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1.
Biochem J ; 479(15): 1609-1619, 2022 08 12.
Artigo em Inglês | MEDLINE | ID: mdl-35851603

RESUMO

Human BK channels are large voltage and Ca2+-activated K+ channels, involved in several important functions within the body. The core channel is a tetramer of α subunits, and its function is modulated by the presence of ß and γ accessory subunits. BK channels composed of α subunits, as well as BK channels composed of α and ß1 subunits, were successfully solubilised from HEK cells with styrene maleic acid (SMA) polymer and purified by nickel affinity chromatography. Native SMA-PAGE analysis of the purified proteins showed the α subunits were extracted as a tetramer. In the presence of ß1 subunits, they were co-extracted with the α subunits as a heteromeric complex. Purified SMA lipid particles (SMALPs) containing BK channel could be inserted into planar lipid bilayers (PLB) and single channel currents recorded, showing a high conductance (≈260 pS), as expected. The open probability was increased in the presence of co-purified ß1 subunits. However, voltage-dependent gating of the channel was restricted. In conclusion, we have demonstrated that SMA can be used to effectively extract and purify large, complex, human ion channels, from low expressing sources. That these large channels can be incorporated into PLB from SMALPs and display voltage-dependent channel activity. However, the SMA appears to reduce the voltage dependent gating of the channels.


Assuntos
Ativação do Canal Iônico , Canais de Potássio Ativados por Cálcio de Condutância Alta , Humanos , Subunidades alfa do Canal de Potássio Ativado por Cálcio de Condutância Alta/genética , Subunidades alfa do Canal de Potássio Ativado por Cálcio de Condutância Alta/metabolismo , Canais de Potássio Ativados por Cálcio de Condutância Alta/metabolismo
2.
PLoS Biol ; 15(9): e2001655, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28902870

RESUMO

Lowered insulin/insulin-like growth factor (IGF) signaling (IIS) can extend healthy lifespan in worms, flies, and mice, but it can also have adverse effects (the "insulin paradox"). Chronic, moderately lowered IIS rescues age-related decline in neurotransmission through the Drosophila giant fiber system (GFS), a simple escape response neuronal circuit, by increasing targeting of the gap junctional protein innexin shaking-B to gap junctions (GJs). Endosomal recycling of GJs was also stimulated in cultured human cells when IIS was reduced. Furthermore, increasing the activity of the recycling small guanosine triphosphatases (GTPases) Rab4 or Rab11 was sufficient to maintain GJs upon elevated IIS in cultured human cells and in flies, and to rescue age-related loss of GJs and of GFS function. Lowered IIS thus elevates endosomal recycling of GJs in neurons and other cell types, pointing to a cellular mechanism for therapeutic intervention into aging-related neuronal disorders.


Assuntos
Envelhecimento/fisiologia , Drosophila/fisiologia , Insulina/metabolismo , Somatomedinas/metabolismo , Transmissão Sináptica , Animais , Conexinas/metabolismo , Reação de Fuga/fisiologia , Feminino , Junções Comunicantes/fisiologia , Masculino , Proteínas rab de Ligação ao GTP/metabolismo
3.
J Hand Ther ; 33(2): 180-187, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32279878

RESUMO

STUDY DESIGN: Scoping review. INTRODUCTION: With the recent advances in technologies, interactive wearable technologies including inertial motion sensors and e-textiles are emerging in the field of rehabilitation to monitor and provide feedback and therapy remotely. PURPOSE OF THE STUDY: This review article focuses on inertial measurement unit motion sensor and e-textiles-based technologies and proposes approaches to augment these interactive wearable technologies. METHODS: We conducted a comprehensive search of relevant electronic databases (eg, PubMed, the Cumulative Index to Nursing and Allied Health Literature, Embase, PsycINFO, The Cochrane Central Register of Controlled Trial, and the Physiotherapy Evidence Database). The scoping review included all study designs. RESULTS: Currently, there are a numerous research groups and companies investigating inertial motion sensors and e-textiles-based interactive wearable technologies. However, translation of these technologies to the clinic would need further research to increase ease of use and improve clinical validity of the outcomes of these technologies. DISCUSSION: The current review discusses the limitations of the interactive wearable technologies such as, limited clinical utility, bulky equipment, difficulty in setting up equipment inertial motion sensors and e-textiles. CONCLUSION: There is tremendous potential for interactive wearable technologies in rehabilitation. With the evolution of cloud computing, interactive wearable systems can remotely provide intervention and monitor patient progress using models of telerehabilitation. This will revolutionize the delivery of rehabilitation and make rehabilitation more accessible and affordable to millions of individuals.


