Nano-liter perfusion microfluidic device made entirely by two-photon polymerization for dynamic cell culture with easy cell recovery.

Polydimethylsiloxane (PDMS) has been the material of choice for microfluidic applications in cell biology for many years, with recent advances encompassing nano-scaffolds and surface modifications to enhance cell-surface interactions at nano-scale. However, PDMS has not previously been amenable to applications which require complex geometries in three dimensions for cell culture device fabrication in the absence of additional components. Further, PDMS microfluidic devices have limited capacity for cell retrieval following culture without severely compromising cell health. This study presents a designed and entirely 3D-printed microfluidic chip (8.8 mm × 8.2 mm × 3.6 mm) using two-photon polymerization (2PP). The 'nest' chip is composed of ten channels that deliver sub-microliter volume flowrates (to ~ 600 nL/min per channel) to 10 individual retrievable cell sample 'cradles' that interlock with the nest to create the microfluidic device. Computational fluid dynamics modelling predicted medium flow in the device, which was accurately validated by real-time microbead tracking. Functional capability of the device was assessed, and demonstrated the capability to deliver culture medium, dyes, and biological molecules to support cell growth, staining and cell phenotype changes, respectively. Therefore, 2PP 3D-printing provides the precision needed for nanoliter fluidic devices constructed from multiple interlocking parts for cell culture application.

Epichloë seed transmission efficiency is influenced by plant defense response mechanisms.

Asexual Epichloë are endophytic fungi that form mutualistic symbioses with cool-season grasses, conferring to their hosts protection against biotic and abiotic stresses. Symbioses are maintained between grass generations as hyphae are vertically transmitted from parent to progeny plants through seed. However, endophyte transmission to the seed is an imperfect process where not all seeds become infected. The mechanisms underpinning the varying efficiencies of seed transmission are poorly understood. Host gene expression in response to Epichloë sp. LpTG-3 strain AR37 was examined within inflorescence primordia and ovaries of high and low endophyte transmission genotypes within a single population of perennial ryegrass. A genome-wide association study was conducted to identify population-level single nucleotide polymorphisms (SNPs) and associated genes correlated with vertical transmission efficiency. For low transmitters of AR37, upregulation of perennial ryegrass receptor-like kinases and resistance genes, typically associated with phytopathogen detection, comprised the largest group of differentially expressed genes (DEGs) in both inflorescence primordia and ovaries. DEGs involved in signaling and plant defense responses, such as cell wall modification, secondary metabolism, and reactive oxygen activities were also abundant. Transmission-associated SNPs were associated with genes for which gene ontology analysis identified "response to fungus" as the most significantly enriched term. Moreover, endophyte biomass as measured by quantitative PCR of Epichloë non-ribosomal peptide synthetase genes, was significantly lower in reproductive tissues of low-transmission hosts compared to high-transmission hosts. Endophyte seed-transmission efficiency appears to be influenced primarily by plant defense responses which reduce endophyte colonization of host reproductive tissues.

Outlier analyses and genome-wide association study identify glgC and ERD6-like 4 as candidate genes for foliar water-soluble carbohydrate accumulation in Trifolium repens.

Increasing water-soluble carbohydrate (WSC) content in white clover is important for improving nutritional quality and reducing environmental impacts from pastoral agriculture. Elucidation of genes responsible for foliar WSC variation would enhance genetic improvement by enabling molecular breeding approaches. The aim of the present study was to identify single nucleotide polymorphisms (SNPs) associated with variation in foliar WSC in white clover. A set of 935 white clover individuals, randomly sampled from five breeding pools selectively bred for divergent (low or high) WSC content, were assessed with 14,743 genotyping-by-sequencing SNPs, using three outlier detection methods: PCAdapt, BayeScan and KGD-FST. These analyses identified 33 SNPs as discriminating between high and low WSC populations and putatively under selection. One SNP was located in the intron of ERD6-like 4, a gene coding for a sugar transporter located on the vacuole membrane. A genome-wide association study using a subset of 605 white clover individuals and 5,757 SNPs, identified a further 12 SNPs, one of which was associated with a starch biosynthesis gene, glucose-1-phosphate adenylyltransferase, glgC. Our results provide insight into genomic regions underlying WSC accumulation in white clover, identify candidate genomic regions for further functional validation studies, and reveal valuable information for marker-assisted or genomic selection in white clover.

