Multimode chromatography-based techniques for high purity isolation of extracellular vesicles from human blood plasma.

Extracellular vesicles (EVs) play a pivotal role in various biological pathways, such as immune responses and the progression of diseases, including cancer. However, it is challenging to isolate EVs at high purity from blood plasma and other biofluids due to their low abundance compared to more predominant biomolecular species such as lipoprotein particles and free protein complexes. Ultracentrifugation-based EV isolation, the current gold standard technique, cannot overcome this challenge due to the similar biophysical characteristics of such species. We developed several novel approaches to enrich EVs from plasma while depleting contaminating molecular species using multimode chromatography-based strategies. On average, we identified 716 ± 68 and 1054 ± 35 protein groups in EV isolates from 100 µL of plasma using multimode chromatography- and ultracentrifugation-based techniques, respectively. The developed methods resulted in similar EV isolates purity, providing significant advantages in simplicity, throughput, scalability, and applicability for various downstream analytical and potential clinical applications.

Working like a Dog: Exploring the Role of a Therapy Dog in Clinical Exercise Physiology Practice.

Therapy animals in clinical settings are purported to reduce patients' anxiety, decrease agitated behaviour, serve as social mediators, enhance the social atmosphere, and increase patients' openness towards practitioners. A therapy dog worked alongside her exercise physiologist handler for approximately 1 day/week in a university clinic. The canine and handler functioned as a team, while the handler simultaneously undertook supervision of students. The clinic was open 24 h/week, and no other therapeutic animal was present for any part of the week. We explored, via surveys and interviews, human responses to the dog. The survey comprised 15 statement items regarding the canine's role, behaviour, and acceptability in the clinic, ranked from strongly disagree (-2) to strongly agree (2), followed by an open item inviting participants to follow up interviews. Eleven (11) clinical clients and seven (7) students completed the survey. One client had not encountered the canine; these data were excluded. Four (4) participants from the client sample provided subsequent telephone interviews. All participants identified the canine as well-behaved; no participants considered that she detracted from their exercise sessions. Most participants were equivocal to statements regarding social lubrication and openness to practitioners; only three clients and two students identified that they felt more willing to share health information; three students identified that they felt they could confide more in the canine than in the practitioner. Interviewees' reports were similarly favourable, reinforcing the information obtained from the surveys. Interview transcripts were subject to thematic analysis, which focussed around four key themes: (1) the canine's good behaviour, (2) clients giving permission, and the canine as both (3) a pleasant distraction from the effort of exercise, and (4) nice to have. A therapy dog may enhance some aspects of exercise physiology service delivery.

Inter-regional survey of the New Zealand Pinot noir fermentative sulfur compounds profile.

BACKGROUND: New Zealand Pinot noir is gaining increasing attention both in New Zealand and internationally, becoming the second largest grape variety for both plantings and export. Despite the growing furore around this variety, the current coverage of the volatile chemical profile remains limited, with a lack of information on the fermentative sulfur compounds content in New Zealand Pinot noir wines. RESULTS: Thirty-five Pinot noir wines from three different vintages (i.e. 2016, 2017 and 2018) form five different grape growing regions were analysed for their fermentative sulfur compounds contents. Six fermentative sulfur compounds (i.e. methanethiol, ethanethiol, dimethyl sulfide, carbon disulfide, methionol and benzothiazol) were detected and measured for the first time in New Zealand Pinot noir wines. Their concentrations were compared against previously measured Pinot noir wines from other countries, and some preliminary evidence about inter-regional and ageing effects was obtained. CONCLUSION: The present study reports the first survey of the inter-regional differences in fermentative sulfur compounds contents in 35 New Zealand Pinot noir wines. Preliminary inter-regional and vintage trends prompt further research on the role of these molecules on this wine variety. © 2020 Society of Chemical Industry.

Technologies and Standardization in Research on Extracellular Vesicles.

