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Organelle heterogeneity and inter-organelle contacts within a single cell contribute to the limited sensitivity of current organelle separation techniques, thus hindering organelle subpopulation characterization. Here, we use direct current insulator-based dielectrophoresis (DC-iDEP) as an unbiased separation method and demonstrate its capability by identifying distinct distribution patterns of insulin vesicles from INS-1E insulinoma cells. A multiple voltage DC-iDEP strategy with increased range and sensitivity has been applied, and a differentiation factor (ratio of electrokinetic to dielectrophoretic mobility) has been used to characterize features of insulin vesicle distribution patterns. We observed a significant difference in the distribution pattern of insulin vesicles isolated from glucose-stimulated cells relative to unstimulated cells, in accordance with maturation of vesicles upon glucose stimulation. We interpret the difference in distribution pattern to be indicative of high-resolution separation of vesicle subpopulations. DC-iDEP provides a path for future characterization of subtle biochemical differences of organelle subpopulations within any biological system.
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Electroforesis , Insulina , Vesículas Secretoras , Electroforesis/métodos , Insulina/metabolismo , Vesículas Secretoras/metabolismo , Vesículas Secretoras/química , Animales , Ratas , Línea Celular Tumoral , Glucosa/metabolismoRESUMEN
Background: The World Health Organization seeks to eliminate viral hepatitis as a public health threat by 2030. This review and meta-analysis aims to evaluate the effectiveness of programs for hepatitis B and C testing and treatment in community pharmacies. Methods: Medline, Embase, Cochrane CENTRAL, and Global Health were searched from database inception until 12 November 2023. Comparative and single arm intervention studies were eligible for inclusion if they assessed delivery of any of the following interventions for hepatitis B or C in pharmacies: (1) pre-testing risk assessment, (2) testing, (3) pre-treatment assessment or (4) treatment. Primary outcomes were proportions testing positive and reaching each stage in the cascade. Random effects meta-analysis was used to estimate pooled proportions stratified by recruitment strategy and setting where possible; other results were synthesised narratively. This study was pre-registered (PROSPERO: CRD42022324218). Findings: Twenty-seven studies (4 comparative, 23 single arm) were included, of which 26 reported hepatitis C outcomes and four reported hepatitis B outcomes. History of injecting drug use was the most identified risk factor from pre-testing risk assessments. The pooled proportion hepatitis C antibody positive from of 19 studies testing 5096 participants was 16.6% (95% CI 11.0%-23.0%; heterogeneity I2 = 96.6%). The pooled proportion antibody positive was significantly higher when testing targeted people with specified risk factors (32.5%, 95% CI 24.8%-40.6%; heterogeneity I2 = 82.4%) compared with non-targeted or other recruitment methods 4.0% (95% CI 2.1%-6.5%; heterogeneity I2 = 83.5%). Meta-analysis of 14 studies with 813 participants eligible for pre-treatment assessment showed pooled attendance rates were significantly higher in pharmacies (92.7%, 95% CI 79.1%-99.9%; heterogeneity I2 = 72.4%) compared with referral to non-pharmacy settings (53.5%, 95% CI 36.5%-70.1%; heterogeneity I2 = 92.3%). The pooled proportion initiating treatment was 85.6% (95% CI 74.8%-94.3%; heterogeneity I2 = 75.1%). This did not differ significantly between pharmacy and non-pharmacy settings. Interpretation: These findings add pharmacies to the growing evidence supporting community-based testing and treatment for hepatitis C. Few comparative studies and high degrees of statistical heterogeneity were important limitations. Hepatitis B care in pharmacies presents an opportunity for future research. Funding: None.
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Sleep is often compromised in female athletes, and the monitoring of female athletes' sleep is an important preventative and educational tool. With self-reporting of sleep common practice for athletes as part of a daily wellness assessment, there is a need to understand whether sleep indices are being reported accurately, and thus whether self-report data are useful. This study aimed to compare the agreement between self-reported and actigraphy reported sleep duration in female football players, with the intention of informing best practice for athlete monitoring. Twenty-two female footballers (mean age 19.5 ± 1.3 years) provided a daily self-report across 7 days, whilst also wearing an actigraph across the same testing period. Agreement between the two measures was assessed using Bland-Altman limits of agreement, with acceptable limits of agreement defined as <30 minutes. Results showed evident disagreement between the two methods, with a mean bias of -0.54 (32 min, 95% CI -0.66 to -0.43) and a potential disagreement range of over 2 h (Lower 95% limits of agreement -1.49 to upper 95% limits of agreement 0.40). Coaches using self-reported sleep durations as a monitoring tool for female footballers should interpret the results with caution and be aware of the potential for inaccuracies in this measure. As such, where possible, coaches should consider other methods of sleep monitoring, rather than solely relying on a self-report, to ensure they are operating with optimal practice within situational constraints.