Deterministic and stochastic modelling of impacts from genomic selection and phenomics on genetic gain for perennial ryegrass dry matter yield.

Increasing the efficiency of current forage breeding programs through adoption of new technologies, such as genomic selection (GS) and phenomics (Ph), is challenging without proof of concept demonstrating cost effective genetic gain (∆G). This paper uses decision support software DeltaGen (tactical tool) and QU-GENE (strategic tool), to model and assess relative efficiency of five breeding methods. The effect on ∆G and cost ($) of integrating GS and Ph into an among half-sib (HS) family phenotypic selection breeding strategy was investigated. Deterministic and stochastic modelling were conducted using mock data sets of 200 and 1000 perennial ryegrass HS families using year-by-season-by-location dry matter (DM) yield data and in silico generated data, respectively. Results demonstrated short (deterministic)- and long-term (stochastic) impacts of breeding strategy and integration of key technologies, GS and Ph, on ∆G. These technologies offer substantial improvements in the rate of ∆G, and in some cases improved cost-efficiency. Applying 1% within HS family GS, predicted a 6.35 and 8.10% ∆G per cycle for DM yield from the 200 HS and 1000 HS, respectively. The application of GS in both among and within HS selection provided a significant boost to total annual ∆G, even at low GS accuracy rA of 0.12. Despite some reduction in ∆G, using Ph to assess seasonal DM yield clearly demonstrated its impact by reducing cost per percentage ∆G relative to standard DM cuts. Open-source software tools, DeltaGen and QuLinePlus/QU-GENE, offer ways to model the impact of breeding methodology and technology integration under a range of breeding scenarios.

Mechanistic insights into the formation of polyion complex aggregates from cationic thermoresponsive diblock copolymers.

HYPOTHESIS: The formation of polyion complexes (PICs) comprising thermoresponsive polymers is intended to result in the formation of aggregates that undergo significant structural changes with temperature. Moreover the observed modifications might be critically affected by polymer structure and PICs composition. EXPERIMENTS: Different block copolymers based on cationic poly(3-acrylamidopropyltrimethylammonium chloride) and thermoresponsive poly(N-isopropylacrylamide) were synthesized by aqueous RAFT/MADIX polymerization at room temperature. Addition of poly(acrylic acid) in a controlled fashion led to the formation of PICs aggregates. The structural changes induced by temperature were characterized by differential scanning calorimetry, Nuclear Magnetic Resonance spectroscopy and scattering methods. FINDINGS: Thermoresponsive PICs undergo significant structural changes when increasing temperature above the cloud point of the thermoresponsive block. The reversibility of these phenomena depends strongly on the structural parameters of the block copolymers and on PICs composition.

Multi-Trait Genomic Prediction Improves Predictive Ability for Dry Matter Yield and Water-Soluble Carbohydrates in Perennial Ryegrass.

In perennial ryegrass (Lolium perenne L), annual and seasonal dry matter yield (DMY) and nutritive quality of herbage are high-priority traits targeted for improvement through selective breeding. Genomic prediction (GP) has proven to be a valuable tool for improving complex traits and may be further enhanced through the use of multi-trait (MT) prediction models. In this study, we evaluated the relative performance of MT prediction models to improve predictive ability for DMY and key nutritive quality traits, using two different training populations (TP1, n = 463 and TP2, n = 517) phenotyped at multiple locations. MT models outperformed single-trait (ST) models by 24% to 59% for DMY and 67% to 105% for nutritive quality traits, such as low, high, and total WSC, when a correlated secondary trait was included in both the training and test set (MT-CV2) or in the test set alone (MT-CV3) (trait-assisted genomic selection). However, when a secondary trait was included in training set and not the test set (MT-CV1), the predictive ability was not statistically significant (p > 0.05) compared to the ST model. We evaluated the impact of training set size when using a MT-CV2 model. Using a highly correlated trait (rg = 0.88) as the secondary trait in the MT-CV2 model, there was no loss in predictive ability for DMY even when the training set was reduced to 50% of its original size. In contrast, using a weakly correlated secondary trait (rg = 0.56) in the MT-CV2 model, predictive ability began to decline when the training set size was reduced by only 11% from its original size. Using a ST model, genomic predictive ability in a population unrelated to the training set was poor (rp = -0.06). However, when using an MT-CV2 model, the predictive ability was positive and high (rp = 0.76) for the same population. Our results demonstrate the first assessment of MT models in forage species and illustrate the prospects of using MT genomic selection in forages, and other outcrossing plant species, to accelerate genetic gains for complex agronomical traits, such as DMY and nutritive quality characteristics.

