Petra Derrington

Innate Transcriptional Networks Activated in Bladder in Response to Uropathogenic Escherichia coli Drive Diverse Biological Pathways and Rapid Synthesis of IL-10 for Defense against Bacterial Urinary Tract Infection

Early transcriptional activation events that occur in bladder immediately following bacterial urinary tract infection (UTI) are not well defined. In this study, we describe the whole bladder transcriptome of uropathogenic Escherichia coli (UPEC) cystitis in mice using genome-wide expression profiling to define the transcriptome of innate immune activation stemming from UPEC colonization of the bladder. Bladder RNA from female C57BL/6 mice, analyzed using 1.0 ST-Affymetrix microarrays, revealed extensive activation of diverse sets of innate immune response genes, including those that encode multiple IL-family members, receptors, metabolic regulators, MAPK activators, and lymphocyte signaling molecules. These were among 1564 genes differentially regulated at 2 h postinfection, highlighting a rapid and broad innate immune response to bladder colonization. Integrative systems-level analyses using InnateDB (http://www.innatedb.com) bioinformatics and ingenuity pathway analysis identified multiple distinct biological pathways in the bladder transcriptome with extensive involvement of lymphocyte signaling, cell cycle alterations, cytoskeletal, and metabolic changes. A key regulator of IL activity identified in the transcriptome was IL-10, which was analyzed functionally to reveal marked exacerbation of cystitis in IL-10–deficient mice. Studies of clinical UTI revealed significantly elevated urinary IL-10 in patients with UPEC cystitis, indicating a role for IL-10 in the innate response to human UTI. The whole bladder transcriptome presented in this work provides new insight into the diversity of innate factors that determine UTI on a genome-wide scale and will be valuable for further data mining. Identification of protective roles for other elements in the transcriptome will provide critical new insight into the complex cascade of events that underpin UTI.

An assessment of high touch object cleaning thoroughness using a fluorescent marker in two Australian hospitals

Abstract Objective We needed to better understand the usefulness of different methods of monitoring and achieving sustained improvement in cleaning. Common elements of successful international approaches include a covert nature, the use of a method to visually identify and highlight deficiencies in the cleaning of high touch objects (HTOs) and the provision of feedback and education before re-evaluation. The specific purpose of this study was to evaluate fluorescent marking, education and feedback for assessing and improving HTO cleaning in a typical Australian inpatient hospital setting. Methods A three-phase, prospective study was conducted in two acute care hospitals over 17 weeks. For each phase, in a set of 37 specific single-inpatient rooms, seven predefined HTOs were marked with a liquid isopropyl alcohol and optical brightener formulation targeting material solution containing a fluorescent marker (FM), known as DAZO ® , designed specifically for the purpose of evaluating surface cleaning. In each hospital we targeted rooms located in the four wards with the greatest de novo multidrug resistant organism burden. Forty-eight hours after applying the FM we used a black-light to visualise the marks presence or removal. In phase 1 only, HTOs were swabbed before marking. Also in the first phase only and immediately following the initial assessment, eachHTOwas cleaned, remarked and re-assessed at 48 h. Between phases 1 and 2, investigators provided results to environmental services (EVS) leadership and staff. Education was provided to EVS staff after phase 1 only. Results A total of 986 marks were evaluated. The cleaning scores for individual HTOs in phases 1–3 ranged from 9.4 to 77.8%, 10.8 to 93% and 13.5 to 67.7% respectively. In phase 3, three HTOs scored lower than in phase 1. The mean overall cleaning scores for phases 1–3 were 34%, 53% and 41% respectively. Conclusions The FM was useful to assess HTO cleaning thoroughness. It facilitated relevant feedback and education and motivated staff to strive for continual improvements in environmental cleaning. Without on-going education, preliminary improvements were unsustained. However, investigators better understood flaws in cleaning and policy/procedure conflicts.

Predicting influenza A and 2009 H1N1 influenza in patients admitted to hospital with acute respiratory illness