Assuntos
Eletromiografia/instrumentação , Monitorização Fisiológica/instrumentação , Movimento (Física) , Modalidades de Fisioterapia/instrumentação , Extremidade Superior , Dispositivos Eletrônicos Vestíveis , Humanos
4.
Methods ; 147: 206-212, 2018 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-29526775

RESUMO

There are a number of methods of investigating the function of recombinant proteins such as ion channels. However, after channel purification there are few methods to guarantee that the protein still functions. For ion channels, reconstituting back into planar lipid bilayers and demonstrating preserved function is a convenient and trusted method. It is cell free and even inaccessible, intracellular ion channels can be studied. We have used this method to study the function of recombinant channels of known subunit composition and have found it convenient for investigating the mode of action of ion channel modulators.


Assuntos
Canais de Potássio Ativados por Cálcio de Condutância Alta/fisiologia , Bicamadas Lipídicas/metabolismo , Proteínas Recombinantes/metabolismo , Células HEK293 , Humanos , Ativação do Canal Iônico , Razão Sinal-Ruído
5.
Sensors (Basel) ; 19(5)2019 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-30823373

RESUMO

Rehabilitation following knee injury or surgery is critical for recovery of function and independence. However, patient non-adherence remains a significant barrier to success. Remote rehabilitation using mobile health (mHealth) technologies have potential for improving adherence to and execution of home exercise. We developed a remote rehabilitation management system combining two wireless inertial measurement units (IMUs) with an interactive mobile application and a web-based clinician portal (interACTION). However, in order to translate interACTION into the clinical setting, it was first necessary to verify the efficacy of measuring knee motion during rehabilitation exercises for physical therapy and determine if visual feedback significantly improves the participant's ability to perform the exercises correctly. Therefore, the aim of this study was to verify the accuracy of the IMU-based knee angle measurement system during three common physical therapy exercises, quantify the effect of visual feedback on exercise performance, and understand the qualitative experience of the user interface through survey data. A convenience sample of ten healthy control participants were recruited for an IRB-approved protocol. Using the interACTION application in a controlled laboratory environment, participants performed ten repetitions of three knee rehabilitation exercises: heel slides, short arc quadriceps contractions, and sit-to-stand. The heel slide exercise was completed without feedback from the mobile application, then all exercises were performed with visual feedback. Exercises were recorded simultaneously by the IMU motion tracking sensors and a video-based motion tracking system. Validation showed moderate to good agreement between the two systems for all exercises and accuracy was within three degrees. Based on custom usability survey results, interACTION was well received. Overall, this study demonstrated the potential of interACTION to measure range of motion during rehabilitation exercises for physical therapy and visual feedback significantly improved the participant's ability to perform the exercises correctly.


Assuntos
Articulação do Joelho/fisiopatologia , Sistemas de Identificação de Pacientes/métodos , Reabilitação/instrumentação , Reabilitação/métodos , Telerreabilitação/instrumentação , Telerreabilitação/métodos , Tecnologia sem Fio/instrumentação , Adulto , Exercício Físico/fisiologia , Terapia por Exercício/instrumentação , Terapia por Exercício/métodos , Retroalimentação , Feminino , Humanos , Masculino , Aplicativos Móveis , Amplitude de Movimento Articular/fisiologia , Adulto Jovem
6.
Breast Cancer Res ; 19(1): 35, 2017 03 24.
Artigo em Inglês | MEDLINE | ID: mdl-28340615

RESUMO

BACKGROUND: Psychological stress increases the circulating levels of the stress hormones cortisol and norepinephrine (NE). Chronic exposure to elevated stress hormones has been linked to a reduced response to chemotherapy through induction of DNA damage. We hypothesize that stress hormone signalling may induce DNA damage through the production of reactive oxygen species (ROS)/reactive nitrogen species (RNS) and interference in DNA repair processes, promoting tumourigenesis. METHODS: Breast cancer cell lines were incubated with physiological levels of cortisol and NE in the presence and absence of receptor antagonists and inducible nitric oxide synthase (iNOS) inhibitors and DNA damage measured using phosphorylated γ-H2AX. The rate of DNA repair was measured using comet assays and electrochemical sensors were used to detect ROS/RNS in the cell lysates from cells exposed to stress hormones. A syngeneic mouse model was used to assess the presence of iNOS in mammary tumours in stressed versus control animals and expression of iNOS was examined using western blotting and qRT-PCR. RESULTS: Acute exposure to cortisol and NE significantly increased levels of ROS/RNS and DNA damage and this effect was diminished in the presence of receptor antagonists. Cortisol induced DNA damage and the production of RNS was further attenuated in the presence of an iNOS inhibitor. An increase in the expression of iNOS in response to psychological stress was observed in vivo and in cortisol-treated cells. Inhibition of glucocorticoid receptor-associated Src kinase also produced a decrease in cortisol-induced RNS. CONCLUSION: These results demonstrate that glucocorticoids may interact with iNOS in a non-genomic manner to produce damaging levels of RNS, thus allowing an insight into the potential mechanisms by which psychological stress may impact breast cancer.


