Significance of in situ quantitative membrane property-morphology relation (QmPMR) analysis.

Deformation of the cell membrane is well understood from the viewpoint of protein interactions and free energy balance. However, the various dynamic properties of the membrane, such as lipid packing and hydrophobicity, and their relationship with cell membrane deformation are unknown. Therefore, the deformation of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and oleic acid (OA) giant unilamellar vesicles (GUVs) was induced by heating and cooling cycles, and time-lapse analysis was conducted based on the membrane hydrophobicity and physical parameters of "single-parent" and "daughter" vesicles. Fluorescence ratiometric analysis by simultaneous dual-wavelength detection revealed the variation of different hydrophilic GUVs and enabled inferences of the "daughter" vesicle composition and the "parent" membrane's local composition during deformation; the "daughter" vesicle composition of OA was lower than that of the "parents", and lateral movement of OA was the primary contributor to the formation of the "daughter" vesicles. Thus, our findings and the newly developed methodology, named in situ quantitative membrane property-morphology relation (QmPMR) analysis, would provide new insights into cell deformation and accelerate research on both deformation and its related events, such as budding and birthing.

Probiotic containing Lactobacillus reuteri DSM 17648 as an adjunct treatment for Helicobacter pylori infection: A randomized, double-blind, placebo-controlled trial.

BACKGROUND: Despite multiple therapy regimens, the decline in the Helicobacter pylori eradication rate poses a significant challenge to the medical community. Adding Lactobacillus reuteri probiotic as an adjunct treatment has shown some promising results. This study aims to investigate the efficacy of Lactobacillus reuteri DSM 17648 in H. pylori eradication and its effect in ameliorating gastrointestinal symptoms and adverse treatment effects. MATERIALS AND METHODS: This randomized, double-blinded, placebo-controlled trial involved treatment-naïve H. pylori-positive patients. Ninety patients received standard triple therapy for 2 weeks before receiving either a probiotic or placebo for 4 weeks. The posttreatment eradication rate was assessed via a 14 C urea breath test in Week 8. The Gastrointestinal Symptom Rating Scale (GSRS) questionnaire and an interview on treatment adverse effects were conducted during this study. RESULTS: The eradication rate was higher in the probiotic group than in the placebo group, with a 22.2% difference in the intention-to-treat analysis (91.1% vs. 68.9%; p = 0.007) and 24.3% difference in the per-protocol analysis (93.2% vs. 68.9%; p = 0.007). The probiotic group showed significant pre- to post-treatment reductions in indigestion, constipation, abdominal pain, and total GSRS scores. The probiotic group showed significantly greater reductions in GSRS scores than the placebo group: indigestion (4.34 ± 5.00 vs. 1.78 ± 5.64; p = 0.026), abdominal pain (2.64 ± 2.88 vs. 0.89 ± 3.11; p = 0.007), constipation (2.34 ± 3.91 vs. 0.64 ± 2.92; p = 0.023), and total score (12.41 ± 12.19 vs. 4.24 ± 13.72; p = 0.004). The probiotic group reported significantly fewer adverse headache (0% vs. 15.6%; p = 0.012) and abdominal pain (0% vs. 13.3%; p = 0.026) effects. CONCLUSIONS: There was a significant increase in H. pylori eradication rate and attenuation of symptoms and adverse treatment effects when L. reuteri was given as an adjunct treatment.

Implications of SARS-CoV-2 on current and future operation and management of wastewater systems.

While researchers have acknowledged the potential role of environmental scientists, engineers, and industrial hygienists during this pandemic, the role of the water utility professional is often overlooked. The wastewater sector is critical to public health protection and employs collection and treatment system workers who perform tasks with high potential for exposures to biological agents. While various technical guidances and reports have initially provided direction to the water sector, the rapidly growing body of research publications necessitates the constant review of these papers and data synthesis. This paper presents the latest findings and highlights their implications from a water and wastewater utility operation and management perspective. PRACTITIONER POINTS: Extrapolation from SARS-CoV-1 and MERS-CoV, as well as other surrogates, has helped predicting SARS-CoV-2 behavior and risk management. Data from treated wastewater effluent suggest that current processes are sufficient for SARS-CoV-2 control. Scientific evidence supports the possibility of fecal-oral transmission for SARS-CoV-2. Limited evidence supports the potential survival of infective SARS-CoV-2 on surfaces and in aerosols and the efficacy of control measures at reducing transmission. Protective practices and PPE can protect workers from SARS-CoV-2 and other pathogens found in wastewater.

