The fatigue-inducing effects of cancer and its therapy are characterized by decreased physical activity in the absence of any motivational deficit.

Although cancer and its therapy are well known to be associated with fatigue, the exact nature of cancer-related fatigue remains ill-defined. We previously reported that fatigue-like behavior induced independently by tumor growth and by the chemotherapeutic agent cisplatin is characterized by reduced voluntary wheel running and an intact motivation to expand effort for food rewards. The present set of experiments was initiated to characterize the functional consequences of fatigue induced by chemoradiotherapy in tumor-bearing mice and relate them to changes in the expression of genes coding for inflammation, mitochondria dynamics and metabolism. Two syngeneic murine models of cancer were selected for this purpose, a model of human papilloma virus-related head and neck cancer and a model of lung cancer. In both models, tumor-bearing mice were submitted to chemoradiotherapy to limit tumor progression. Two dimensions of fatigue were assessed, the physical dimension by changes in physical activity in mice trained to run in wheels and the motivational dimension by changes in the performance of mice trained to nose poke to obtain a food reward in a progressive ratio schedule of food reinforcement. Chemoradiotherapy reliably decreased wheel running activity but had no effect on performance in the progressive ratio in both murine models of cancer. These effects were the same for the two murine models of cancer and did not differ according to sex. Livers and brains were collected at the end of the experiments for qRT-PCR analysis of expression of genes coding for inflammation, mitochondria dynamics, and metabolism. The observed changes were mainly apparent in the liver and typical of activation of type I interferon and NF-κB-dependent signaling, with alterations in mitochondrial dynamics and a shift toward glycolysis. Although the importance of these alterations for the pathophysiology of cancer-related fatigue remains to be explored, the present findings indicate that fatigue brought on by cancer therapy in tumor-bearing mice is more physical than motivational.

Chronic restraint stress impairs voluntary wheel running but has no effect on food-motivated behavior in mice.

Chronic restraint stress is known to cause significant alterations of mitochondrial biology. However, its effects on effort-based behavior and the sensitivity of these effects to treatments that restore mitochondrial function have not been assessed. Based on the hypothesis that the behavioral consequences of this stressor should be more severe for an energy demanding activity than for an energy procuring activity, we compared the effects of chronic restraint stress on the performance of male mice trained to use a running wheel or to nose poke for a food reward in an operant conditioning cage. In accordance with our hypothesis, we observed that exposure of mice to 2-hour daily restraint sessions for 14 to 16 days during the light phase of the cycle reliably decreased voluntary wheel running but had no effect on working for food in a fixed ratio 10 schedule of food reinforcement or in a progressive ratio schedule of food reinforcement. This dissociation between the two types of behavioral activities could reflect an adaptive response to the constraint imposed by chronic restraint stress on mitochondria function and its negative consequences on energy metabolism. To determine whether it is the case, we administered mesenchymal stem cells intranasally to chronically restrained mice to repair the putative mitochondrial dysfunction induced by chronic restraint stress. This intervention had no effect on wheel running deficits. Assessment of mitochondrial gene expression in the brain of mice submitted to chronic restraint stress revealed an increase in the expression of genes involved in mitochondrial biology that showed habituation with repetition of daily sessions of restraint stress. These original findings can be interpreted to indicate that chronic restraint stress induces behavioral and mitochondrial adjustments that contribute to metabolic adaptation to this stressor and maintain metabolic flexibility.

Identification of efflux pump inhibitors for Pseudomonas aeruginosa MexAB-OprM via ligand-based pharmacophores, 2D-QSAR, molecular docking, and molecular dynamics approaches.

