5). SVD was significantly less likely LGA vs. C at 69/128, 53.9% vs. 413/601, 68.7% aOR 0.38 (95% CI: 0.21-0.70). CONCLUSIONS: Third trimester EFW for bigger babies was poorly predictive of macrosomia. Fetal outcomes were good but women selected and induced as LGA had higher rates of hemorrhage and intervention.

Are trainees working in obstetrics and gynecology confident and competent in the care of frail gynecological oncology patients?

INTRODUCTION: Older patients undergoing cancer surgery are at increased risk of post-operative complications, prolonged hospital stay, and mortality. Identification of frailty can help predict patients at high risk of peri-operative complications and allow a collaborative, multidisciplinary team approach to their care. A survey was conducted to assess the confidence and knowledge of trainees in obstetrics and gynecology regarding identification and management of peri-operative issues encountered in frail gynecological oncology patients. METHODS: A web-based survey was distributed via the Audit and Research in Gynaecological Oncology (ARGO) collaborative and UK Audit and Research Collaborative in Obstetrics and Gynaecology (UKARCOG) . The survey on the management of frail peri-operative patients was disseminated to doctors-in-training (trainees) working in obstetrics and gynecology in the United Kingdom (UK) and Ireland. Specialty (ST1-7), subspecialty, and general practice trainees, non-training grade doctors, and foundation year doctors currently working in obstetrics and gynecology were eligible. Consultants were excluded. Study data were collected using REDCAP software hosted at the University of Manchester. Responses were collected over a 6-week period between January and February 2020. RESULTS: Of the 666 trainees who participated, 67% (425/666) reported inadequate training in peri-operative management of frail patients. Validated frailty assessment tools were used by only 9% (59/638) of trainees and less than 1% (4/613) were able to correctly identify all the diagnostic features of frailty. Common misconceptions included the use of chronological age and gender in frailty assessments. The majority of trainees (76.5%, 448/586) correctly answered a series of questions relating to mental capacity; however, only 6% (36/606) were able to correctly identify all three diagnostic features of delirium. A total of 87% (495/571) of trainees supported closer collaboration with geriatricians and a multidisciplinary approach. CONCLUSIONS: Obstetrics and gynecology trainees reported inadequate training in the peri-operative care of frail gynecological oncology patients, and overwhelmingly favored input from geriatricians. Routine use of validated frailty assessment tools may aid diagnosis of frailty in the peri-operative setting. There is an unmet need for formal education in the management of frail surgical patients within the UK and Irish obstetrics and gynecology curriculum.

The Evolution of Animal Models in Wound Healing Research: 1993-2017.

Significance: Wound healing is a complex and dynamic series of events influenced by a variety of intrinsic and extrinsic factors. Problematic wounds, particularly chronic wounds and pathologic scars, remain clinically significant burdens. Modeling physiologic and aberrant wound repair processes using in vitro or in vivo models have contributed to Advances in Wound Care (AWC); however, the fidelity of each model used, particularly with respect to its species-specific limitations, must be taken into account for extrapolation to human patients. Twenty-five years of wound healing models published in Wound Repair and Regeneration (1993-2017) and AWC (2012-2017) were collected and analyzed to determine trends in species utilization and models used. Recent Advances: In 25 years, 1,521 original research articles utilizing one or more wound models were published (total of 1,665 models). Although 20 different species were used over the course of 25 years, 5 species were most commonly utilized: human, mouse, rat, pig, and rabbit. In vivo modeling was used most frequently, followed by in vitro, ex vivo, and in silico modeling of wound healing processes. Critical Issues: A comparison of articles from 1993 to 1997 and 2013 to 2017 periods showed notable differences in model and species usage. Experiments utilizing mouse and human models increased, while the usage of pig models remained constant, rabbit and rat models declined in the more recent time period examined compared to the time period two decades before. Future Directions: This analysis shows notable changes in types of models and species used over time which may be attributed to new knowledge, techniques, technology, and/or reagents. Explorations into mechanisms of limb regeneration and wound healing of noncutaneous tissues have also contributed to a shift in modeling over time. Changes within the journals (i.e., page expansion and increased rejection rates), research funding, and model expense may also influence the observed shifts.

The Origin and Evolution of the HARMONIC® Scalpel.

When Jacques and Pierre Curie first researched ultrasonic energy and piezoelectric effects in the 1880s, they likely had no idea of the profound impact it would eventually have on surgical patients. Today in operating rooms around the world, ultrasonic energy is used for tissue manipulation, dissection, cutting, and coagulation. Surgeons including but not limited to the specialties of gynecology, general surgery, colorectal, thoracic, breast, and bariatric have activated ultrasonic energy in thousands of patients. As a mainstay surgical energy device, patients have benefited from the ultrasonic versatility of its cutting and coagulating effects. The ability of ultrasonic energy to be used near vital organs with precision by adjusting for tissue tension, power settings, and activation time has accounted for its safety and clinical outcomes. This overview of the mechanics of ultrasonic energy and the evolution of the HARMONIC® (UltraCision, Providence, Rhode Island, now owned by Ethicon Endo-Surgery, Inc., Cincinnati, Ohio) surgical tools since 1988 provides readers an understanding of this energy platform and its distinct advantages. Clinical implications of key research and clinical studies are explored and discussed with a focus on patient and surgical outcomes. Research in a variety of fields and tissues is presented with a special emphasis on the gynecological patient.

