Comparison of methods for sampling and detection of carbapenemase-producing organisms in clinical handwash basin drains in health care.

There are currently no standardized methods for the sampling and testing of clinical handwash basin (HWB) samples for the detection of carbapenemase-producing organisms (CPOs). Methods used for sampling (drain aspirate vs swab from top of drain) and detection of CPOs in clinical HWB drains in two different healthcare settings, one which was dealing with a hospital-wide CPO outbreak (Hospital A) and another with no reported outbreaks (Hospital B), were compared. Drain aspirates and swabs from HWB drains were tested using multiplex polymerase chain reaction (PCR) together with culture-based methods. No significant difference in detection of CPOs was found between drain aspirate or swab methods of sampling. Direct PCR on samples detected significantly more carbapenemase genes than culture on CARBA agar (P<0.0001 and 0.0045, respectively). A higher percentage of HWB drains were positive in Hospital A both by culture and by direct PCR, and a significantly higher number of carbapenemase genes were detected in HWB drain aspirates at Hospital A, both by PCR and by culture (P=0.014 and 0.0071, respectively). There was high correlation between drain swab positivity by PCR and culture in Hospital A (91%) compared with Hospital B (33%). No difference in drain contamination rates was found when HWBs with a rear drain were compared with HWBs with the drain directly below the tap. Colonization of HWBs at the top of the drain may be related to risk of cross-transmission of CPOs from the healthcare environment to patients.

Candidacy and long-term outcomes of subcutaneous implantable cardioverter-defibrillators in current practice in patients with hypertrophic cardiomyopathy.

BACKGROUND: In patients with Hypertrophic Cardiomyopathy (HCM) S-ICD is usually the preferred option as pacing is generally not indicated. However, limited data are available on its current practice adoption and long-term follow-up. METHODS: Consecutive HCM patients with S-ICD implanted between 2013 and 2021 in 3 international centers were enrolled in this observational study. Baseline, procedural and follow-up data were regularly collected. Efficacy and safety were compared with a cohort of HCM patients implanted with a tv-ICD. RESULTS: Seventy patients (64% males) were implanted with S-ICD at 41 ± 15 years, whereas 168 patients with tv-ICD at 49 ± 16 years. For S-ICD patients, mean ESC SCD risk score was 4,5 ± 1.9%: 25 (40%) at low-risk, 17 (27%) at intermediate and 20 (33%) at high-risk. Patients were followed-up for 5.1 ± 2.3 years. Two patients (0.6 per 100-person-years, vs 0.4 per 100 person-years with tv-ICD, p = 0.45) received an appropriate shock on VF, 17 (24%) were diagnosed with de-novo AF. Inappropriate shocks occurred in 4 patients (1.2 per 100-person-years, vs 0.9 per 100 person-years with tv-ICD, p = 0.74), all before Smart-Pass algorithm implementation. Four patients experienced device-related adverse events (1.2 per 100-person-years, vs 1 per 100 person-years with tv-ICD, p = 0.35%). CONCLUSIONS: S-ICDs were often implanted in patients with an overall low-intermediate ESC SCD risk, reflecting both the inclusion of additional risk markers and a lower decision threshold. S-ICDs in HCM patients followed for over 5 years showed to be effective in conversion of VF and safe. Greater scrutiny may be required to avoid overtreatment in patients with milder risk profiles.

Assessment of mould remediation in a healthcare setting following extensive flooding.

BACKGROUND: A new hospital building was close to completion when a large pipe carrying clean water broke, causing extensive flooding. AIM: To determine the flood-associated fungal risk to susceptible patients who would use that building. METHODS: Though standard flood remediation by the builders was relatively straightforward, there was no model for specialist assessment of patient risk due to the flood-associated mould growth. As levels of background airborne fungal spores can be expected to vary significantly over time, we could not use absolute levels to indicate either an excess of airborne fungal spores or successful remediation. Therefore it was decided to use weekly settle plates, exposed at the same time in flooded (test) and equivalent non-flooded (control) areas to compensate for variations in background levels. Flood-related risk was estimated by the ratio between fungal colonies on the test and control sets of settle plates, rather than absolute number. FINDINGS: Whereas the physical flood remediation, including the use of 'anti-fungal' treatments, was completed in three weeks post flooding, fungal contamination in flooded areas took 38 weeks to return to control levels and remained so for a further six weeks of observation. CONCLUSION: By the use of this method, we were able to assure the absence of flood-associated fungal risk to susceptible patients who would use that building. We recommend that infection prevention and control teams consider using this approach should they be faced with similar situations.