A phase II trial of single oral FGF inhibitor, AZD4547, as second or third line therapy in malignant pleural mesothelioma.

OBJECTIVES: Currently, there is no optimal salvage therapy for patients with malignant pleural mesothelioma (MPM) who relapse after treatment with first-line chemotherapy. In line with the strong preclinical rationale for targeting fibroblast growth factor receptor (FGFR) signalling in malignant mesothelioma, we conducted a phase II study assessing the efficacy of AZD4547, an oral tyrosine multi-kinase FGFR 1-3 inhibitor, as a second or third-line treatment. MATERIALS AND METHODS: We conducted a single-center, open-label, single-arm phase II study of AZD4547 in eligible patients with confirmed, measurable MPM and radiological progression after first or second-line systemic chemotherapy. Patients received continuous, twice-daily oral AZD4547 on a 3-weekly cycle. The primary end point was 6-month progression free survival (PFS6). Response was assessed with CT scan every 6 weeks according to the modified RECIST criteria for mesothelioma (mRECIST) and toxicities were also assessed. The study used a Simon's two-stage design: 26 patients would be recruited to the first stage and more than 7 (27 %) of 26 patients were required to achieve PFS6 to continue to stage two, for a potential total cohort of 55 patients. RESULTS: 3 of 24 patients (12 %) were progression-free at 6 months. Hence, the study fulfilled stopping criteria regardless of further recruitment and warranted discontinuation. The most common toxicities (across all grades) were hyperphosphatemia, xerostomia, mucositis, retinopathy, dysgeusia, and fatigue. Maximum toxicities were grade 2 or below for all patients across all cycles. There was no association between tumour BAP1 protein loss and clinical outcomes. CONCLUSIONS: The FGFR 1-3 inhibitor AZD4547 did not demonstrate efficacy in patients with MPM who had progressed after first line treatment with platinum-based chemotherapy.

A Workflow to Improve Variant Calling Accuracy in Molecular Barcoded Sequencing Reads.

Multiplexed molecular barcoded amplicon sequencing has been previously demonstrated to improve the sensitivity of low-frequency variant detection. Molecular barcoded reads can be used to identify and correct amplification biases introduced during library preparation and sequencing errors. We propose a generic workflow to improve variant calling accuracy that takes advantage of molecular barcoded sequencing reads by applying a base score correction method to duplicate or overlapping read pairs across the targeted panel. The workflow is able to reduce the false-positive rate of variant calls in homopolymer and repetitive regions where the sequencer commonly encounters phasing errors interfering with base calling. The analysis was focused on three specific regions within a custom QIAseq targeted DNA (Qiagen, USA) of 220 genes observed to have a high false-positive rate in a clinical validation study. Uncorrected and corrected datasets were compared at targeted regions to reference calls from NIST and exome data from a reference laboratory (Macrogen, USA). Base correction removed all false positives identified without the correction method and retained the true positive call in the dataset. We have shown that our workflow incorporating base correction of molecular barcoded sequencing data can be applied to germline sequencing to improve variant calling accuracy in genomic regions with certain repetitive sequence motifs.

WASH activities at two Ebola treatment units in Sierra Leone.

PURPOSE: The 2014 outbreak of Ebola virus disease (EVD) in West Africa was the largest in history. Starting in September 2014, International Medical Corps (IMC) operated five Ebola treatment units (ETUs) in Sierra Leone and Liberia. This paper explores how future infectious disease outbreak facilities in resource-limited settings can be planned, organized, and managed by analyzing data collected on water, sanitation, and hygiene (WASH) and infection prevention control (IPC) protocols. DESIGN/METHODOLOGY/APPROACH: We conducted a retrospective cohort study by analyzing WASH/IPC activity data routinely recorded on paper forms or white boards at ETUs during the outbreak and later merged into a database from two IMC-run ETUs in Sierra Leone between December 2014 and December 2015. FINDINGS: The IMC WASH/IPC database contains data from over 369 days. Our results highlight parameters key to designing and maintaining an ETU. High concentration chlorine solution usage was highly correlated with both daily patient occupancy and high-risk zone staff entries; low concentration chlorine usage was less well explained by these measures. There is high demand for laundering and disinfecting of personal protective equipment (PPE) on a daily basis and approximately 1 (0-4) piece of PPE is damaged each day. RESEARCH LIMITATIONS/IMPLICATIONS: Lack of standardization in the type and format of data collected at ETUs made constructing the WASH/IPC database difficult. However, the data presented here may help inform humanitarian response operations in future epidemics.