Itching for Knowledge About Wound and Scar Pruritus.

Chronic itch continues to be a problem that plagues millions of humans and animals. Pruritus has a negative impact on patient quality of life and many patients experience sleep deprivation, anxiety, and depression, similar to patients with chronic pain. This review provides an overview of clinical pruritus research with special emphasis on itch that wound care providers may see. In addition, the need for using multifactorial questionnaires for better research in pruritus is summarized. Similarities and differences in itch characteristics, triggers, and relievers in various patient populations are discussed. A brief overview of itch receptors and pathways is provided to help the reader better understand the complexity of the resultant itch sensation. Also, some nonpharmacological and pharmacological antipruritic therapies and their mechanisms of action are included.

Equine exuberant granulation tissue and human keloids: a comparative histopathologic study.

OBJECTIVE: To compare histopathologic features of a fibroproliferative disorder in horses (exuberant granulation tissue-EGT) and people (keloid). SAMPLE POPULATION: Archival tissue samples of EGT (n = 8) and keloid (12). METHODS: After automated hematoxylin and eosin, histochemical (Gomori trichrome, Verhoeff-van Gieson elastin) and immunohistochemical (vimentin, α-smooth muscle actin, CD34, CD68, CD117) stainings, tissue sections were evaluated using a semi-quantitative grading scale for presence or absence of ulceration, keloidal collagen, myofibroblasts, and elastic fibers as well as degree of inflammation, fibrosis, vascularity, and orientation of collagen fibers. RESULTS: Superficial dermis and deep dermis of both horses and people had increased numbers of haphazardly oriented thickened collagen fibers; however, only keloids contained "keloidal" collagen. Fibroblast numbers were markedly increased in both groups but only EGT had myofibroblasts. Minimal vascularity was observed in the deep dermis of both groups. The superficial dermis in EGT was characterized by small vessels within immature granulation tissue. Macrophages and mast cells were infrequently found in both groups but polymorphonuclear cells were markedly increased in EGT. CONCLUSIONS: Humans and horses are the only mammals known to naturally develop excessive granulation during wound healing; however, similarities and differences between fibroblast populations and associated collagen have not been reported. Inflammatory response may contribute to observed differences in the cellular populations, with EGT possessing markedly increased myofibroblasts, small vessels, and acute inflammatory cells compared with keloids. Further work is warranted to develop common treatment strategies for these fibroproliferative conditions.

Preclinical Models of Wound Healing: Is Man the Model? Proceedings of the Wound Healing Society Symposium.

SIGNIFICANCE: A review of therapeutic effects in preclinical and clinical studies suggests that concordance between large animal (pig=78%), small laboratory animal (53%) and in vitro (57%) results with those observed in humans is only partial. Pig models of wound healing provide major advantages over other animal models. Since the vast majority of wound-healing research is done in rodents and in vitro, the low concordance rate is a significant impediment to research that will have any clinical impact. CRITICAL ISSUES: To generate clinically relevant experimental data, hypothesis generation should begin, or at least involve human wound tissue samples. Such tissue could be used to test a predetermined hypothesis generated based on, say, murine data. Alternatively, such tissue could be analyzed using high-throughput cell biology techniques (e.g., genomics, proteomics, or metabolomics) to identify novel mechanisms involved in human wounds. Once the hypothesis has been formulated and confirmed using human samples, identification of these same mechanisms in animals represents a valid approach that could be used for more in-depth investigations and experimental manipulations not feasible with humans. FUTURE DIRECTIONS: This consensus statement issued by the Wound Healing Society symposium strongly encourages all wound researchers to involve human wound tissue validation studies to make their animal and cell biology studies more translationally and clinically significant.

Assessment of pruritus characteristics and impact on burn survivors.

The goal of this cross-sectional study was to characterize and describe persistent postburn pruritus. Cause and treatment of postburn itch is elusive. It has been suggested that burn survivors with persistent pruritus should be divided into acute itch (≤6 months postinjury) and chronic itch (>6 months postinjury) because the cause of itch may be different. Cross-sectional data of itch characteristics reported here are from the baseline data of a prospective, randomized, double-blind, controlled pilot study of 23 subjects with frequent and bothersome postburn pruritus. Subjects self-completed validated scales for variables of itch sensation, affect of itch, and severity. Variables of quality of life, frequency, pain and itch intensity, skin condition, scar, and medication were also recorded. Itch frequency revealed that 87% of subjects experienced itching daily, 96% experienced three or more episodes a day, and 52% had episode durations lasting 5 to 30 minutes per incidence. Itch was reported as unbearable by 94% of subjects with chronic itch and by 86% of subjects with acute itch, whereas bothersome was 88 and 100%, respectively. Itch sensation dimension of stinging was 74% in both acute and chronic itch subjects. Crawling and burning sensations were often severe. Potential itch triggers and relief activities were identified. Differences in sensory and affective itch components were detected between acute and chronic itch subjects. Combinations of itch mechanisms probably contribute to the development of and changes in pruritus. Characterizing the sensation and affective itch dimensions in conjunction with inflammation, burn injury, recovery, scar maturation, medication, and psychological status should better elucidate postburn itch.