Whole-genome sequencing links cases dispersed in time, place, and person while supporting healthcare worker management in an outbreak of Panton-Valentine leucocidin meticillin-resistant Staphylococcus aureus; and a review of literature.

This is a report on an outbreak of Panton-Valentine leucocidin-producing meticillin-resistant Staphylococcus aureus (PVL-MRSA) in an intensive care unit (ICU) during the COVID-19 pandemic that affected seven patients and a member of staff. Six patients were infected over a period of ten months on ICU by the same strain of PVL-MRSA, and a historic case identified outside of the ICU. All cases were linked to a healthcare worker (HCW) who was colonized with the organism. Failed topical decolonization therapy, without systemic antibiotic therapy, resulted in ongoing transmission and one preventable acquisition of PVL-MRSA. The outbreak identifies the support that may be needed for HCWs implicated in outbreaks. It also demonstrates the role of whole-genome sequencing in identifying dispersed and historic cases related to the outbreak, which in turn aids decision-making in outbreak management and HCW support. This report also includes a review of literature of PVL-MRSA-associated outbreaks in healthcare and highlights the need for review of current national guidance in the management of HCWs' decolonization regimen and return-to-work recommendations in such outbreaks.

The combined measurement of synovial markers in the diagnosis of periprosthetic joint infection.

INTRODUCTION: Currently there is no single test for periprosthetic joint infection (PJI) that demonstrates an acceptable level of sensitivity. The aim of this pilot study was to identify a combination of biomarkers that could exclude periprosthetic infection prior to revision surgery in hip and knee arthroplasty. This would help in equivocal cases of PJI, such as those with low-virulence organisms, chronic low-grade infection or 'aseptic' loosening. Current research has focused on measuring the levels of biomarkers in the patient's synovial fluid, and these can be combined to improve accuracy. METHODS: We selected synovial white cell count (SF-WCC) and C-reactive protein (SF-CRP), as they demonstrated an acceptable level of sensitivity and specificity which could be measured using existing equipment. One hundred and sixty-one synovial fluid samples from 134 patients were collected and analysed prospectively using calculated cut-off values of 10mg/l for SF-CRP and 3,000×106/l for SF-WCC. Samples were deemed positive for infection when either of the SF-CRP or SF-WCC values were above the cut-off. RESULTS: The combined test demonstrated a sensitivity greater than 98.5% and specificity above 80% in all samples analysed or looked at separately for total hip replacement and total knee replacement infections. All ten aspirates taken from chronically infected joints tested positive. CONCLUSIONS: The combination of SF-CRP and SF-WCC measurement has been shown to have high sensitivity of over 99% in detecting both acute and chronic PJI in both hip and knee arthroplasty. This ability of the test to exclude infection with a high degree of certainty will help in preoperative planning of PJIs.

Unintended consequences of long-sleeved gowns in a critical care setting during the COVID-19 pandemic.

At the peak of the coronavirus disease 2019 (COVID-19) pandemic, hand hygiene audits indicated decreased compliance in a 12-bed critical care (CC) area with ventilated COVID-19 patients, where staff used personal protective equipment (PPE), including sessional use of long-sleeved gowns in accordance with the recommendations of Public Health England. There was also a cluster of three central venous catheter (CVC) infections along with increases in the number of patients from whom enteric Gram-negative bacteria (GNB) were isolated from sterile sites. Environmental sampling of near-patient surfaces and frequently touched sites demonstrated that 11.5% of areas were contaminated with enteric GNB in the COVID-19 CC area, compared with 2.6% and 2.7% in COVID-19 and non-COVID-19 general wards, respectively. Following a risk assessment, hospital policy was changed to replace long-sleeved gowns with short-sleeved gowns. The CC unit underwent enhanced cleaning with hypochlorite-based disinfectant and was resampled 8 days later. On resampling, no GNB were isolated from the CC unit. Following this change in PPE, hand hygiene compliance returned to baseline standards and no further CVC infections were identified. Staff reported a preference for short-sleeved gowns. No evidence currently exists that PPE beyond that recommended for pandemic influenza (respiratory protection plus standard PPE) adds to the protection of healthcare workers (HCWs) from severe acute respiratory syndrome coronavirus-2. Long-sleeved gowns prevent HCWs performing hand hygiene effectively. While it is imperative that HCWs are adequately protected, protection of patients from infection hazards is equally important. Further studies are necessary to establish risks from PPE to inform a review of current guidance.

Chyawanprash: A Traditional Indian Bioactive Health Supplement.