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This case presents a known complication of particulate synovitis granuloma associated with a first metatarsophalangeal joint silastic implant. However, the degree of soft tissue granuloma enlargement is quite unique in size and its proliferative effect-invading the medulla cavity and infiltrating the outer cortex of bone. This case study aims to demonstrate its clinical presentation, imaging investigations, surgical excision and histopathology findings. The learning points emphasised within this manuscript draw attention to the procedure selection for a silastic implant, as well as its proposed mode of action and various potential associated complications. Surgery was based on careful analysis of overall function, prior surgery conducted and patient expectations to achieve a shared decision-making process.
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Prótesis Articulares , Articulación Metatarsofalángica , Sinovitis , Humanos , Prótesis Articulares/efectos adversos , Articulación Metatarsofalángica/diagnóstico por imagen , Articulación Metatarsofalángica/cirugía , Articulación Metatarsofalángica/patología , Sinovitis/etiología , Granuloma/patologíaRESUMEN
Nephrogenic adenoma (NA) is a rare, benign lesion of the urinary tract that is induced by chronic irritation or injury to the urinary tract. Ureteral nephrogenic adenoma arising from both ureters is an exceptionally rare condition. We report an unusual case of a 73-year-old male who presented with a several-month history of recurrent UTI-like symptoms. Subsequent imaging showed bilateral hydronephrosis and ureteral wall thickening. A retrograde ureteroscopy revealed several papillary masses filling the lumens of both ureters. Ureteroscopic biopsies revealed NA in both ureters.
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Determining the physical and chemical properties of biologically important particles such as cells, organelles, viruses, exosomes, complexes, nucleotides, and proteins is needed to understand their function. These properties are determined with common analytical tools (mass spectrometry, cryo-EM, NMR, various spectroscopies, nucleotide sequencing, etc.) whose function can be improved when samples are pure and concentrated. Separations science plays a central role in conditioning samples, ranging from low-resolution benchtop operations like precipitations or extractions to higher-resolution chromatography and electrophoresis. In the last two decades, gradient insulator-based dielectrophoresis (g-iDEP) has emerged as a high-resolution separation technique capable of highly selective enrichment of cells, viruses, exosomes, and proteins. Specific evidence has been shown that pure homogeneous and concentrated fractions of cells and exosomes can be generated from complex mixtures. However, recovering those fractions for analysis has not been developed, limiting the technique to an analytical rather than a preparative one. Here, a finite element analysis was undertaken to identify geometries and operational parameters to efficiently remove the enriched fraction while retaining maximum concentration and providing total mass transfer. Geometric factors (e.g., side channel width and distance from the gradient-inducing gap) were studied, along with the addition of a second inlet side channel. Two flow-generating mechanisms-electroosmosis and hydrostatic pressure-were evaluated for semi-optimized device designs, including a comparison of the one- and two-inlet designs. Simulations indicate effectively one hundred percent mass transfer and a concentration increase by an order of magnitude for several device configurations and operational parameters.
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Electroósmosis , Técnicas Analíticas Microfluídicas , Electroforesis/métodos , Electroósmosis/métodos , Dispositivos Laboratorio en un ChipRESUMEN
Background: Basal cell carcinoma (BCC) is one of the most common malignancies in the world. The frequency of histopathological subtypes and the distribution on the body of BCC has been well documented. Less has been written on the nature of secondary tumours. The genetics of BCC is starting to be understood, particularly with the advent of newer medical treatments (hedgehog inhibitors). Objectives: To determine if primary basal cell carcinoma histopathological subtype predicts secondary tumour subtype, as well as their anatomical distribution. Methods: A retrospective case series of patients over the age of 18 was performed from 2009 to 2014, with at least two separate diagnoses of BCC. Results: In 394 identified patients, a total of 1355 BCCs arose in the cohort over the 6-year study period. The number of secondary BCCs per patient ranged from 2 to 19 tumours. Nodular BCC was the most likely to reoccur in secondary tumours (53.3%), followed by mixed subtypes (45.7%). Conclusions: Within our study, we did find a predisposition for secondary BCCs to be of the same histopathological subtype as the primary, particularly with respect to nodular and mixed tumours. Furthermore, we found that secondary tumours were also more likely to occur on the same anatomical site as the primary tumour. We are only just beginning to under the genetic mutations involved in subtype formation.