Objective To create a clinical decision tool for suspected influenza A (including 2009 H1N1) to facilitate treatment and isolation decisions for patients admitted to hospital with an acute respiratory illness from the emergency department (ED) during a 2009 H1N1 pandemic. Methods Cross-sectional study conducted in two hospitals in Queensland, Australia. All patients admitted to hospital from the ED between 24 May and 16 August 2009 with an acute respiratory illness were included. All had nasal and throat swabs taken. Data were collected from clinical chart review regarding clinical symptoms, co-morbidities, examination findings, pathology and radiology results. Influenza A status was detected by reverse transcription–PCR assay. Univariate and multivariate regression analyses were performed to identify independent predictors of influenza A status. Results 346 consecutive patients were identified, of which 106 were positive for 2009 H1N1 influenza; an additional 11 patients were positive for other influenza A viruses. Independent clinical predictors (with points allocated using weighted scoring) for all types of influenza A in patients admitted with acute respiratory illness were: age 18–64 years (2 points); history of fever (2); cough (1); normal level of consciousness (2); C-reactive protein >5 and ≤100 mg/l (2) and normal leucocyte count (1). A clinical score of 5 (presence of two or three predictors) gave a sensitivity of 93% (95% CI 87% to 96%), specificity of 36% (95% CI 30% to 42%), resulting in a negative-predictive value of 91% (95% CI 83% to 95%). Conclusion A clinical prediction tool was developed that may be able to assist in making appropriate isolation decisions during future 2009 H1N1 outbreaks.

Protein-based profiling of the immune response to uropathogenic Escherichia coli in adult patients immediately following hospital admission for acute cystitis

Urinary tract infections (UTIs) caused by uropathogenic Escherichia coli (UPEC) are common infections in humans. Despite the substantial healthcare cost represented by these infections, the human immune response associated with the infection immediately following the onset of symptoms in patients remains largely undefined. We performed a prospective study aimed at defining the milieu of urinary cytokines in adult inpatients in the 24-48 h period immediately following hospital admission for acute cystitis due to UPEC. Urine samples, analyzed using 27-target multiplex protein assays, were used to generate immune profiles for patients and compared to age- and gender-matched healthy controls. The levels of multiple pro-inflammatory cytokines were significantly elevated in urine as a result of infection, an observation consistent with prior findings in murine models and clinical literature. We also identified significant responses for several novel factors not previously associated with the human response to UTI, including Interleukin (IL)-4, IL-7, IL-9, IL-17A, eotaxin, Granulocyte-macrophage colony-stimulating factor (GM-CSF) and several growth factors. These data establish crucial parallels between the human immune response to UPEC and murine model UTI studies, and emphasize the complex but poorly defined nature of the human immune response to UPEC, particularly in the immediate period following the onset of symptoms for acute cystitis.

SCEDOSPORIUM APIOSPERMUM PERITONITIS IN A PATIENT UNDERGOING PERITONEAL DIALYSIS

To the Editor: Renal interstitial fibrosis (RIF) plays a most important role in the progression of renal diseases to end-stage renal failure. Unilateral ureteral obstruction (UUO) model was used extensively as a model of progressive RIF, accompanied by rapid parenchymal deterioration. All-trans retinoic acid (ATRA), a natural derivative of vitamin A, plays an important role in the prevention of fibrosis. However, there was a rare report investigating the effect of ATRA on RIF. Interestingly, Kishimoto et al. performed an investigation on the association of ATRA treatment with RIF progression in UUO model mice, and found that a significant improvement was observed in histological and immunological findings in the ATRA treatment group, including macrophage infiltration and improved expression of monocyte chemotactic protein-1, transforming growth factor-b1 (TGF-b1), a-smooth muscle actin (a-SMA) and collagen I compared with the UUO model group. They drew a conclusion that ATRA treatment was not only an effective prophylactic strategy, but also a therapeutic strategy for the treatment of progressive renal fibrosis in diseased kidneys. We also performed an investigation on the association of the ATRA treatment with RIF progression. The rats were divided into three groups: sham operation group (SHO), group of UUO model (GU) and UUO model group treated with ATRA (GA). The UUO model was established, and the rats in the GA group were treated with ATRA (Sigma, St. Louis, MO, USA; 15 mg/kg per day) in corn oil once daily by oral gavage, from the first day before operation. Rats in the GU and SHO groups were administrated saline. Rats from the three groups were killed 14 and 28 days after surgery, and the renal tissue was collected for the determination of histological and molecular biology. In our study, we found that ATRA treatment could significantly reduce the expressions of TGF-b1, a-SMA, collagen IV and fibronectin. Our results were similar with those from Kishimoto et al. So, ATRA might become a useful therapeutic strategy for the treatment of progressive renal fibrosis. All-trans retinoic acid might play an important role in the administration of renal diseases in experimental studies. In our previous study, we found that ATRA could alleviate glomerulosclerosis lesion in glomerulosclerosis rats induced by adriamycin. ATRA might be a candidate drug for the treatment of renal diseases in a clinical setting. However, further investigation should be performed in experimental animal models and the effect of ATRA should be confirmed in the clinical practice.