Genomic Predictive Ability for Foliar Nutritive Traits in Perennial Ryegrass.

Forage nutritive value impacts animal nutrition, which underpins livestock productivity, reproduction and health. Genetic improvement for nutritive traits in perennial ryegrass has been limited, as they are typically expensive and time-consuming to measure through conventional methods. Genomic selection is appropriate for such complex and expensive traits, enabling cost-effective prediction of breeding values using genome-wide markers. The aims of the present study were to assess the potential of genomic selection for a range of nutritive traits in a multi-population training set, and to quantify contributions of family, location and family-by-location variance components to trait variation and heritability for nutritive traits. The training set consisted of a total of 517 half-sibling (half-sib) families, from five advanced breeding populations, evaluated in two distinct New Zealand grazing environments. Autumn-harvested samples were analyzed for 18 nutritive traits and maternal parents of the half-sib families were genotyped using genotyping-by-sequencing. Significant (P < 0.05) family variance was detected for all nutritive traits and genomic heritability (h2g ) was moderate to high (0.20 to 0.74). Family-by-location interactions were significant and particularly large for water soluble carbohydrate (WSC), crude fat, phosphorus (P) and crude protein. GBLUP, KGD-GBLUP and BayesCπ genomic prediction models displayed similar predictive ability, estimated by 10-fold cross validation, for all nutritive traits with values ranging from r = 0.16 to 0.45 using phenotypes from across two locations. High predictive ability was observed for the mineral traits sulfur (0.44), sodium (0.45) and magnesium (0.45) and the lowest values were observed for P (0.16), digestibility (0.22) and high molecular weight WSC (0.23). Predictive ability estimates for most nutritive traits were retained when marker number was reduced from one million to as few as 50,000. The moderate to high predictive abilities observed suggests implementation of genomic selection is feasible for most of the nutritive traits examined.

Cultural Immersion in the Education of Healthcare Professionals: A Systematic Review.

PURPOSE: With the ever-changing cultural makeup of society, the ability to deliver culturally appropriate healthcare is essential. One educational method aimed at increasing cultural knowledge and sensitivity in the education of healthcare professionals is cultural immersion. Cultural immersion creates opportunity for transformational learning through direct interaction with culturally diverse populations. The purpose of this systematic review is to examine the qualitative effects of cultural immersion experiences on graduate-level healthcare professional students. METHODS: A search of the CINAHL and ERIC databases was performed utilizing search terms including cultural immersion, cultural sensitivity, educational outcomes, and healthcare professionals limited to publication within the last 10 years. The articles were screened according to title, abstract, and full-text following application of inclusion/exclusion criteria. Themes identified within each article were collected and categorized, using qualitative methodology, into five overarching domains to assess the educational experiences. Studies were scored for quality using the qualitative portion of the McGill Mixed Methods Appraisal Tool - 2011. RESULTS: Nine studies incorporating a total of 94 participants with experiences in 14 different culturally diverse environments revealing 47 individually identified themes were included in the review. Results indicated that all cultural immersion experiences stimulated increased cultural awareness and sensitivity. DISCUSSION: Cultural immersion experiences produced a positive, multi-domain effect on cultural learning in students of the health professionals. Results support a basis for implementation of cultural immersion experiences into the education of healthcare professionals with the goal of increasing cultural sensitivity.

Seed Transmission of Epichloë Endophytes in Lolium perenne Is Heavily Influenced by Host Genetics.