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Therapeutics at or close to the nanoscale, such as liposomal irinotecan, offer significant promise for the treatment of solid tumors. Their potential advantage over the unencapsulated or free form of the drug is due in part to their altered biodistribution. For slow and sustained release, significant optimization of formulation is needed to achieve the required level of stability and allow long-term storage of the drug product. Gradient-based liposomal formulation of camptothecins such as irinotecan poses unique challenges owing to the camptothecin- and acid-catalyzed hydrolysis of phospholipid esters in the inner monolayer of the liposomal membrane. We demonstrated that a narrow set of conditions related to the external pH, temperature, intraliposomal concentration, identity of the drug-trapping agent, physical form of the drug inside the liposomes, and final drug load have a marked impact on the stability of the liposome phospholipid membrane. The physical form of the drug inside the liposome was shown to be an insoluble gel with an irinotecan-to-sulfate ratio approximating 1:1, reducing the potential for irinotecan-catalyzed phospholipid hydrolysis in the internal phospholipid monolayer. As a result of this work, a stable and active liposome formulation has been developed that maintains phospholipid chemical stability following long-term storage at 2-8°C.
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Liposomas , Fosfolípidos , Irinotecán , Estabilidad de Medicamentos , Distribución Tisular , Camptotecina , CatálisisRESUMEN
The development of elegant and numerous microfluidic manipulations has enabled significant advances in the processing of small volume samples and the detection of minute amounts of biomaterials. Effective isolation of single cells in a defined volume as well as manipulations of complex bioparticle or biomolecule mixtures allows for the utilization of information-rich detection methods including mass spectrometry, electron microscopy imaging, and amplification/sequencing. The art and science of translating biosamples from microfluidic platforms to highly advanced, information-rich detection system is the focus of this review, where we term the translation between the microfluidics elements to the external world "off-chipping." When presented with the challenge of presenting sub-nanoliter volumes of manipulated sample to a detection scheme, several delivery techniques have been developed for effective analysis. These techniques include spraying (electrospray, nano-electrospray, pneumatic), meniscus-defined volumes (droplets, plugs), constrained volumes (narrow channels, containers), and phase changes (deposition, freezing). Each technique has been proven effective in delivering highly defined samples from microfluidic systems to the detection elements. This review organizes and presents selective publications that illustrate the advancements of these delivery techniques with respect to the type of sample analyzed, while introducing each strategy and providing historical perspective. The publications highlighted in this review were chosen due to their significance and relevance in the development of their respective off-chip technique. This review highlights advancements of delivery methods off a microfluidic chip for additional information rich detection schemes.
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Técnicas Analíticas Microfluídicas , Microfluídica , Espectrometría de Masas , Técnicas Analíticas Microfluídicas/métodos , Microfluídica/métodos , Análisis de Secuencia por Matrices de OligonucleótidosRESUMEN
Triplex gene editing relies on binding a stable peptide nucleic acid (PNA) sequence to a chromosomal target, which alters the helical structure of DNA to stimulate site-specific recombination with a single-strand DNA (ssDNA) donor template and elicits gene correction. Here, we assessed whether the codelivery of PNA and donor template encapsulated in Poly Lactic-co-Glycolic Acid (PLGA)-based nanoparticles can correct sickle cell disease and x-linked severe combined immunodeficiency. However, through this process we have identified a false-positive PCR artifact due to the intrinsic capability of PNAs to aggregate with ssDNA donor templates. Here, we show that the combination of PNA and donor templates but not either agent alone results in different degrees of aggregation that result in varying but highly reproducible levels of false-positive signal. We have identified this phenomenon in vitro and confirmed that the PNA sequences producing the highest supposed correction in vitro are not active in vivo in both disease models, which highlights the importance of interrogating and eliminating carryover of ssDNA donor templates in assessing various gene editing technologies such as PNA-mediated gene editing.