Nothing to sneeze at! A study into intra‐operative contamination

Purpose:  We performed a prospective study of sneezes from orthopaedic registrars to assess the potential for intra‐operative contamination from a masked surgeon, and to determine if head position can alter the potential for contamination.

Outbreak of health care-associated Burkholderia cenocepacia bacteremia and infection attributed to contaminated sterile gel used for central line insertion under ultrasound guidance and other procedures

Background: We report an outbreak of Burkholderia cenocepacia bacteremia and infection in 11 patients predominately in intensive care units caused by contaminated ultrasound gel used in central line insertion and sterile procedures within 4 hospitals across Australia. Methods: Burkholderia cenocepacia was first identified in the blood culture of a patient from the intensive care unit at the Gold Coast University Hospital on March 26, 2017, with 3 subsequent cases identified by April 7, 2017. The outbreak response team commenced investigative measures. Results: The outbreak investigation identified the point source as contaminated gel packaged in sachets for use within the sterile ultrasound probe cover. In total, 11 patient isolates of B cenocepacia with the same multilocus sequence type were identified within 4 hospitals across Australia. This typing was the same as identified in the contaminated gel isolate with single nucleotide polymorphism‐based typing, demonstrating that all linked isolates clustered together. Conclusion: Arresting the national point‐source outbreak within multiple jurisdictions was critically reliant on a rapid, integrated, and coordinated response and the use of informal professional networks to first identify it. All institutions where the product is used should look back at Burkholderia sp blood culture isolates for speciation to ensure this outbreak is no larger than currently recognized given likely global distribution.

A pilot study comparing superficial wound swab, deep tissue biopsy and fine needle aspiration biopsy in identifying infecting organisms in foot ulcers due to diabetes

Background Current clinical practice widely regards deep tissue biopsy as the gold standard for identification of wound bacterial bio-burden. This study aims to establish whether fine needle aspiration biopsy (FNAB) is as accurate as deep tissue biopsy and therefore offers a more accurate, cheaper and suitable alternative to routinely used superficial swab in diabetic wounds of varying depth and severity.

Characterising a Ralstonia outbreak with a novel source

There are enormous challenges facing infection prevention and control in the 21st century. Countries across the world are confronted by ageing populations, restricted healthcare resources, demands for modern medicine and increasing antimicrobial resistance. Problem pathogens in the community are set to invade hospitals and those generated in hospitals are returned to the community. Continued consumption of antimicrobial agents is promoting and consolidating resistance to nearly all classes of drugs. Novel, known and resistant pathogens arising in one locality rapidly spread across the ‘global village’ courtesy of migration, conflict and international travel. We are facing unprecedented threats to the management of infection all over the world. This presentation will summarise the escalating challenges for infection control along with mention of current and future strategies for dealing with these. Despite the lack of evidence underpinning microbial transmission, basic hygiene practices remain the basis for protecting our patients. The science of infection prevention and control is set to become a key component in healthcare delivery. As we prepare for a world without antibiotics, research, education and policies on screening, isolation, cleaning and infection control have never been more important. Creative thinking, initiative and courage will be required for future management of infection in hospitals and communities everywhere.

Evaluation of immunochromatographic test kits for the detection of norovirus in faecal specimens

Background: The early detection of norovirus plays a vital role in the reduction of transmission and control of nosocomial viral gastroenteritis. Aim: This study evaluated the immunochromatographic test kits RIDAQUICK N1402, RIDAQUICK N1403, SD BIOLINE Norovirus and ImmunoCard STAT! Norovirus. There are no current publications evaluating RIDAQUICK N1402. Method: Eighty-six thawed faecal samples previously subjected to viral loading were tested with the kits and the cycle threshold value used as the standard reference. Twenty fresh faecal samples containing norovirus were also tested. Results: The sensitivity for RIDAQUICK N1403, RIDAQUICK N1402, SD BIOLINE Norovirus, ImmunoCard STAT! Norovirus, using the thawed samples was 46%, 58%, 32% and 47%, respectively. Specificity for RIDAQUICK N1403 was 96% whilst the other test kits obtained 100% specificity. Using the fresh samples, sensitivity was 70% and specificity was 100% across all kits. Discussion: Variances in sensitivities and specificities across test kits between the thawed and fresh samples indicate the importance of the storage of samples. Immunochromatographic test kits may be useful as a primary screening method for the detection of norovirus, however negative samples should be confirmed with more sensitive detection methods such as reverse transcriptase polymerase chain reaction (RT-PCR).