Efflux pumps have been reported as one of the significant mechanisms by which bacteria evade the effects of multiple antibiotics. The tripartite efflux pump MexAB-OprM in Pseudomonas aeruginosa is one of the most significant multidrug efflux systems due to its broad resistance to antibiotics such as chloramphenicol, fluoroquinolones, lipophilic ß-lactam antibiotics, nalidixic acid, novobiocin, rifampicin, and tetracycline. A promising strategy to overcome this resistance mechanism is to combine antibiotics with efflux pump inhibitors (EPIs), which can increase their intracellular concentration to enhance their biological activities. Based on 143 EPIs with chemically diverse skeletons, the 3D pharmacophore and 2D-QSAR modelings were developed and used for the virtual screening on 9.2 million compounds including ZINC15, DrugBank, and Traditional Chinese Medicine databases to identify new EPIs. The molecular docking was also performed to evaluate the binding affinity of potential EPIs to the distal-binding pocket of MexB and resulted in 611 potential EPIs. The structure-activity relationship analyses suggested that nitrogen heterocyclic compounds, piperazine and pyridine scaffolds, and amide derivatives are the most favorable chemically features for MexAB inhibitory activities. The results from molecular dynamics analysis in 100 ns indicated that ZINC009296881 and ZINC009200074 were the most potential MexB inhibitors with strong binding affinity to the distal pocket and MM/GBSA ∆Gbind values of - 38.97 and - 30.19 kcal mol-1, respectively. The predicted pharmacokinetic properties and toxicity of these compounds indicated their potential oral drugs. Multistep virtual screening of EPIs for MexAB-OprM, efflux pump multidrug resistant of P. aeruginosa.

Comparative Analysis of Audio Processing Techniques on Doppler Radar Signature of Human Walking Motion Using CNN Models.

Artificial intelligence (AI) radar technology offers several advantages over other technologies, including low cost, privacy assurance, high accuracy, and environmental resilience. One challenge faced by AI radar technology is the high cost of equipment and the lack of radar datasets for deep-learning model training. Moreover, conventional radar signal processing methods have the obstacles of poor resolution or complex computation. Therefore, this paper discusses an innovative approach in the integration of radar technology and machine learning for effective surveillance systems that can surpass the aforementioned limitations. This approach is detailed into three steps: signal acquisition, signal processing, and feature-based classification. A hardware prototype of the signal acquisition circuitry was designed for a Continuous Wave (CW) K-24 GHz frequency band radar sensor. The collected radar motion data was categorized into non-human motion, human walking, and human walking without arm swing. Three signal processing techniques, namely short-time Fourier transform (STFT), mel spectrogram, and mel frequency cepstral coefficients (MFCCs), were employed. The latter two are typically used for audio processing, but in this study, they were proposed to obtain micro-Doppler spectrograms for all motion data. The obtained micro-Doppler spectrograms were then fed to a simplified 2D convolutional neural networks (CNNs) architecture for feature extraction and classification. Additionally, artificial neural networks (ANNs) and 1D CNN models were implemented for comparative analysis on various aspects. The experimental results demonstrated that the 2D CNN model trained on the MFCC feature outperformed the other two methods. The accuracy rate of the object classification models trained on micro-Doppler features was 97.93%, indicating the effectiveness of the proposed approach.

Current Mechanistic Understandings of Lymphedema and Lipedema: Tales of Fluid, Fat, and Fibrosis.

Lymphedema and lipedema are complex diseases. While the external presentation of swollen legs in lower-extremity lymphedema and lipedema appear similar, current mechanistic understandings of these diseases indicate unique aspects of their underlying pathophysiology. They share certain clinical features, such as fluid (edema), fat (adipose expansion), and fibrosis (extracellular matrix remodeling). Yet, these diverge on their time course and known molecular regulators of pathophysiology and genetics. This divergence likely indicates a unique route leading to interstitial fluid accumulation and subsequent inflammation in lymphedema versus lipedema. Identifying disease mechanisms that are causal and which are merely indicative of the condition is far more explored in lymphedema than in lipedema. In primary lymphedema, discoveries of genetic mutations link molecular markers to mechanisms of lymphatic disease. Much work remains in this area towards better risk assessment of secondary lymphedema and the hopeful discovery of validated genetic diagnostics for lipedema. The purpose of this review is to expose the distinct and shared (i) clinical criteria and symptomatology, (ii) molecular regulators and pathophysiology, and (iii) genetic markers of lymphedema and lipedema to help inform future research in this field.