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Edición Génica/métodos , Anemia de Células Falciformes/genética , Animales , Reacciones Falso Positivas , Subunidad gamma Común de Receptores de Interleucina/genética , Ratones SCID , Técnicas de Sonda Molecular , Ácidos Nucleicos de Péptidos , Copolímero de Ácido Poliláctico-Ácido PoliglicólicoRESUMEN
OBJECTIVES: To investigate the efficacy of heat acclimation (HA) in the young (YEX) and elderly (EEX) following exercise-HA, and the elderly utilising post-exercise hot water immersion HA (EHWI). DESIGN: Cross-sectional study. METHOD: Twenty-six participants (YEX: nâ¯=â¯11 aged 22⯱â¯2â¯years, EEX:nâ¯=â¯8 aged 68⯱â¯3â¯years, EHWI: nâ¯=â¯7 aged 73⯱â¯3â¯years) completed two pre-/post-tests, separated by five intervention days. YEX and EEX exercised in hot conditions to raise rectal temperature (Trec) ≥38.5⯰C within 60â¯min, with this increase maintained for a further 60â¯min. EHWI completed 30â¯min of cycling in temperate conditions, then 30â¯min of HWI (40⯰C), followed by 30â¯min seated blanket wrap. Pre- and post-testing comprised 30â¯min rest, followed by 30â¯min of cycling exercise (3.5â¯W·kg-1 Hprod), and a six-minute walk test (6MWT), all in 35⯰C, 50% RH. RESULTS: The HA protocols did not elicit different mean heart rate (HR), Trec, and duration Trecâ¯≥â¯38.5⯰C (pâ¯>â¯0.05) between YEX, EEX, and EHWI groups. Resting Trec, peak skin temperature, systolic and mean arterial pressure, perceived exertion and thermal sensation decreased, and 6MWT distance increased pre- to post-HA (pâ¯<â¯0.05), with no difference between groups. YEX also demonstrated a reduction in resting HR (pâ¯<â¯0.05). No change was observed in peak Trec or HR, vascular conductance, sweat rate, or thermal comfort in any group (pâ¯>â¯0.05). CONCLUSIONS: Irrespective of age or intervention, HA induced thermoregulatory, perceptual and exercise performance improvements. Both exercise-HA (EEX), and post-exercise HWI (EHWI) are considered viable interventions to prepare the elderly for heat stress.
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Aclimatación , Envejecimiento/fisiología , Ejercicio Físico/fisiología , Respuesta al Choque Térmico , Calor , Inmersión , Anciano , Presión Sanguínea , Estudios Transversales , Femenino , Frecuencia Cardíaca , Trastornos de Estrés por Calor/prevención & control , Humanos , Masculino , Percepción/fisiología , Esfuerzo Físico/fisiología , Descanso , Temperatura Cutánea , Sudoración , Sensación Térmica , Adulto JovenRESUMEN
NEW FINDINGS: What is the central question of this study? Fire service instructors are frequently exposed to live fire scenarios, representing the most extreme chronic occupational heat exposure. These individuals report a series of unique health issues. We sought to identify whether the number of exposures completed was associated with inflammatory and immunological markers and symptoms of ill health. What is the main finding and its importance? Fire service instructors exhibit greater levels of inflammatory markers in comparison to firefighters. The number of exposures to fire is positively related to the prevalence of ill health and inflammation. Implementation of a proposed limit of nine exposures per month might be appropriate to minimize health issues. ABSTRACT: Fire Service Instructors (FSIs) experience â¼10 times more fire exposures than firefighters (FFs), and the increased physiological stress from this potentially puts them at risk of ill health and future cardiac events. The aim of the study was to establish whether FSIs exhibit elevated biomarkers associated with cardiac event risk, identify whether FSIs experience systemic inflammation linked to the frequency of fire exposure and evaluate a proposed exposure limit of nine exposures per month. Blood samples were collected from 110 Fire Service personnel (mean ± SD, age,44 ± 7 years; height, 178.1 ± 7.1 cm; and body mass, 84.3 ± 12.0 kg; FSIs n = 53 and FFs n = 57) for biomarker analysis. Work history details were collected from all participants. Participants with biomarker concentrations above healthy reference ranges were classified as being 'at risk'. The neutrophil-to-lymphocyte ratio, platelet count, cardiac troponin T, interleukin (IL)-6, IL-1ß, C-reactive protein and immunoglobulin G were greater in FSIs than in FFs (P < 0.05). Multiple regression analysis revealed that 18.8% of IL-6, 24.9% of IL-1ß, 29.2% of C-reactive protein and 10.9% of immunoglobulin G variance could be explained by the number of exposures to heat per month. Odds ratios revealed that those FSIs above the nine per month exposure limit were six to 12 times more likely to be classified as 'at risk' and were 16 times more likely to experience symptoms of ill health. Increased cytokine levels suggest that FSIs experience systemic inflammation, which is related to symptoms of ill health. We propose that an exposure limit could reduce the prevalence of these biomarker risk factors and ill health.