Forward osmosis for oily wastewater reclamation: Multi-charged oxalic acid complexes as draw solutes.

Forward osmosis (FO) has demonstrated its merits in hybrid FO seawater desalination. However, FO may have a potential for other applications if suitable draw solutes are available. In this study, a series of novel draw solutes based on oxalic acid (OA)-transitional metal complexes are presented. Influential factors of FO performance have been systematically investigated by varying the transitional metals, cations of the complex draw solutes as well as the experimental conditions. Compared to NaCl and other recently synthesized draw solutes, the OA complexes show superior FO performance in terms of high water fluxes up to 27.5 and 89.1 LMH under the respective FO and PRO (pressure retarded osmosis) modes, both with negligible reverse solute fluxes. The features of octahedral geometry, abundant hydrophilic groups and ionic species are crucial for the OA complexes as appropriate draw solutes with satisfactory FO performance. Among the synthesized OA complexes, the ammonium salt of chromic complex (NH4-Cr-OA) outperforms others due to the presence of more ionic species in its complex system. NH4-Cr-OA also performs better than the typical NaCl draw solute in FO oily wastewater treatment with higher water recovery and negligible reverse fluxes. Dilute solutions of OA complexes have been reconcentrated through membrane distillation (MD) and reused to new round of FO processes. The OA complexes have demonstrated their suitability and superiority as a novel class of draw solutes for the FO process in this study.

An Effective Design of Electrically Conducting Thin-Film Composite (TFC) Membranes for Bio and Organic Fouling Control in Forward Osmosis (FO).

The organic foulants and bacteria in secondary wastewater treatment can seriously impair the membrane performance in a water treatment plant. The embedded electrode approach using an externally applied potential to repel organic foulants and inhibit bacterial adhesion can effectively reduce the frequency of membrane replacement. Electrode embedment in membranes is often carried out by dispensing a conductor (e.g., carbon nanotubes, or CNTs) in the membrane substrate, which gives rise to two problems: the leaching-out of the conductor and a percolation-limited membrane conductivity that results in an added energy cost. This study presents a facile method for the embedment of a continuous electrode in thin-film composite (TFC) forward osmosis (FO) membranes. Specifically, a conducting porous carbon paper is used as the understructure for the formation of a membrane substrate by the classical phase inversion process. The carbon paper and the membrane substrate polymer form an interpenetrating structure with good stability and low electrical resistance (only about 1Ω/□). The membrane-electrode assembly was deployed as the cathode of an electrochemical cell, and showed good resistance to organic and microbial fouling with the imposition of a 2.0 V DC voltage. The carbon paper-based FO TFC membranes also possess good mechanical stability for practical use.

Paediatric emergency department referrals from primary care.

Background Over the last decade, paediatric referrals from general practitioners (GPs) to the emergency department (ED) have increased by 60% in Australia. Objective To investigate the characteristics of Victorian children referred by GPs to the ED with lower-urgency conditions. Method Data were collected from four hospital EDs in Victoria, May-November 2014. Parents attending the ED with their child triaged as lower urgency were surveyed. Descriptive, frequency, and bivariate analyses were performed. Results Of the 1150 responses, 28% (320) visited their GP before attending ED. Of these 66% (212), were referred by their GP. A greater proportion with injury than illness (84% vs 59%; P<0.0001) was referred to the ED if they had first visited their GP. Conclusion Motivations of GPs to send lower-urgency injured and ill children to ED are not well understood. The high number of referrals from GPs to the ED for lower urgency conditions suggests attention by policy makers and health professionals must be paid to the current patterns of care of children in general practice. What is known about the topic? Paediatric referrals in Australia from GPs to EDs have increased in the last decade, along with the absolute number of children in Victoria presenting to the ED. What does this paper add? A significant number of children (66%) who attend the GP before visiting the ED are referred to the ED for their lower urgency condition. What are the implications for practitioners? It may be appropriate for GPs to be further supported to manage lower urgency conditions, through better resources or education.