Extracellular vesicles (EVs) are phospholipid bilayer membrane-enclosed structures containing RNAs, proteins, lipids, metabolites, and other molecules, secreted by various cells into physiological fluids. EV-mediated transfer of biomolecules is a critical component of a variety of physiological and pathological processes. Potential applications of EVs in novel diagnostic and therapeutic strategies have brought increasing attention. However, EV research remains highly challenging due to the inherently complex biogenesis of EVs and their vast heterogeneity in size, composition, and origin. There is a need for the establishment of standardized methods that address EV heterogeneity and sources of pre-analytical and analytical variability in EV studies. Here, we review technologies developed for EV isolation and characterization and discuss paths toward standardization in EV research.

Enhancing patient safety with intelligent intravenous infusion devices: experience in a specialty cardiac hospital.

OBJECTIVE: The study objective was to evaluate patient safety, increase nursing satisfaction, and affect economic factors through implementation of intelligent intravenous (IV) infusion devices in a specialty cardiac hospital. Intelligent IV infusion devices have been shown to decrease medication errors associated with inpatient infusions. METHODS: Intelligent IV infusion device evaluation and drug library creation were conducted by a multidisciplinary team within the hospital. Devices were then implemented into patient care, and the impact was analyzed over a 9-month period. RESULTS: Post-implementation data showed that compliance was approximately 100%. A total of 494 critical catches occurred over the study period, resulting in an estimated annual savings of $7,513,333. End-users became familiar with the new technology and recognized the increase in safety measures and time spent with patients. CONCLUSION: This evaluation suggests that intelligent IV infusion devices resulted in decreased costs and a safer environment for patients.

Operationally realistic validation for prediction of cocoa sensory qualities by high-throughput mass spectrometry.

The potential of analytical chemistry to predict sensory qualities of food materials is a major current theme. Standard practice is cross-validation (CV), where a set of chemical and associated sensory data is partitioned so chemometric models can be developed on training subsets, and validated on held-out subsets. CV demonstrates prediction, but is an unlikely scenario for industrial operations, where concomitant data acquisition for model development and test materials would be unwieldy. We evaluated cocoa materials of diverse provenance, and analyzed on different dates to those used in model development. Liquor extracts were analyzed by flow-injection electrospray-mass spectrometry (FIE-MS), a novel method for sensory quality prediction. FIE-MS enabled prediction of sensory qualities described by trained human panelists. Optimal models came from the Weka data-mining algorithm SimpleLinearRegression, which learns a model for the attribute giving minimal training error, which was (-)-epicatechin. This flavonoid likewise dominated partial least-squares (PLS)-regression models. Refinements of PLS (orthogonal-PLS or orthogonal signal correction) gave poorer generalization to different test sets, as did support vector machines, whose hyperparameters could not be optimized in training to avoid overfitting. In conclusion, if chemometric overfitting is avoided, chemical analysis can predict sensory qualities of food materials under operationally realistic conditions.

Neural correlates of anxiety in healthy volunteers: a voxel-based morphometry study.

Studies have shown that the amygdala, temporal, and prefrontal cortices play a key role in the expression of anxiety. The correlation between gray matter volume of these structures and behavioral anxiety measures was not previously investigated in healthy volunteers. The authors used voxel-based morphometry to assess the relationship between brain regional volume and anxiety. The authors found an inverse correlation between anxiety measures and cortical volume in regions of the limbic system and prefrontal cortex implicated in the pathogenesis of anxiety disorders. The authors suggest that volumetric variability of these regions may have a correlation with the development of an anxious personality trait.

Virulence properties of asymptomatic bacteriuria Escherichia coli.