Chyawanprash (CP) is an Ayurvedic health supplement which is made up of a super-concentrated blend of nutrient-rich herbs and minerals. It is meant to restore drained reserves of life force (ojas) and to preserve strength, stamina, and vitality, while stalling the course of aging. Chyawanprash is formulated by processing around 50 medicinal herbs and their extracts, including the prime ingredient, Amla (Indian gooseberry), which is the world's richest source of vitamin C. Chyawanprash preparation involves preparing a decoction of herbs, followed by dried extract preparation, subsequent mixture with honey, and addition of aromatic herb powders (namely clove, cardamom, and cinnamon) as standard. The finished product has a fruit jam-like consistency, and a sweet, sour, and spicy flavor. Scientific exploration of CP is warranted to understand its therapeutic efficacy. Scattered information exploring the therapeutic potential of CP is available, and there is a need to assemble it. Thus, an effort was made to compile the scattered information from ancient Ayurvedic texts and treatises, along with ethnobotanical, ethnopharmacological, and scientifically validated literature, that highlight the role of CP in therapeutics. Citations relevant to the topic were screened.

NIR-light active hybrid nanoparticles for combined imaging and bimodal therapy of cancerous cells.

We report the synthesis of a multifunctional biocompatible theranostic nanoplatform consisting of a biodegradable PLGA matrix surface-functionalized with indocyanine green (ICG), a near-IR fluorescent dye, and co-loaded with superparamagnetic iron oxide nanoparticles (SPIONs) and the anticancer drug doxorubicin (DOXO). Combination of chemo- and photothermal therapeutic efficacy as well as magnetic resonance and optical fluorescence imaging performance were successfully tested in vitro on a tumoral cervical HeLa cell line. Magnetic in vitro guided targeting of these nanoplatforms was also proven. These nanoconstructs also enabled to monitor their in vivo biodistribution by fluorescence imaging in a mice model, which revealed their effective accumulation in the tumor and, unexpectedly, in the brain area. A lower presence of nanoplatforms was noted in the reticulo-endothelial system. The present observations suggest the nanoplatforms ability to possibly overcome the blood brain barrier. These results open up new possibilities to use our multifunctional nanoplatforms to treat brain-located diseases.

Efficient production of Hantaan and Puumala pseudovirions for viral tropism and neutralization studies.

Puumala (PUUV) and Hantaan (HTNV) viruses are hantaviruses within the family Bunyaviridae and associated with Hemorrhagic Fever with Renal Syndrome (HFRS) in humans. Little is known about how these viruses interact with host cells, though pathogenic hantaviruses interact with α(v)ß(3) integrin. To study host cell interactions and rapidly test the ability of antibodies to prevent infection, we produced HTNV and PUUV pseudovirions on a vesicular stomatitis virus (VSV) core. Similar to replication-competent hantaviruses, infection was low-pH-dependent. Despite broad cell tropism, several human T cell lines were poorly permissive to hantavirus pseudovirions, compared to VSV, indicating a relative block to infection at the level of entry. Stable expression of α(v)ß(3) integrin in SupT1 cells did not restore infectivity. Finally, the pseudovirion system provided a rapid, quantitative, and specific method to screen for neutralizing antibodies in immune sera.

WITHDRAWN. Universal neonatal hearing screening versus selective screening as part of the management of childhood deafness.

BACKGROUND: The principal factors that decide how deafness affects a child's development are the degree of hearing impairment and the age at which it is diagnosed. A number of factors are thought to increase the risk of hearing impairment: low birth weight, prematurity, perinatal hypoxia and jaundice, among others. The high incidence of deafness in children without risk factors and the introduction of simple new screening tests of high sensitivity and specificity have led many prestigious bodies to recommend universal early detection programmes for deafness rather than screening targeted only at high-risk groups. OBJECTIVES: To compare the long-term effectiveness of a universal neonatal screening and early treatment programme for hearing impairment with: a) screening and treatment only of high-risk neonates and b) opportunistic screening and treatment. SEARCH STRATEGY: Databases searched included MEDLINE (1966 to 2006), EMBASE (1974 to 2006), the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library Issue 2, 2006) and registers of health technology assessment agencies as well as registers of clinical guidelines. The date of the last search was June 2006. SELECTION CRITERIA: Randomised controlled trials comparing universal neonatal screening with either high-risk screening or opportunistic screening for hearing impairment. DATA COLLECTION AND ANALYSIS: Two review authors independently screened the search results to identify suitable trials. MAIN RESULTS: No studies were identified which fulfilled the inclusion criteria. AUTHORS' CONCLUSIONS: The long-term effectiveness of universal newborn hearing screening programmes has not been established to date. There is a need for controlled trials and before and after studies to address this issue further.