Regular source of primary care and emergency department use of children in Victoria.

AIM: The aim of this paper was to study the prevalence of a regular source of primary care for Victorian children attending one of four emergency departments (EDs) and to determine associated characteristics, including ED use. METHODS: Responses were collected via an electronic survey from parents attending EDs with their child (≤9 years of age) for a lower-urgency condition. Single, multiple choice, and Likert scale responses were analysed using bivariate and logistic regression tests. RESULTS: Of the 1146 parents who provided responses, 80% stated their child has a regular source of primary care. Of these, care is mostly received by a general practitioner (GP) (95%) in GP group practices (71%). Approximately 20% have changed where their child receives primary care in the last year. No associations were observed between having a regular source of primary care and frequency of ED attendance in the past 12 months, although parents whose child did not have a regular source of primary care were more likely to view the ED as a more convenient place to receive care than the primary care provider (39% without regular source vs. 18% with regular source; P < 0.0001). Children were less likely to have a regular source of primary care if their parents were younger, had a lower household income, lower education, and were visiting a hospital in a lower socio-economic indexes for areas rank. CONCLUSIONS: Policy options to improve continuity of care for children may require investigation. Increasing the prevalence of regular source of primary care for children may in turn reduce ED visits.

Reducing the incidence of oxyhaemoglobin desaturation during rapid sequence intubation in a paediatric emergency department.

OBJECTIVES: Rapid sequence intubation (RSI) is the standard for definitive airway management in emergency medicine. In a video-based study of RSI in a paediatric emergency department (ED), we reported a high degree of process variation and frequent adverse effects, including oxyhaemoglobin desaturation (SpO2<90%). This report describes a multidisciplinary initiative to improve the performance and safety of RSI in a paediatric ED. METHODS: We conducted a local improvement initiative in a high-volume academic paediatric ED. We simultaneously tested: (1) an RSI checklist, (2) a pilot/copilot model for checklist execution, (3) the use of a video laryngoscope and (4) the restriction of laryngoscopy to specific providers. Data were collected primarily by video review during the testing period and the historical period (2009-2010, baseline). We generated statistical process control charts (G-charts) to measure change in the performance of six key processes, attempt failure and the occurrence of oxyhaemoglobin desaturation during RSI. We iteratively revised the four interventions through multiple plan-do-study-act cycles within the Model for Improvement. RESULTS: There were 75 cases of RSI during the testing period (July 2012-September 2013). Special cause variation occurred on the G-charts for three of six key processes, attempt failure and desaturation, indicating significant improvement. The frequency of desaturation was 50% lower in the testing period than the historical (16% vs 33%). When all six key processes were performed, only 6% of patients experienced desaturation. CONCLUSIONS: Following the simultaneous introduction of four interventions in a paediatric ED, RSI was performed more reliably, successfully and safely.

Calcium influx affects intracellular transport and membrane repair following nanosecond pulsed electric field exposure.

The cellular response to subtle membrane damage following exposure to nanosecond pulsed electric fields (nsPEF) is not well understood. Recent work has shown that when cells are exposed to nsPEF, ion permeable nanopores (<2 nm) are created in the plasma membrane in contrast to larger diameter pores (>2 nm) created by longer micro- and millisecond duration pulses. Nanoporation of the plasma membrane by nsPEF has been shown to cause a transient increase in intracellular calcium concentration within milliseconds after exposure. Our research objective is to determine the impact of nsPEF on calcium-dependent structural and repair systems in mammalian cells. Chinese hamster ovary (CHO-K1) cells were exposed in the presence and absence of calcium ions in the outside buffer to either 1 or 20, 600-ns duration electrical pulses at 16.2 kV/cm, and pore size was determined using propidium iodide and calcium green. Membrane organization was observed with morphological changes and increases in FM1-43 fluorescence. Migration of lysosomes, implicated in membrane repair, was followed using confocal microscopy of red fluorescent protein-tagged LAMP1. Microtubule structure was imaged using mEmerald-tubulin. We found that at high 600-ns PEF dosage, calcium-induced membrane restructuring and microtubule depolymerization coincide with interruption of membrane repair via lysosomal exocytosis.

Systemic but not topical TRAIL-expressing mesenchymal stem cells reduce tumour growth in malignant mesothelioma.