In asymptomatic bacteriuria (ABU), bacteria colonize the urinary tract without provoking symptoms. Here, we compared the virulence properties of a collection of ABU Escherichia coli strains to cystitis and pyelonephritis strains. Specific urinary tract infection (UTI)-associated virulence genes, hemagglutination characteristics, siderophore production, hemolysis, biofilm formation, and the ability of strains to adhere to and induce cytokine responses in epithelial cells were analyzed. ABU strains were phylogenetically related to strains that cause symptomatic UTI. However, the virulence properties of the ABU strains were variable and dependent on a combination of genotypic and phenotypic factors. Most ABU strains adhered poorly to epithelial cells; however, we also identified a subgroup of strongly adherent strains that were unable to stimulate an epithelial cell IL-6 cytokine response. Poor immune activation may represent one mechanism whereby ABU E. coli evade immune detection after the establishment of bacteriuria.

Integral calculus problem solving: an fMRI investigation.

Only a subset of adults acquires specific advanced mathematical skills, such as integral calculus. The representation of more sophisticated mathematical concepts probably evolved from basic number systems; however its neuroanatomical basis is still unknown. Using fMRI, we investigated the neural basis of integral calculus while healthy participants were engaged in an integration verification task. Solving integrals activated a left-lateralized cortical network including the horizontal intraparietal sulcus, posterior superior parietal lobe, posterior cingulate gyrus, and dorsolateral prefrontal cortex. Our results indicate that solving of more abstract and sophisticated mathematical facts, such as calculus integrals, elicits a pattern of brain activation similar to the cortical network engaged in basic numeric comparison, quantity manipulation, and arithmetic problem solving.

Synthesis, biological evaluation and molecular modelling of N-heterocyclic dipeptide aldehydes as selective calpain inhibitors.

A series of N-heterocyclic dipeptide aldehydes 4-13 have been synthesised and evaluated as inhibitors of ovine calpain 1 (o-CAPN1) and ovine calpain 2 (o-CAPN2). 5-Formyl-pyrrole 9 (IC(50) values of 290 and 25nM against o-CAPN1 and o-CAPN2, respectively) was the most potent and selective o-CAPN2 inhibitor, displaying >11-fold selectivity. The amino acid sequences of o-CAPN1 and o-CAPN2 have been determined. Because of the lack of available structural information on the ovine calpains, in silico homology models of the active site cleft of o-CAPN1 and o-CAPN2 were developed based on human calpain 1 (h-CAPN1) X-ray crystal structure (PDB code 1ZCM). These models were used to rationalise the observed SAR for compounds 4-13 and the selectivity observed for 9. The o-CAPN2 selective inhibitor 9 (CAT0059) was assayed in an in vitro ovine lens culture system and shown to successfully protect the lens from calcium-induced opacification.

Politics on the brain: an FMRI investigation.

We assessed political attitudes using the Implicit Association Test (IAT) in which participants were presented faces and names of well-known Democrat and Republican politicians along with positive and negative words while undergoing functional MRI. We found a significant behavioral IAT effect for the face, but not the name, condition. The fMRI face condition results indicated that ventromedial and anterior prefrontal cortices were activated during political attitude inducement. Amygdala and fusiform gyrus were activated during perceptual processing of familiar faces. Amygdala activation also was associated with measures of strength of emotion. Frontopolar activation was positively correlated with an implicit measure of bias and valence strength (how strongly the participants felt about the politicians), while strength of affiliation with political party was negatively correlated with lateral PFC, lending support to the idea that two distinct but interacting networks-one emphasizing rapid, stereotypic, and emotional associative knowledge and the other emphasizing more deliberative and factual knowledge-cooperate in the processing of politicians. Our findings of ventromedial PFC activation suggests that when processing the associative knowledge concerned with politicians, stereotypic knowledge is activated, but in addition, the anterior prefrontal activations indicate that more elaborative, reflective knowledge about the politician is activated.

Neural correlates of script event knowledge: a neuropsychological study following prefrontal injury.