Neuroimmune mechanisms of cognitive impairment in a mouse model of Gulf War illness.

Gulf War Illness (GWI) is a chronic, multi-symptom disorder affecting approximately 30 percent of the nearly 700,000 Veterans of the 1991 Persian Gulf War. GWI-related chemical (GWIC) exposure promotes immune activation that correlates with cognitive impairment and other symptoms of GWI. However, the molecular mechanisms and signaling pathways linking GWIC to inflammation and neurological symptoms remain unclear. Here we show that acute exposure of murine macrophages to GWIC potentiates innate immune signaling and inflammatory cytokine production. Using an established mouse model of GWI, we report that neurobehavioral changes and neuroinflammation are attenuated in mice lacking the cyclic GMP-AMP synthase (cGAS)-Stimulator of Interferon Genes (STING) and NOD-, LRR- or pyrin domain-containing protein 3 (NLRP3) innate immune pathways. In addition, we report sex differences in response to GWIC, with female mice showing more pronounced cognitive impairment and hippocampal astrocyte hypertrophy. In contrast, male mice display a GWIC-dependent upregulation of proinflammatory cytokines in the plasma that is not present in female mice. Our results indicate that STING and NLRP3 are key mediators of the cognitive impairment and inflammation observed in GWI and provide important new information on sex differences in this model.

Outbreaks, occurrence, and control of norovirus and hepatitis a virus contamination in berries: A review.

Foodborne enteric viruses, in particular HuNoV and HAV, are the most common cause of the berry-linked viral diseases, and outbreaks around the world, and have become an important concern for health authorities. Despite the increased importance of berry fruits as a vehicle for foodborne viruses, there is limited information concerning the fate of foodborne viruses in the berry supply chain from farm to consumer. A comprehensive understanding of berry-associated viral outbreaks - with a focus on contamination sources, persistence, survival, and the effects of current postharvest and processing interventions and practices - is essential for the development of effective preventative strategies to reduce risk of illness. The purpose of this paper is twofold; (i) to critically review the published literature on the current state of knowledge regarding berry-associated foodborne viral outbreaks and the efficiency of berry processing practices and (ii) to identify and prioritize research gaps regarding practical and effective mechanism to reduce viral contamination of berries. The review found that fecally infected food handlers were the predominant source of preharvest and postharvest pathogenic viral contamination. Current industrial practices applied to fresh and frozen berries demonstrated limited efficacy for reducing the viral load. While maintaining best practice personal and environmental hygiene is a key intervention, the optimization of processing parameters (i.e., freezing, frozen storage, and washing) and/or development of alternative processing technologies to induce sufficient viral inactivation in berries along with retaining sensory and nutritional quality, is also an important direction for further research.

Prediction model of human ABCC2/MRP2 efflux pump inhibitors: a QSAR study.

The overexpression of ABCC2/MRP2, an ATP-binding cassette transporter, contributes to multidrug resistance in cancer cells. In this study, a quantitative structure-activity relationship (QSAR) analysis on ABCC2 inhibitors has been carried out, aiming to establish a computational prediction model for ABCC2 modulators. Seven classification models and two regression models were built by SONNIA 4.2, and two other regression models were built by MOE 2008.10 based on a data set comprising 372 compounds collected from 16 relevant publications. The CPG-C iABCC2 model for classifying ABCC2 inhibitors has total accuracy of 0.88 and Matthews correlation coefficient MCC = 0.75. The CPG-C iEG model for classifying ABCC2 inhibitors (substrate EG: ß-estradiol 17-ß-D-glucuronide) has total accuracy of 0.91 and MCC = 0.82. The regression model PLS EG-IC50 for predicting ABCC2 inhibitors (substrate EG) gave root-mean-square error RMSE = 0.26, Q2 = 0.73 and [Formula: see text]. The regression model PLS CDCF-IC50 for predicting ABCC2 inhibitors [substrate CDCF: 5(6)-carboxy-2',7'-dichlorofluorescein] gave RMSE = 0.31, Q2 = 0.74 and [Formula: see text]. Four 2D-QSAR models were applied to 1661 compounds, with results indicating 369 compounds having the ability to reverse the efflux of both EG and CDCF by ABCC2, 152 among them having IC50 < 100 µM.