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Calor/efectos adversos , Inflamación/sangre , Exposición Profesional , Estrés Fisiológico/fisiología , Adulto , Biomarcadores/sangre , Proteína C-Reactiva/metabolismo , Bomberos , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Proteins are perhaps the most important yet frustratingly complicated and difficult class of compounds to analyze, manipulate, and use. One very attractive option to characterize and differentially concentrate proteins is dielectrophoresis, but according to accepted theory, the force on smaller particles the size of proteins is too low to overcome diffusive action. Here, three model proteins, immunoglobulin G, α-chymotrypsinogen A, and lysozyme, are shown to generate forces much larger than predicted by established theory are more consistent with new theoretical constructs, which include the dipole moment and interfacial polarizability. The forces exerted on the proteins are quantitatively measured against well-established electrophoretic and diffusive processes and differ for each. These forces are orders of magnitude larger than previously predicted and enable the selective isolation and concentration of proteins consistent with an extremely high-resolution separation and concentration system based on the higher-order electric properties. The separations occur over a small footprint, happen quickly, and can be made in series or parallel (and in any order) on simple devices.
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Quimotripsinógeno/análisis , Inmunoglobulina G/análisis , Muramidasa/análisis , Animales , Pollos , Clara de Huevo/análisis , Electroforesis , Muramidasa/metabolismoRESUMEN
BACKGROUND: Assessment of pure polysaccharide response to the 23-valent pneumococcal polysaccharide vaccine (PPV23) can be biased by previous exposure to the conjugate vaccine (PCV). We applied pre-analytical modification to the existing ELISA by pre-incubating serum with PCV. METHODS: PCV-adsorbed and non-adsorbed sera were prepared before measuring the concentration of anti-pneumococcal capsular polysaccharide (PCP) IgG antibodies by the whole pneumococcal ELISA. Paired pre and post-pneumococcal vaccination sera from 73 subjects were analyzed and the baseline anti-PCP IgG for each sample was subtracted from the post-vaccination value to measure vaccine responses. Absolute change in titers and fold changes were then compared between both methods. RESULTS: In the PCV-vaccinated group (n = 28), pre-adsorption with PCV significantly reduced the vaccine responses compared to non-adsorbed sera [median increase in anti-PCP titers: 27.55 mg/l and 45.98 mg/l, respectively]. In addition, the median fold change dropped significantly from 3.026 to 2.313. In PPV23-vaccinated immunocompetent subjects (n = 28) there was a significant difference in anti-PCP responses with PCV adsorption [median values: 73.71 mg/l without and 51.04 mg/l with adsorption]. All the antibody deficiency patients (n = 17) displayed poor PPV23 responses. Although PPV23 responsiveness was not statistically different between both methods, we have observed a trend for lower anti-PCP IgG titers in PCV-adsorbed sera compared to non-adsorbed ones. Serotype-specific IgG analysis using a multiplexed bead-based immunoassay performed on 10 paired samples confirmed that the adsorption observed is specific to PCV serotypes. CONCLUSION: Pre-analytical modification to the conventional ELISA by removing the PCV-specific serotypes may differentiate true polysaccharide response from recall response induced by previous PCV vaccination.