Scripts sequentially link information about daily activities and event knowledge. Patients have difficulty sequencing script events following lesions of the prefrontal cortex while showing intact access to selective aspects of script knowledge. It has been suggested that the sequencing impairment is due to a deficit in an inhibitory gating mechanisms that usually enables selection of an item from competing alternatives. If this is the case, then an inhibitory task should reveal script processing impairments on a script categorization task that is not normally associated with poor performance following prefrontal damage. To test this hypothesis, we administered a simple untimed classification task and a modified Go/NoGo task in which subjects classified events from social and non-social activities (e.g., read the menu, order the food) and related semantic items (e.g., menu, order) in terms of whether they belonged to a target activity. Participants were patients with lesions of the prefrontal cortex and matched controls. The results showed that damage to the right orbitofrontal cortex was associated with social item classification errors in the simple untimed classification task. In addition, the damage to the right prefrontal cortex was associated with increased response times to respond correctly to Go trials in the modified Go/NoGo task. The data demonstrate that damage to the right orbitofrontal cortex results in impairment in the accessibility of script and semantic representations of social activities. This impairment is exacerbated by an inefficient inhibitory gating mechanism.

Representation of attitudinal knowledge: role of prefrontal cortex, amygdala and parahippocampal gyrus.

It has been proposed that behavior is influenced by representations of different types of knowledge: action representations, event knowledge, attitudes and stereotypes. Attitudes (representations of a concept or object and its emotional evaluation) allow us to respond quickly to a given stimulus. In this study, we explored the representation and inhibition of attitudes. We show that right dorsolateral prefrontal cortex mediates negative attitudes whereas left ventrolateral prefrontal cortex mediates positive attitudes. Parahippocampal regions and amygdala mediate evaluative processing. Furthermore, anxiety modulates right dorsolateral prefrontal activation during negative attitude processing. Inhibition of negative attitudes activates left orbitofrontal cortex: a region that when damaged is associated with socially inappropriate behavior in patients. Inhibition of positive attitudes activates a brain system involving right inferior frontal gyrus and bilateral anterior cingulate. Thus, we show that there are dissociable networks for the representation and inhibition of attitudes.

Psychological structure and neural correlates of event knowledge.

Humans are capable of storing and retrieving sequences of complex structured events. Here we report a study in which we establish the psychological structure of event knowledge and then use parametric event-related functional magnetic resonance imaging to identify its neural correlates. We demonstrate that event knowledge is organized along dissociable dimensions that are reflected in distinctive patterns of neural activation: social valence (amygdala and right orbitofrontal cortex), experience (medial prefrontal cortex) and engagement (left orbitofrontal cortex). Our study affirms the importance and uniqueness of the human prefrontal cortex in representing event knowledge.

Brain activation in processing temporal sequence: an fMRI study.

Structured event complexes (SECs) are stored representations of sequential event knowledge, and represent sequences of activities that have been described elsewhere as scripts or schemas. Previous studies have shown that the prefrontal cortex is involved in temporal sequencing. The present study investigates the involvement of the prefrontal cortex in temporal order and membership judgments of script and category items by using functional magnetic resonance imaging. In this experiment, stimuli were either script events or category items. In experimental trials, subjects classified stimuli according to temporal order or membership category. Results show that the script order task and the chronological order task (relative to their respective memberships tasks) were associated with different patterns of PFC activation. Both order tasks activated the middle frontal gyrus bilaterally; however, script order tasks showed additional activation in right inferior frontal gyrus, and the chronological order tasks in left inferior frontal gyrus. These results suggest that while the middle frontal gyri are activated bilaterally in both script and chronological temporal ordering tasks, there are different, though largely overlapping, neural substrates for script and chronological representations during temporal ordering.

Salicylate accumulation inhibits growth at chilling temperature in Arabidopsis.