Noninvasive and real-time in vivo characterization of Inflammation skin. A feasibility of animal study.

BACKGROUND: Inflammatory skin diseases were the most common problem in dermatology. This study aimed to develop a circuit by using a simple method for noninvasive, objective, and real-time skin inflammation screening. MATERIALS AND METHODS: Sprague-Dawley rats were used in this study. The rats were chemically induced to suffer from skin inflammation at the back of their left-hand side while the right-hand side of their back remained untreated serving as a control. Impedance (Z) spectrum of the rat's skin was recorded. RESULTS: Two characteristic frequencies (4.5 and 48.3 kHz) were found. At the two frequencies, the impedance of inflammatory skin tissue (ZIST ) was found to be significantly (P < .05) smaller than that of normal healthy skin tissue (ZNHST ). Moreover, the ratio of the impedance measured at 4.5 kHz (Zf = 4 .5 kHz ) to the impedance measured at 48.3 kHz (Zf = 48.3 kHz ), that is, Zf = 4.5 kHz /Zf = 48.3 kHz , was capable of skin inflammation screening. It was observed that the inflammatory skin tissue (IST) had the smaller value of Zf = 4 .5 kHz /Zf = 48.3 kHz (value < 8.5) and normal healthy skin tissue (NHST) had the higher value of Zf = 4 .5 kHz /Zf = 48.3 kHz (value ≈ 10) which almost remained constant. CONCLUSION: A circuit was developed which was used for measuring the skin impedance accurately at the two characteristic frequencies for skin inflammation screening.

Assessment of microbial risk during Australian industrial practices for Escherichia coli O157:H7 in fresh cut-cos lettuce: A stochastic quantitative approach.

Escherichia coli O157:H7 risk associated with the consumption of fresh cut-cos lettuce during Australian industrial practices was assessed. A probabilistic risk assessment model was developed and implemented in the @Risk software by using the Monte Carlo simulation technique with 1,000,000 iterations. Australian preharvest practices yielded predicted annual mean E. coli O157:H7 levels from 0.2 to -3.4 log CFU/g and prevalence values ranged from 2 to 6.4%. While exclusion of solar radiation from the baseline model yielded a significant increase in concentration of E. coli O157:H7 (-5.2 -log fold), drip irrigation usage, exclusion of manure amended soil and rainfall reduced E. coli O157:H7 levels by 7.4, 6.5, and 4.3-log fold, respectively. The microbial quality of irrigation water and irrigation type both had a significant effect on E. coli O157:H7 concentrations at harvest (p < 0.05). The probability of illness due to consumption of E. coli O157:H7 contaminated fresh cut-cos lettuce when water washing interventions were introduced into the processing module, was reduced by 1.4-2.7-log fold (p < 0.05). This study provides a robust basis for assessment of risk associated with E. coli O157:H7 contamination on fresh cut-cos lettuce for industrial practices and will assist the leafy green industry and food safety authorities in Australia to identify potential risk management strategies.

Structure-Based Discovery of ABCG2 Inhibitors: A Homology Protein-Based Pharmacophore Modeling and Molecular Docking Approach.

ABCG2 is an ABC membrane protein reverse transport pump, which removes toxic substances such as medicines out of cells. As a result, drug bioavailability is an unexpected change and negatively influences the ADMET (absorption, distribution, metabolism, excretion, and toxicity), leading to multi-drug resistance (MDR). Currently, in spite of promising studies, screening for ABCG2 inhibitors showed modest results. The aim of this study was to search for small molecules that could inhibit the ABCG2 pump. We first used the WISS MODEL automatic server to build up ABCG2 homology protein from 655 amino acids. Pharmacophore models, which were con-structed based on strong ABCG2 inhibitors (IC50 < 1 µM), consist of two hydrophobic (Hyd) groups, two hydrogen bonding acceptors (Acc2), and an aromatic or conjugated ring (Aro|PiR). Using molecular docking method, 714 substances from the DrugBank and 837 substances from the TCM with potential to inhibit the ABCG2 were obtained. These chemicals maybe favor synthesized or extracted and bioactivity testing.