Vertical transmission of symbiotic Epichloë endophytes from host grasses into progeny seed is the primary mechanism by which the next generation of plants is colonized. This process is often imperfect, resulting in endophyte-free seedlings which may have poor ecological fitness if the endophyte confers protective benefits to its host. In this study, we investigated the influence of host genetics and environment on the vertical transmission of Epichloë festucae var. lolii strain AR37 in the temperate forage grass Lolium perenne. The efficiency of AR37 transmission into the seed of over 500 plant genotypes from five genetically diverse breeding populations was determined. In Populations I-III, which had undergone previous selection for high seed infection by AR37, mean transmission was 88, 93, and 92%, respectively. However, in Populations IV and V, which had not undergone previous selection, mean transmission was 69 and 70%, respectively. The transmission values, together with single-nucleotide polymorphism data obtained using genotyping-by-sequencing for each host, was used to develop a genomic prediction model for AR37 seed transmission. The predictive ability of the model was estimated at r = 0.54. While host genotype contributed greatly to differences in AR37 seed transmission, undefined environmental variables also contributed significantly to seed transmission across different years and geographic locations. There was evidence for a small host genotype-by-environment effect; however this was less pronounced than genotype or environment alone. Analysis of endophyte infection levels in parent plants within Populations I and IV revealed a loss of endophyte infection over time in Population IV only. This population also had lower average tiller infection frequencies than Population I, suggesting that AR37 failed to colonize all the daughter tillers and therefore seeds. However, we also observed that infection of seed by AR37 may fail during or after initiation of floral development from plants where all tillers remained endophyte-infected over time. While the effects of environment and host genotype on fungal endophyte transmission have been evaluated previously, this is the first study that quantifies the relative impacts of host genetics and environment on endophyte vertical transmission.

Protocol: a versatile, inexpensive, high-throughput plant genomic DNA extraction method suitable for genotyping-by-sequencing.

BACKGROUND: The recent development of next-generation sequencing DNA marker technologies, such as genotyping-by-sequencing (GBS), generates thousands of informative single nucleotide polymorphism markers in almost any species, regardless of genomic resources. This enables poorly resourced or "orphan" crops/species access to high-density, high-throughput marker platforms which have revolutionised population genetics studies and plant breeding. DNA quality underpins success of GBS methods as the DNA must be amenable to restriction enzyme digestion and sequencing. A barrier to implementing GBS technologies is access to inexpensive, high-throughput extraction methods that yield sequencing-quality genomic DNA (gDNA) from plants. Several high-throughput DNA extraction methods are available, but typically provide low yield or poor quality gDNA, or are costly (US$6-$9/sample) for consumables. RESULTS: We modified a non-organic solvent protocol to extract microgram quantities (1-13 µg) of sequencing-quality high molecular weight gDNA inexpensively in 96-well plates from either fresh, freeze-dried or silica gel-dried plant tissue. The protocol was effective for several easy and difficult-to-extract forage, crop, horticultural and common model species including Trifolium, Medicago, Lolium, Secale, Festuca, Malus, Oryza, and Arabidopsis. The extracted DNA was of high molecular weight and digested readily with restriction enzymes. Contrasting with other extraction protocols we assessed, Illumina-based sequencing of GBS libraries developed from this gDNA had very uniform high quality base-calls to the end of sequence reads. Furthermore, DNA extracted using this method has been sequenced successfully with the PacBio long-read platform. The protocol is scalable, readily automated without requirement for fume hoods, requires approximately three hours to process 192 samples (384-576 samples/day), and is inexpensive at US$0.62/sample for consumables. CONCLUSIONS: This versatile, scalable and simple protocol yields high molecular weight genomic DNA suitable for restriction enzyme digestion and next-generation sequencing applications including GBS and long-read sequencing platforms such as PacBio. The low cost, high-throughput, and extraction of high quality gDNA from a range of fresh and dried source plant material makes this method suitable for many sequencing and genotyping applications including large-scale sample screening underpinning breeding programmes.

Low pyrrolizidine alkaloid levels in perennial ryegrass is associated with the absence of a homospermidine synthase gene.