Malignant pleural mesothelioma is a rare but devastating cancer of the pleural lining with no effective treatment. The tumour is often diffusely spread throughout the chest cavity, making surgical resection difficult, while systemic chemotherapy offers limited benefit. Bone marrow-derived mesenchymal stem cells (MSCs) home to and incorporate into tumour stroma, making them good candidates to deliver anticancer therapies. Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) is a pro-apoptotic molecule that selectively induces apoptosis in cancer cells, leaving healthy cells unaffected. We hypothesised that human MSCs expressing TRAIL (MSCTRAIL) would home to an in vivo model of malignant pleural mesothelioma and reduce tumour growth. Human MSCs transduced with a lentiviral vector encoding TRAIL were shown in vitro to kill multiple malignant mesothelioma cell lines as predicted by sensitivity to recombinant TRAIL (rTRAIL). In vivo MSC homing was delineated using dual fluorescence and bioluminescent imaging, and we observed that higher levels of MSC engraftment occur after intravenous delivery compared with intrapleural delivery of MSCs. Finally, we show that intravenous delivery of MSCTRAIL results in a reduction in malignant pleural mesothelioma tumour growth in vivo via an increase in tumour cell apoptosis.

Dry skin in the winter is related to the ceramide profile in the stratum corneum and can be improved by treatment with a Eucalyptus extract.

BACKGROUND: Dry skin in the winter has been reported to involve scaling, defects in water holding and barrier functions, and decreased ceramide (CER) levels in the stratum corneum (SC). We previously reported that a Eucalyptus extract promotes CER synthesis in cultured keratinocytes and accelerates the recovery of hydration in a barrier-disrupted model of human skin. AIMS: One of the objectives was to examine the CER profile and its contribution to the relief of dry skin. The other objective was to assess the efficacy of a Eucalyptus extract to treat dry skin. PATIENTS/METHODS: Twenty subjects with dry skin on their legs were assessed and their CER profiles were analyzed using tape-stripping. A moisturizer with a Eucalyptus extract was assessed for its effects on dry skin using a leg regression methodology comprising 28 days of treatment and 14 days of regression. RESULTS: Indicators of dry skin conditions (conductance, dryness, roughness, and scaliness) strongly correlated with the level of CER, CER [NP], and CER[NH]. Treatment with the Eucalyptus extract significantly improved conductance (3 days after regression) and transepidermal water loss (14 days after regression) compared with the placebo. After 28 days of treatment with the Eucalyptus extract, the level of CER in the SC did not increase, but CER [NP] did increase. CONCLUSIONS: These results suggest that not only the level of CER, but also specific CER species strongly contribute to dry skin relief and products that increase those are useful to improve dry skin conditions.

In situ simulation: detection of safety threats and teamwork training in a high risk emergency department.

OBJECTIVE: Implement and demonstrate feasibility of in situ simulations to identify latent safety threats (LSTs) at a higher rate than lab-based training, and reinforce teamwork training in a paediatric emergency department (ED). METHODS: Multidisciplinary healthcare providers responded to critical simulated patients in an urban ED during all shifts. Unannounced in situ simulations were limited to 10 min of simulation and 10 min of debriefing, and were video recorded. A standardised debriefing template was used to assess LSTs. The primary outcome measure was the number and type of LSTs identified during the simulations. Secondary measures included: participants' assessment of impact on patient care and value to participants. Blinded video review using a modified Anaesthetists Non-Technical Skills scale was used to assess team behaviours. RESULTS: 218 healthcare providers responded to 90 in situ simulations conducted over 1 year. A total of 73 LSTs were identified; a rate of one every 1.2 simulations performed. In situ simulations were cancelled at a rate of 28% initially, but the cancellation rate decreased as training matured. Examples of threats identified include malfunctioning equipment and knowledge gaps concerning role responsibilities. 78% of participants rated the simulations as extremely valuable or valuable, while only 5% rated the simulation as having little or no value. Of those responding to a postsimulation survey, 77% reported little or no clinical impact. Video recordings did not indicate changes in non-technical skills during this time. CONCLUSIONS: In situ simulation is a practical method for the detection of LSTs and to reinforce team training behaviours. Embedding in situ simulation as a routine expectation positively affected operations and the safety climate in a high risk clinical setting.