Demonstration of a laser powder bed fusion combinatorial sample for high-throughput microstructure and indentation characterization.

High-throughput experiments that use combinatorial samples with rapid measurements can be used to provide process-structure-property information at reduced time, cost, and effort. Developing these tools and methods is essential in additive manufacturing where new process-structure-property information is required on a frequent basis as advances are made in feedstock materials, additive machines, and post-processing. Here we demonstrate the design and use of combinatorial samples produced on a commercial laser powder bed fusion system to study 60 distinct process conditions of nickel superalloy 625: five laser powers and four laser scan speeds in three different conditions. Combinatorial samples were characterized using optical and electron microscopy, x-ray diffraction, and indentation to estimate the porosity, grain size, crystallographic texture, secondary phase precipitation, and hardness. Indentation and porosity results were compared against a regular sample. The smaller-sized regions (3 mm × 4 mm) in the combinatorial sample have a lower hardness compared to a larger regular sample (20 mm × 20 mm) with similar porosity (< 0.03 %). Despite this difference, meaningful trends were identified with the combinatorial sample for grain size, crystallographic texture, and porosity versus laser power and scan speed as well as trends with hardness versus stress-relief condition.

Gender-Related Differences of Tachykinin NK2 Receptor Expression and Activity in Human Colonic Smooth Muscle.

The tachykinin NK2 receptor plays a key role in gastrointestinal motor function. Enteric neurons release neurokinin A (NKA), which activates NK2 receptors on gastrointestinal smooth muscle, leading to contraction and increased motility. In patients with diarrhea-predominant irritable bowel syndrome, the NK2 receptor antagonist ibodutant had a greater therapeutic effect in females than males. The present study aimed to determine whether gender influences the expression and activity of NK2 receptors in human colonic smooth muscle. In vitro functional studies were performed to examine the contractile responses of colonic muscle strips to NKA and the selective NK2 receptor agonist [Lys5,MeLeu9,Nle10]NKA(4-10). Contractions were also measured in the presence of ibodutant to determine its antagonistic potency. The signal transduction pathways coupled to NK2 receptor activation were investigated using second messenger inhibitors. Western blot and fluorescent immunohistochemistry were conducted to determine the protein expression and localization of NK2 receptors. NK2 receptor-mediated contractility was greater in females compared with males. When against NKA, ibodutant was more potent in females. NK2 receptor expression increased with age in females, but not in males. Phospholipase C-mediated signaling was less prominent in females compared with males, whereas Ca2+ sensitization via Rho kinase and protein kinase C appeared to be the dominant pathway in both genders. The distribution of NK2 receptors in the human colon did not differ between the genders. Overall, gender differences exist in the expression and activity of NK2 receptors in colonic smooth muscle. These gender distinctions should be considered in the therapeutic development of NK2 receptor agents. SIGNIFICANCE STATEMENT: The tachykinin NK2 receptor has been identified as a therapeutic target for the treatment of bowel and bladder dysfunctions. The present study has revealed gender-related variations in NK2 receptor activity, signaling transduction pathways, antagonist potency, and changes in expression with age. These factors may underlie the gender differences in the treatment of diarrhea-predominant irritable bowel syndrome with NK2 receptor antagonists. Our findings highlight that gender differences should be considered in the therapeutic development of NK2 receptor agents.

Thermoresponsive Hydrogel Induced by Dual Supramolecular Assemblies and Its Controlled Release Property for Enhanced Anticancer Drug Delivery.