Metabolic syndrome attenuates effect of chronic kidney disease on prevalence of coronary disease in men referred for stress imaging study.

BACKGROUND: Metabolic syndrome (MS) and chronic kidney disease (CKD) are both strongly associated with coronary artery disease (CAD). Components of MS also cause CKD. The incremental effect of CKD on CAD prevalence in MS patients referred for stress imaging studies is unknown. METHODS: From January to December 2005, consecutive subjects referred for a stress imaging study were prospectively enrolled. CAD was defined as fixed or reversible defects on nuclear imaging and as resting or stress-induced wall motion abnormalities on echocardiography. MS was defined using NCEP-ATP III criteria. CKD definition was based on calculated glomerular filtration rate. The independent effect of CKD on stress results was assessed using multiple variable logistic regression. Stepwise model selection was used for variable reduction, and areas under the receiver operating characteristic curves (ROC) were calculated. RESULTS: Of 1,122 patients enrolled (mean age 61.4 years, 97% male), 535 (47%) had MS. Among MS patients, 156/535 (29%) had CKD while 116/587 (19%) subjects without MS had CKD. Subjects with CKD were older (p < 0.001) in subjects with and without MS. The presence of CKD affected prevalence of CAD in the non-MS group only, almost doubling it (20% vs. 38%, p < 0.001). Further, using the ordered nature of the 5 CKD stages, worsening severity of CKD had greater prevalence of CAD, in non-MS subjects only (p < 0.001). CONCLUSIONS: MS attenuates the effect of CKD on CAD prevalence, regardless of CKD severity. CKD almost doubles the prevalence of CAD in non-MS subjects. CKD severity is associated with greater CAD burden in the non-MS group.

Candida inferior vena cava filter infection and septic thrombophlebitis.

Candida infection following central venous catheterization (CVC) is a well known complication, most common infection being that caused by Candida albicans. Less common is infection with Candida glabrata. Candida septic thrombophlebitis of the central veins is an uncommon complication after CVC. We report the first case of inferior vena cava filter infection with C. glabrata following septic thrombophlebitis of the femoral veins and discuss management of this rare complication.

Molecular characterization of the Ran-binding zinc finger domain of Nup153.

The nuclear pore complex is the gateway for selective traffic between the nucleus and cytoplasm. To learn how building blocks of the pore can create specific docking sites for transport receptors and regulatory factors, we have studied a zinc finger module present in multiple copies within the nuclear pores of higher eukaryotes. All four zinc fingers of human Nup153 were found to bind the small GTPase Ran with dissociation constants ranging between 5 and 40 mum. In addition a fragment of Nup153 encompassing the four tandem zinc fingers was found to bind Ran with similar affinity. NMR structural studies revealed that a representative Nup153 zinc finger adopts the same zinc ribbon structure as the previously characterized Npl4 NZF module. Ran binding was mediated by a three-amino acid motif (Leu(13)/Val(14)/Asn(25)) located within the two zinc coordination loops. Nup153 ZnFs bound GDP and GTP forms of Ran with similar affinities, indicating that this interaction is not influenced by a nucleotide-dependent conformational switch. Taken together, these studies elucidate the Ran-binding interface on Nup153 and, more broadly, provide insight into the versatility of this zinc finger binding module.

Studying nuclear disassembly in vitro using Xenopus egg extract.

Xenopus egg extract provides an extremely powerful approach in the study of cell cycle regulated aspects of nuclear form and function. Each egg contains enough membrane and protein components to support multiple rounds of cell division. Remarkably, incubation of egg extract with DNA in the presence of an energy regeneration system is sufficient to induce formation of a nuclear envelope around DNA. In addition, these in vitro nuclei contain functional nuclear pore complexes, which form de novo and are capable of supporting nucleocytoplasmic transport. Mitotic entry can be induced by the addition of recombinant cyclin to an interphase extract. This initiates signaling that leads to disassembly of the nuclei. Thus, this cell-free system can be used to decipher events involved in mitotic remodeling of the nuclear envelope such as changes in nuclear pore permeability, dispersal of membrane, and disassembly of the lamina. Both general mechanisms and individual players required for orchestrating these events can be identified via biochemical manipulation of the egg extract. Here, we describe a procedure for the assembly and disassembly of in vitro nuclei, including the production of Xenopus egg extract and sperm chromatin DNA.