Double-blind, randomized, pilot study assessing the resolution of postburn pruritus.

The objective of this study was to evaluate whether Provase®, a nonprescription moisturizer with a blend of protease enzymes, would reduce postburn itching in adult burn survivors relative to a control moisturizer. This was a prospective, single-center, double-blinded, pilot study where 23 burn survivors were randomized to either the treatment group, who applied Provase, or the control group, who applied the base moisturizer used in Provase every 8 hours for 4 weeks. Twelve were randomized to the treatment and 11 to the control groups with 9 participants in each group completing the study. There was no difference between groups with respect to gender, ethnicity, causative factor, TBSA burned, or time postinjury. Participant's pruritus and scar were reevaluated on a weekly basis for 4 consecutive weeks. Relative to baseline, there was a significant reduction of itch duration in minutes at weeks 3 and 4, the number of days per week that itch was experienced at weeks 2, 3, and 4, and the number of itch episodes per day at week 2 for the treatment group. The itch TBSA reduced significantly relative to baseline for the treatment group at week 1, 2, and 3. The affective itch characteristics were significantly reduced for the treatment group for bothersome at weeks 1, 2, 3, and 4; for annoying at week 4; and for unbearable at weeks 2, 3, and 4. Although this was a pilot study and not powered for statistical differences, there were statistically significant differences for itch duration, weekly frequency, itch episodes per day, itch TBSA, and reported affective burden of itch after treatment. Further investigation is recommended with a larger sample size treated for a longer period of time where participants are stratified based on acute or chronic itch.

Use of platelet rich plasma gel on wound healing: a systematic review and meta-analysis.

OBJECTIVE: Autologous platelet rich plasma is an advanced wound therapy used in hard-to-heal acute and chronic wounds. To better understand the use and clinical outcomes of the therapy, a systematic review of the published literature in cutaneous wounds was performed. METHODS: Electronic and hand searches for randomized controlled trials and comparative group studies using platelet rich plasma therapy in cutaneous wounds and published over the last 10 years was conducted. Eligible studies compared the treatment to standard care or other interventions. All citations were screened and eligible studies were assessed for validity, quality, and bias using accepted scoring methods. The primary outcomes were effect of platelet rich plasma and control wound care on wound healing and related healing measurements. Secondary outcomes related to healing such as infection, pain, exudate, adverse events, and quality of life were also considered. The meta-analysis utilized appropriate statistical methods to determine the overall treatment effect on chronic and acute wound healing and infection. RESULTS: The search terms resulted in 8577 citations and after removing duplicates and screening for protocol eligibility, a total of 24 papers were used. The meta-analysis of chronic wound studies revealed platelet rich plasma therapy is significantly favored for complete healing. The meta-analysis of acute wounds with primary closure studies demonstrated that presence of infection was reduced in platelet rich plasma treated wounds. CONCLUSIONS: This systematic review and meta-analysis of platelet rich plasma therapy in cutaneous wounds showed complete and partial wound healing was improved compared to control wound care.

Protein degradation and protection observed in the presence of novel wound dressing components.

Chronic wounds typically have excessive levels of matrix metalloproteinases (MMPs) and proinflammatory cytokines that impair healing. Reducing these detrimental proteins may be key to healing chronic wounds. Proprietary protease blends were formulated specifically to degrade excessive amounts of proinflammatory factors that could prevent wound healing. Applications of protease-containing wound dressings to acute and chronic wounds have been observed clinically to resolve inflammation and appear to aid healing. The purpose of this study was to test in vitro a deliberate blend of proteases for the ability to deactivate or activate known proteins associated with inflammation or healing. Purified human target proteins were incubated with test and control solutions and samples removed at various time points. Blinded samples were tested using a novel infrared protein multiplex sandwich-ELISA-type array technique. Many proinflammatory proteins such as MMPs, cytokines and chemokines were susceptible to degradation. Many proteins such as growth factors, cytokines and TIMP1 were resistant to degradation. Not all proinflammatory proteins were deactivated. Family protein structure did not appear to affect susceptibility to degradation or deactivation. These results suggest that specific protease containing wound dressings appear to reduce multiple detrimental components which may disrupt their deleterious effects on the wound bed and microenvironment. By improving the wound microenvironment through the use of definitive proteases, these novel wound dressings may help transition wounds into the subsequent phase of healing.