Supramolecular hydrogels based on inclusion complexation between cyclodextrins (CDs) and polymers have attracted much interest because of their potential for biomedical applications. It is also attractive to incorporate stimuli-responsive properties into the system to create "smart" hydrogels. Herein, a poly(N-isopropylacrylamide) (PNIPAAm) star polymer with a ß-CD core and an adamantyl-terminated poly(ethylene glycol) (Ad-PEG) polymer were synthesized. They self-assembled into a thermoresponsive pseudo-block copolymer through host-guest complexation and formed supramolecular micelles with the change in environment temperature. Subsequently, an injectable polypseudorotaxane-based supramolecular hydrogel was formed between α-CD and the PEG chains of the pseudo-block copolymer. The hydrogel had a unique network structure involving two types of supramolecular self-assemblies between cyclodextrins and polymers, that is, the host-guest complexation between ß-CD units and adamantyl groups and the polypseudorotaxane formation between α-CD and PEG chains. We hypothesize that the dual supramolecular hydrogel formed at room temperature may be enhanced by increasing the temperature over the lower critical solution temperature of PNIPAAm because of the hydrophobic interactions of PNIPAAm segments. Furthermore, if the hydrogel is applied for sustained delivery of hydrophobic drugs, the copolymer dissolved from the hydrogel could micellize and continue to serve as micellar drug carriers with the drug encapsulated in the hydrophobic core. Rheological tests revealed that the hydrophobic interactions of the PNIPAAm segments could significantly enhance the strength of the hydrogel when the temperature increased from 25 to 37 °C. As compared to hydrogels formed by α-CD and PEG alone, the sustained release property of this thermoresponsive hydrogel for an anticancer drug, doxorubicin (DOX), improved at 37 °C. The hydrogel dissolved slowly and released the pseudo-block copolymer in the form of micelles that continued to serve as drug carriers with DOX encapsulated in the hydrophobic core, achieving a better cellular uptake and anticancer effect than free DOX controls, even in multidrug-resistant cancer cells. According to these findings, the dual supramolecular hydrogel developed in this work with remarkable thermoresponsive properties might have potential for sustained anticancer drug delivery with enhanced therapeutic effect in multidrug-resistant cancer cells.

Effect of soil type and temperature on survival of Salmonella enterica in poultry manure-amended soils.

The effects of soil type and temperature on the survival of a cocktail of five Salmonella enterica serotypes (Enteritidis, Infantis, Montevideo, Typhimurium and Zanzibar) in manure-amended soils under controlled laboratory conditions was assessed. Containers of clay loam or sandy soil, unaltered or amended with 2% (w/w) poultry manure, were inoculated with S. enterica (~5 log10 CFU per gram) and held at 5, 21 or 37°C for 6 weeks. Statistical analysis of the persistence of S. enterica identified a significant three-way interaction between soil type, manure amendment and temperature. Clay loam soils and lower temperatures tended to support S. enterica persistence over 6 weeks with only 1- and 2-log reductions respectively. In contrast, sand and higher temperatures resulted in a 4-log and either 3- to 4-log reductions respectively. Manure amendment had an overarching effect of reducing die-off of S. enterica in comparison with unamended soils. This study highlights that a large component of variation of the rate of S. enterica reduction in soils may be attributed to combinations of environmental factors, in particular, soil type and temperature. It further underscores the importance of risk management strategies and industry guidelines based on local data and that reflect the diversity of prevailing horticultural production environments. SIGNIFICANCE AND IMPACT OF THE STUDY: The persistence of Salmonella enterica in soil environments was shown to be significantly influenced by a range of individual and interacting environmental effects, including temperature, soil type and amendment addition. This indicates that current horticultural food safety management systems which employ a uniform prescribed exclusion period between application of manure and time of harvest may be unfit for purpose under certain conditions by either underestimating or overestimating pathogen die-off. These findings support exclusion periods that account for a range of environmental factors including temperature, soil type and growing region that may be more appropriate to manage microbiological risks associated with soil which has been amended with manure.

Subtotal colectomy and ileorectal anastomosis for slow transit constipation: clinical follow-up at median of 15 years.