BACKGROUND: Pyrrolizidine alkaloids (PAs) are a class of secondary metabolites that function as feeding deterrents in a range of different plant species. In perennial ryegrass (Lolium perenne L.) the only PAs that have been identified are the thesinine-rhamnoside group, which displays significant genetic variation. Homospermidine synthase (HSS) has evolved from deoxyhypusine synthase (DHS) and catalyses the first step in the PA pathway, making it a key candidate for the investigation of genes influencing observed PA trait variation. RESULTS: During PCR amplification and sequence analysis of DHS we identified two putative HSS genes in perennial ryegrass. One of the genes (LpHSS1) was absent in some perennial ryegrass plants. Thesinine-rhamnoside levels were measured using liquid chromatography coupled with mass spectrometry in a diverse association mapping population, consisting of 693 plants free of fungal endophytic symbionts. Association tests that accounted for population structure identified a significant association of absence of the LpHSS1 gene with lower levels of thesinine-rhamnoside PAs. HSS-like gene sequences were identified for other grass species of the Poaceae, including tall fescue, wheat, maize and sorghum. CONCLUSION: HSS is situated at the crucial first step in the PA pathway making it an important candidate gene for investigation of involvement in PA phenotypic variation. In this study, PA level in perennial ryegrass was strongly associated with the presence or absence of the LpHSS1 gene. A genetic marker, developed for the presence/absence of LpHSS1, may be used for marker-assisted breeding to either lower or increase PAs in breeding populations of perennial or Italian ryegrass to investigate a potential role in the deterrence of herbivore pests. The presence of HSS-like genes in several other Poaceae species suggests that PA biosynthesis may occur in plant family members beyond perennial ryegrass and tall fescue and identifies a potential route for manipulating PA levels.

Predictive ability of genomic selection models in a multi-population perennial ryegrass training set using genotyping-by-sequencing.

KEY MESSAGE: Genomic prediction models for multi-year dry matter yield, via genotyping-by-sequencing in a composite training set, demonstrate potential for genetic gain improvement through within-half sibling family selection. Perennial ryegrass (Lolium perenne L.) is a key source of nutrition for ruminant livestock in temperate environments worldwide. Higher seasonal and annual yield of herbage dry matter (DMY) is a principal breeding objective but the historical realised rate of genetic gain for DMY is modest. Genomic selection was investigated as a tool to enhance the rate of genetic gain. Genotyping-by-sequencing (GBS) was undertaken in a multi-population (MP) training set of five populations, phenotyped as half-sibling (HS) families in five environments over 2 years for mean herbage accumulation (HA), a measure of DMY potential. GBS using the ApeKI enzyme yielded 1.02 million single-nucleotide polymorphism (SNP) markers from a training set of n = 517. MP-based genomic prediction models for HA were effective in all five populations, cross-validation-predictive ability (PA) ranging from 0.07 to 0.43, by trait and target population, and 0.40-0.52 for days-to-heading. Best linear unbiased predictor (BLUP)-based prediction methods, including GBLUP with either a standard or a recently developed (KGD) relatedness estimation, were marginally superior or equal to ridge regression and random forest computational approaches. PA was principally an outcome of SNP modelling genetic relationships between training and validation sets, which may limit application for long-term genomic selection, due to PA decay. However, simulation using data from the training experiment indicated a twofold increase in genetic gain for HA, when applying a prediction model with moderate PA in a single selection cycle, by combining among-HS family selection, based on phenotype, with within-HS family selection using genomic prediction.

JQ1 Induces DNA Damage and Apoptosis, and Inhibits Tumor Growth in a Patient-Derived Xenograft Model of Cholangiocarcinoma.

Cholangiocarcinoma (CCA) is a fatal disease with a 5-year survival of <30%. For a majority of patients, chemotherapy is the only therapeutic option, and virtually all patients relapse. Gemcitabine is the first-line agent for treatment of CCA. Patients treated with gemcitabine monotherapy survive ∼8 months. Combining this agent with cisplatin increases survival by ∼3 months, but neither regimen produces durable remissions. The molecular etiology of this disease is poorly understood. To facilitate molecular characterization and development of effective therapies for CCA, we established a panel of patient-derived xenograft (PDX) models of CCA. We used two of these models to investigate the antitumor efficacy and mechanism of action of the bromodomain inhibitor JQ1, an agent that has not been evaluated for the treatment of CCA. The data show that JQ1 suppressed the growth of the CCA PDX model CCA2 and demonstrate that growth suppression was concomitant with inhibition of c-Myc protein expression. A second model (CCA1) was JQ1-insensitive, with tumor progression and c-Myc expression unaffected by exposure to this agent. Also selective to CCA2 tumors, JQ1 induced DNA damage and apoptosis and downregulated multiple c-Myc transcriptional targets that regulate cell-cycle progression and DNA repair. These findings suggest that c-Myc inhibition and several of its transcriptional targets may contribute to the mechanism of action of JQ1 in this tumor type. We conclude that BET inhibitors such as JQ1 warrant further investigation for the treatment of CCA. Mol Cancer Ther; 17(1); 107-18. ©2017 AACR.