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Anticuerpos Antibacterianos/sangre , Ensayo de Inmunoadsorción Enzimática , Inmunoglobulina G/sangre , Vacunas Neumococicas/uso terapéutico , Pruebas Serológicas , Vacunación , Biomarcadores/sangre , Humanos , Inmunización Secundaria , Inmunogenicidad Vacunal , Vacunas Neumococicas/inmunología , Valor Predictivo de las Pruebas , Prueba de Estudio Conceptual , Estudios Retrospectivos , Factores de Tiempo , Resultado del TratamientoRESUMEN
The ability to selectively move and trap proteins is core to their effective use as building blocks and for their characterization. Analytical and preparative strategies for proteins have been pursued and modeled for nearly a hundred years, with great advances and success. Core to all of these studies is the separation, isolation, purification, and concentration of pure homogeneous fractions of a specific protein in solution. Processes to accomplish this useful solution include biphasic equilibrium (chromatographies, extractions), mechanical, bulk property, chemical equilibria, and molecular recognition. Ultimately, the goal of all of these is to physically remove all non-like protein molecules-to the finest detail: all atoms in the full three-dimensional structure being identical down the chemical bond and bulk structure chirality. One strategy which has not been effectively pursued is exploiting the higher order subtle electrical properties of the protein-solvent system. The advent of microfluidic systems has enabled the use of very high electric fields and well-defined gradients such that extremely high resolution separations of protein mixtures are possible. These advances and recognition of these capabilities have caused a re-evaluation of the underlying theoretical models and they were found to be inadequate. New theoretical descriptions are being considered which align more closely to the total forces present and the subtlety of differences between similar proteins. These are focused on the interfacial area between the protein and hydrating solvent molecules, as opposed to the macroscale assumptions of homogeneous solutions and particles. This critical review examines all data which has been published that place proteins in electric field gradients which induce collection of those proteins, demonstrating a force greater than dispersive effects or countering forces. Evolving theoretical constructs are presented and discussed, and a general estimate of future capabilities using the higher order effects and the high fields and precise gradients of microfluidic systems is discussed. Graphical abstract.
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Electroforesis/métodos , Proteínas/aislamiento & purificación , Modelos Teóricos , Proteínas/químicaRESUMEN
Salmonella is an important pathogen and is a world-wide threat to food safety and public health. Surveillance of serotypes and fundamental biological and biochemical studies are supported by a wide variety of established and emerging bioanalytical techniques. These include classic serotyping based on the Kauffmann-White nomenclature and the emerging whole genome sequencing strategy. Another emerging strategy is native whole cell biophysical characterization which has yet to be applied to Salmonella. However, this technique has been shown to provide high resolution differentiation of serotypes with several other paired strains of other microbes and pathogens. To demonstrate that biophysical characterization might be useful for Salmonella serotyping, the closely related strains sv. Cubana and sv. Poona were chosen for study. These two serovars were subjected to biophysical measurements on a dielectrophoresis-based microfluidic device that generated full differentiation of the unlabeled and native cells. They were differentiated by the ratio of electrophoretic (EP) to dielectrophoretic (DEP) mobilities. This differentiation factor is 2.7 ± 0.3 × 1010 V/m2 for sv. Cubana, versus 2.2 ± 0.3 × 1010 V/m2 for sv. Poona. This work shows for the first time the differentiation, concentration, and characterization of the Salmonella serotypes by exploiting their biophysical properties. It may lead to a less expensive and more decentralized new tool and method for microbiologists, complimenting and working in parallel with other characterization methods.
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International competition inevitably presents logistical challenges for athletes. Events such as the Tokyo 2020 Olympic Games require further consideration given historical climate data suggest athletes will experience significant heat stress. Given the expected climate, athletes face major challenges to health and performance. With this in mind, heat alleviation strategies should be a fundamental consideration. This review provides a focused perspective of the relevant literature describing how practitioners can structure male and female athlete preparations for performance in hot, humid conditions. Whilst scientific literature commonly describes experimental work, with a primary focus on maximizing magnitudes of adaptive responses, this may sacrifice ecological validity, particularly for athletes whom must balance logistical considerations aligned with integrating environmental preparation around training, tapering and travel plans. Additionally, opportunities for sophisticated interventions may not be possible in the constrained environment of the athlete village or event arenas. This review therefore takes knowledge gained from robust experimental work, interprets it and provides direction on how practitioners/coaches can optimize their athletes' heat alleviation strategies. This review identifies two distinct heat alleviation themes that should be considered to form an individualized strategy for the athlete to enhance thermoregulatory/performance physiology. First, chronic heat alleviation techniques are outlined, these describe interventions such as heat acclimation, which are implemented pre, during and post-training to prepare for the increased heat stress. Second, acute heat alleviation techniques that are implemented immediately prior to, and sometimes during the event are discussed. Abbreviations: CWI: Cold water immersion; HA: Heat acclimation; HR: Heart rate; HSP: Heat shock protein; HWI: Hot water immersion; LTHA: Long-term heat acclimation; MTHA: Medium-term heat acclimation; ODHA: Once-daily heat acclimation; RH: Relative humidity; RPE: Rating of perceived exertion; STHA: Short-term heat acclimation; TCORE: Core temperature; TDHA: Twice-daily heat acclimation; TS: Thermal sensation; TSKIN: Skin temperature; VÌO2max: Maximal oxygen uptake; WGBT: Wet bulb globe temperature.