BACKGROUND: Slow transit constipation is characterised by prolonged colonic transit and reliance on laxatives. The pathophysiology is poorly understood and in its most severe form, total colectomy with ileorectal anastomosis is the final treatment option. We present a follow-up study of the long-term function in patients who had surgery for laxative-resistant slow transit constipation. METHODS: A postal survey was sent to assess bowel frequency, abdominal pain, St Mark's continence score, satisfaction with procedure, likelihood to choose the procedure again, and long-term rates of small bowel obstruction and ileostomy. Longitudinal data from a subgroup studied 23 years previously are reported. RESULTS: Forty-two patients (male = 2) were available for follow-up out of an initial cohort of 102. Mean time since surgery was 15.9 years (range 1.7-29.7) years. Fifty percent had < 4 bowel motions per day, most commonly Bristol stool 6, mean St Mark's score 7.45. Twenty-one percent had severe incontinence. Satisfaction and likelihood to choose surgery were high (median 10/10). There was a high rate of small bowel obstruction, suggesting pan-intestinal dysmotility in some cases. Conversion to ileostomy occurred in 8 patients. In the longitudinal follow-up in 15 subjects, continence deteriorated (p < 0.01), stool consistency softened (p < 0.01), and stool frequency fell (p < 0.01). CONCLUSIONS: Satisfactory stool frequency was achieved in the long term, and although 21% had incontinence scores > 12, patient satisfaction was high. This is the longest reported follow-up of colectomy for slow transit constipation, with longitudinal outcomes reported. There was considerable attrition of patients, so larger, longitudinal studies are required to better ascertain the functional outcomes of these patients.

Process evaluation of a behaviour change approach to improving clinical practice for detecting hereditary cancer.

BACKGROUND: This retrospective process evaluation reports on the application of a 1-year implementation program to increase identification and management of patients at high risk of a hereditary cancer syndrome. The project used the Theoretical Domains Framework Implementation (TDFI) approach, a promising implementation methodology, used successfully in the United Kingdom to address patient safety issues. This Australian project run at two large public hospitals aimed to increase referrals of patients flagged as being at risk of Lynch syndrome on the basis of a screening test to genetic services. At the end of the project, the pathologists' processes had changed, but the referral rate remained inconsistent and low. METHODS: Semi-structured interviews explored participants' perceptions of the TDFI approach and Health services researchers wrote structured reflections. Interview transcripts and reflections were coded initially against implementation outcomes for the various TDFI approach activities: acceptability, appropriateness, feasibility, value for time cost, and adoption. On a second pass, themes were coded around challenges to the approach. RESULTS: Interviews were held with nine key project participants including pathologists, oncologists, surgeons, genetic counsellors and an administrative officer. Two health services researchers wrote structured reflections. The first of two major themes was 'Theory-related challenges', with subthemes of accessibility of theory underpinning the TDFI, commitment to that theory-based approach, and the problem of complexity. The second theme was 'Practical challenges' with subthemes of stakeholder management, navigating the system, and perceptions of the problem. Health services researchers reflected on the benefits of bridging professional divides and facilitating collective learning and problem solving, but noted frustrations around clinicians' time constraints that led to sparse interactions with the team, and lack of authority to effect change themselves. CONCLUSIONS: Mixed success of adoption as an outcome was attributed to the complexity and highly nuanced nature of the setting. This made identifying the target behaviour, a key step in the TDFI approach, challenging. Introduced changes in the screening process led to new, unexpected issues yet to be addressed. Strategies to address challenges are presented, including using an internal facilitator with a focus on applying a theory-based implementation approach.

Quantifying Vascular Distribution and Adhesion of Nanoparticles with Protein Corona in Microflow.