Ionic Flash NanoPrecipitation (iFNP) for the facile, one-step synthesis of inorganic-organic hybrid nanoparticles in water.

Ionic Flash NanoPrecipitation (iFNP) was evaluated as a novel method for the synthesis of inorganic-organic hybrid nanomaterials and proved to be remarkably effective, fast and practical. To prove the potential of iFNP, various nanostructured GdPO4-based materials of biomedical imaging relevance were easily prepared in a one-step, tunable and highly controlled manner using only water as solvent.

Cervical Spine Clearance in Pediatric Trauma Centers: The Need for Standardization and an Evidence-based Protocol.

BACKGROUND: Cervical spine clearance in the pediatric trauma patient represents a particularly challenging task. Unfortunately, standardized clearance protocols for pediatric cervical clearance are poorly reported in the literature and imaging recommendations demonstrate considerable variability. With the use of a web-based survey, this study aims to define the methods utilized by pediatric trauma centers throughout North America. Specific attention was given to the identification of personnel responsible for cervical spine care, diagnostic imaging modalities used, and the presence or absence of a written pediatric cervical spine clearance protocol. METHODS: A 10-question electronic survey was given to members of the newly formed Pediatric Cervical Spine Study Group, all of whom are active POSNA members. The survey was submitted via the online service SurveyMonkey (https://www.surveymonkey.com/r/7NVVQZR). The survey assessed the respondent's institution demographics, such as trauma level and services primarily responsible for consultation and operative management of cervical spine injuries. In addition, respondents were asked to identify the protocols and primary imaging modality used for cervical spine clearance. Finally, respondents were asked if their institution had a documented cervical spine clearance protocol. RESULTS: Of the 25 separate institutions evaluated, 21 were designated as level 1 trauma centers. Considerable variation was reported with regards to the primary service responsible for cervical spine clearance. General Surgery/Trauma (44%) is most commonly the primary service, followed by a rotating schedule (33%), Neurosugery (11%), and Orthopaedic Surgery (8%). Spine consults tend to be seen most commonly by a rotating schedule of Orthopaedic Surgery and Neurosurgery. The majority of responding institutions utilize computed tomographic imaging (46%) as the primary imaging modality, whereas 42% of hospitals used x-ray primarily. The remaining institutions reported using a combination of x-ray and computed tomographic imaging. Only 46% of institutions utilize a written, standardized pediatric cervical spine clearance protocol. CONCLUSIONS: This study demonstrates a striking variability in the use of personnel, imaging modalities and, most importantly, standardized protocol in the evaluation of the pediatric trauma patient with a potential cervical spine injury. Cervical spine clearance protocols have been shown to decrease the incidence of missed injuries, minimize excessive radiation exposure, decrease the time to collar removal, and lower overall associated costs. It is our opinion that development of a task force or multicenter research protocol that incorporates existing evidence-based literature is the next best step in improving the care of children with cervical spine injuries. LEVEL OF EVIDENCE: Level 4-economic and decision analyses.

This "Ethical Trap" Is for Roboticists, Not Robots: On the Issue of Artificial Agent Ethical Decision-Making.