The growth of Arabidopsis plants in chilling conditions could be related to their levels of salicylic acid (SA). Plants with the SA hydroxylase NahG transgene grew at similar rates to Col-0 wild types at 23 degrees C, and growth of both genotypes was slowed by transfer to 5 degrees C. However, at 5 degrees C, NahG plants displayed relative growth rates about one-third greater than Col-0, so that by 2 months NahG plants were typically 2.7-fold larger. This resulted primarily from greater cell expansion in NahG rosette leaves. Specific leaf areas and leaf area ratios remained similar in both genotypes. Net assimilation rates were similar in both genotypes at 23 degrees C, but higher in NahG at 5 degrees C. Chlorophyll fluorescence measurements revealed no PSII photodamage in chilled leaves of either genotype. Col-0 shoots at 5 degrees C accumulated SA, particularly in glucosylated form. SA in NahG shoots showed similar tendencies at 5 degrees C, but at greatly depleted levels. Catechol was not detected as a metabolite of the NahG transgene product. We also examined growth and SA levels in SA signaling and metabolism mutants at 5 degrees C. The partially SA-insensitive npr1 mutant displayed growth intermediate between NahG and Col-0, while the SA-deficient eds5 mutant behaved like NahG. In contrast, the cpr1 mutant at 5 degrees C accumulated very high levels of SA and its growth was much more inhibited than wild type. At both temperatures, cpr1 was the only SA-responsive genotype in which oxidative damage (measured as thiobarbituric acid-reactive substances) was significantly different from wild type.

Salicylic acid dependent signaling promotes basal thermotolerance but is not essential for acquired thermotolerance in Arabidopsis thaliana.

Salicylic acid (SA) is reported to protect plants from heat shock (HS), but insufficient is known about its role in thermotolerance or how this relates to SA signaling in pathogen resistance. We tested thermotolerance and expression of pathogenesis-related (PR) and HS proteins (HSPs) in Arabidopsis thaliana genotypes with modified SA signaling: plants with the SA hydroxylase NahG transgene, the nonexpresser of PR proteins (npr1) mutant, and the constitutive expressers of PR proteins (cpr1 and cpr5) mutants. At all growth stages from seeds to 3-week-old plants, we found evidence for SA-dependent signaling in basal thermotolerance (i.e. tolerance of HS without prior heat acclimation). Endogenous SA correlated with basal thermotolerance, with the SA-deficient NahG and SA-accumulating cpr5 genotypes having lowest and highest thermotolerance, respectively. SA promoted thermotolerance during the HS itself and subsequent recovery. Recovery from HS apparently involved an NPR1-dependent pathway but thermotolerance during HS did not. SA reduced electrolyte leakage, indicating that it induced membrane thermoprotection. PR-1 and Hsp17.6 were induced by SA or HS, indicating common factors in pathogen and HS responses. SA-induced Hsp17.6 expression had a different dose-response to PR-1 expression. HS-induced Hsp17.6 protein appeared more slowly in NahG. However, SA only partially induced HSPs. Hsp17.6 induction by HS was more substantial than by SA, and we found no SA effect on Hsp101 expression. All genotypes, including NahG and npr1, were capable of expression of HSPs and acquisition of HS tolerance by prior heat acclimation. Although SA promotes basal thermotolerance, it is not essential for acquired thermotolerance.

Social cognition and the prefrontal cortex.

Social cognitive neuroscience is a rapidly emerging field that utilizes cognitive neuroscientific techniques (e.g., lesion studies, neuroimaging) to address concepts traditionally in the social psychological realm (e.g., attitudes, stereotypes). The purpose of this article is to review published neuroscientific and neuropsychological research into social cognition. The author focuses on the role of the prefrontal cortex in social behavior and presents a framework that provides cohesion of this research. The article proposes that this framework will be useful in guiding future social cognitive neuroscientific research.

Assessment of antimicrobial activity of hydrophilic betaines in osmotically stressed bacteria.

A series of hydrophilic aromatic and semi-aromatic betaines related to trigonelline was synthesized and tested for antimicrobial activity. 4-Methylthiazolium betaine was the only one that showed significant antibacterial activity towards Escherichia coli under hyperosmotic conditions. None of the tested betaines showed any evidence of osmoprotection or urea protection.