The protein corona has emerged as an important determinant of biological response in nanoparticle (NP) drug delivery. However, there is presently no reported study on how the protein corona affects the behavior of NPs in microflow and its subsequent interactions with the vascular endothelium, which could affect their delivery to the target tumor site regardless of its targeting mechanism. Furthermore, a consensus on the role of physical and surface characteristics of NPs in affecting the margination of NPs is lacking due to different methods of quantifying margination. In this study, we examine how the particle adhesion (PA) method and particle distribution (PD) method quantify the margination of 20, 40, 100, and 200 nm polystyrene NPs (pNPs) differently in fibronectin or pluronic F-127-coated microfluidic straight channels. We found that PA reduced with increasing pNP size, whereas the PD was similar across all pNP sizes regardless of channel coating. We then formed a protein corona on all pNPs (pNPs-PC) and found that the protein corona increased the adhesion of 40-200 nm pNPs in fibronectin-coated channels, with no size dependence between them except for 40 nm, which had significantly higher particle adhesion. The PA method was also dependent on channel coating, whereas the PD method was independent of channel coating. These results suggested that the PA method was more amenable to surface interactions between the pNPs and the channel wall while providing a measure of the amount of NPs that interacted with the channel walls, whereas the PD method provided a representation of their distribution across the channel due to margination. The two methods complement each other to elucidate a more holistic understanding of how different factors might affect a NP's margination in future studies.

Relationship between transit time and mechanical properties of a cell through a stenosed microchannel.

The changes in the mechanical properties of a cell are not only the cause of some diseases, but can also be a biomarker for some disease states. In recent times, microfluidic devices with built-in constrictions have been widely used to measure these changes. The transit time in such devices, defined as the time that a cell takes to pass through a constriction, has been found to be a crucial factor associated with the cell mechanical properties. Here, we use smoothed dissipative particle dynamics (SDPD), a particle-based numerical method, to explore the relationship between the transit time and mechanical properties of a cell. Three expressions of the transit time are developed from our simulation data, with respect to the stenosed size of constrictions, the shear modulus and bending modulus of cells, respectively. We show that a convergent constriction (the inlet is wider than the outlet), and a sharp-corner constriction (the constriction outlet is narrow) are better in identifying the differences in the transit time of cells. Moreover, the transit time increases and gradually approaches a constant as the shear modulus of cells increases, but increases first and then decreases as the bending modulus increases. These results suggest that the mechanical properties of cells can indeed be measured by analyzing their transit time, based on the recommended microfluidic device.

Using behaviour change and implementation science to address low referral rates in oncology.

BACKGROUND: Patients undergoing surgery for bowel cancer now have a routine screening test to assess their genetic predisposition to this and other cancers (Lynch syndrome). A result indicating a high risk should trigger referral to a genetic clinic for diagnostic testing, information, and management. Appropriate management of Lynch syndrome lowers morbidity and mortality from cancer for patients and their family, but referral rates are low. The aim of this project was to increase referral rates for patients at high risk of Lynch syndrome at two Australian hospitals, using the Theoretical Domains Framework (TDF) Implementation approach. METHODS: Multidisciplinary teams at each hospital mapped the referral process and discussed barriers to referral. A 12-month retrospective audit measured baseline referral rates. The validated Influences on Patient Safety Behaviours Questionnaire was administered to evaluate barriers using the TDF. Results were discussed in focus groups and interviews, and interventions co-designed, guided by theory. Continuous monitoring audits assessed change in referral rates. RESULTS: Teams (n = 8, 11) at each hospital mapped referral processes. Baseline referral rates were 80% (4/5) from 71 screened patients and 8% (1/14) from 113 patients respectively. The questionnaire response rate was 51% (36/71). Most significant barrier domains were: 'environmental context;' 'memory and decision making;' 'skills;' and 'beliefs about capabilities.' Focus groups and interviews with 19 healthcare professionals confirmed these domains as significant. Fifteen interventions were proposed considering both emerging and theory-based results. Interventions included: clarification of pathology reports, education, introduction of e-referrals, and inclusion of genetic status in documentation. Audits continued to December 2016 showing a change in pathology processes which increased the accuracy of screening. The referral rate remained low: 46% at Hospital A and 9% Hospital B. Results suggest patients who have their referral deferred for some reason are not referred later. CONCLUSION: Lynch syndrome is typical of low incidence problems likely to overwhelm the system as genomic testing becomes mainstream. It is crucial for health researchers to test methods and define generalizable solutions to address this problem. Whilst our approach did not improve referrals, we have deepened our understanding of barriers to referral and approaches to low frequency conditions.