In this paper we address the question of when a researcher is justified in describing his or her artificial agent as demonstrating ethical decision-making. The paper is motivated by the amount of research being done that attempts to imbue artificial agents with expertise in ethical decision-making. It seems clear that computing systems make decisions, in that they make choices between different options; and there is scholarship in philosophy that addresses the distinction between ethical decision-making and general decision-making. Essentially, the qualitative difference between ethical decisions and general decisions is that ethical decisions must be part of the process of developing ethical expertise within an agent. We use this distinction in examining publicity surrounding a particular experiment in which a simulated robot attempted to safeguard simulated humans from falling into a hole. We conclude that any suggestions that this simulated robot was making ethical decisions were misleading.

Biosensor based on electrospun blended chitosan-poly (vinyl alcohol) nanofibrous enzymatically sensitized membranes for pirimiphos-methyl detection in olive oil.

An ultra-sensitive electrochemical biosensor was successfully developed for rapid detection of pirimiphos-methyl in olive oil, based of genetically-engineered acetylcholinesterase (AChE) immobilization into electrospun chitosan/poly (vinyl alcohol) blend nanofibers. Due to their unique properties such as spatial structure, high porosity, and large surface area, the use of nanofibers allowed improving the biosensor response by two folds. The developed biosensor showed a good performance for detecting pirimiphos-methyl, with a limit of detection of 0.2nM, a concentration much lower than the maximum residue limit allowed set by international regulations (164nM). The biosensor was used for the detection of pirimiphos-methyl in olive oil samples after a simple liquid-liquid extraction, and the recovery rates were close to 100%.

Doubly thermo-responsive copolymers in ionic liquid.

We report the behaviour of thermoresponsive block copolymers of n-butyl acrylate and N-alkyl acrylamides in [C2mim][NTf2]. Poly(N-isopropylacrylamide) exhibits an upper critical solution temperature in [C2mim][NTf2] whereas poly(n-butyl acrylate) has a lower critical solution temperature. Consequently, these polymers exhibit double thermo-responsiveness correlated with the macromolecular structure. Moreover, a switching from micellar to reverse micellar structures was induced by a change in temperature. This property enables the development of reversible shuttles between ionic liquids and water.

Ultra-sensitive biosensor based on genetically engineered acetylcholinesterase immobilized in poly (vinyl alcohol)/Fe-Ni alloy nanocomposite for phosmet detection in olive oil.

An ultra-sensitive screen-printed biosensor was successfully developed for phosmet detection in olive oil, based on a genetically-engineered acetylcholinesterase (AChE) immobilized in a azide-unit water-pendant polyvinyl alcohol (PVA-AWP)/Fe-Ni alloy nanocomposite. Fe-Ni not only allowed amplifying the response current but also lowering the applied potential from 80 mV to 30 mV vs Ag/AgCl. The biosensor showed a very good analytical performance for phosmet detection, with a detection limit of 0.1 nM. This detection limit is lower than the allowable concentrations set by international regulations. In addition to the good reproducibility, operational and storage stability, the developed biosensor was successfully used for the determination of phosmet in olive oil samples without any laborious pre-treatment. The phosmet recovery rate was about 96% after a simple liquid-liquid extraction.

Comparison of methods for the fabrication and the characterization of polymer self-assemblies: what are the important parameters?

The ability to self-assemble was evaluated for a large variety of amphiphilic block copolymers, including poly(ethyleneoxide-b-ε-caprolactone), poly(ethyleneoxide-b-d,l-lactide), poly(ethyleneoxide-b-styrene), poly(ethyleneoxide-b-butadiene) and poly(ethyleneoxide-b-methylmethacrylate). Different methods of formation are discussed, such as cosolvent addition, film hydration or electroformation. The influence of experimental parameters and macromolecular structures on the size and morphology of the final self-assembled structures is investigated and critically compared with the literature. The same process is carried out regarding the characterization of these structures. This analysis demonstrates the great care that should be taken when dealing with such polymeric assemblies. If the morphology of such assemblies can be predicted to some extent by macromolecular parameters like the hydrophilic/hydrophobic balance, those parameters cannot be considered as universal. In addition, external experimental parameters (methods of preparation, use of co-solvent, …) appeared as critical key parameters to obtain a good control over the final structure of such objects, which are very often not at thermodynamic equilibrium but kinetically frozen. A principal component analysis is also proposed, in order to examine the important parameters for forming the self-assemblies. Here again, the hydrophilic/hydrophobic fraction is identified as an important parameter.