Assuntos
Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Dano ao DNA , Glucocorticoides/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Espécies Reativas de Nitrogênio/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais , Animais , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Dano ao DNA/efeitos dos fármacos , Reparo do DNA , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glucocorticoides/farmacologia , Humanos , Camundongos , Modelos Biológicos , Óxido Nítrico Sintase Tipo II/genética , Estresse Oxidativo , Transdução de Sinais/efeitos dos fármacos
7.
PLoS Genet ; 6(9): e1001087, 2010 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-20824130

RESUMO

Abeta peptide accumulation is thought to be the primary event in the pathogenesis of Alzheimer's disease (AD), with downstream neurotoxic effects including the hyperphosphorylation of tau protein. Glycogen synthase kinase-3 (GSK-3) is increasingly implicated as playing a pivotal role in this amyloid cascade. We have developed an adult-onset Drosophila model of AD, using an inducible gene expression system to express Arctic mutant Abeta42 specifically in adult neurons, to avoid developmental effects. Abeta42 accumulated with age in these flies and they displayed increased mortality together with progressive neuronal dysfunction, but in the apparent absence of neuronal loss. This fly model can thus be used to examine the role of events during adulthood and early AD aetiology. Expression of Abeta42 in adult neurons increased GSK-3 activity, and inhibition of GSK-3 (either genetically or pharmacologically by lithium treatment) rescued Abeta42 toxicity. Abeta42 pathogenesis was also reduced by removal of endogenous fly tau; but, within the limits of detection of available methods, tau phosphorylation did not appear to be altered in flies expressing Abeta42. The GSK-3-mediated effects on Abeta42 toxicity appear to be at least in part mediated by tau-independent mechanisms, because the protective effect of lithium alone was greater than that of the removal of tau alone. Finally, Abeta42 levels were reduced upon GSK-3 inhibition, pointing to a direct role of GSK-3 in the regulation of Abeta42 peptide level, in the absence of APP processing. Our study points to the need both to identify the mechanisms by which GSK-3 modulates Abeta42 levels in the fly and to determine if similar mechanisms are present in mammals, and it supports the potential therapeutic use of GSK-3 inhibitors in AD.


Assuntos
Envelhecimento/patologia , Doença de Alzheimer/enzimologia , Doença de Alzheimer/patologia , Precursor de Proteína beta-Amiloide/metabolismo , Proteínas de Drosophila/antagonistas & inibidores , Drosophila melanogaster/enzimologia , Quinase 3 da Glicogênio Sintase/antagonistas & inibidores , Envelhecimento/efeitos dos fármacos , Doença de Alzheimer/mortalidade , Doença de Alzheimer/fisiopatologia , Precursor de Proteína beta-Amiloide/toxicidade , Animais , Modelos Animais de Doenças , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/efeitos dos fármacos , Genes Dominantes/genética , Quinase 3 da Glicogênio Sintase/metabolismo , Humanos , Lítio/farmacologia , Proteínas Mutantes/toxicidade , Sistema Nervoso/efeitos dos fármacos , Sistema Nervoso/metabolismo , Sistema Nervoso/patologia , Neurônios/efeitos dos fármacos , Neurônios/enzimologia , Neurônios/patologia , Peptídeos/toxicidade , Fosforilação/efeitos dos fármacos , Fosfosserina/metabolismo , Proteínas tau/metabolismo
8.
Gene ; 851: 147022, 2023 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-36347335

RESUMO

The response to psychological stress can differ depending on the type and duration of the stressor. Acute stress can facilitate a "fight or flight response" and aid survival, whereas chronic long-term stress with the persistent release of stress hormones such as cortisol has been shown to be detrimental to health. We are now beginning to understand how this stress hormone response impacts important processes such as DNA repair and cell proliferation processes in breast cancer. However, it is not known what epigenetic changes stress hormones induce in breast cancer. Epigenetic mechanisms include modification of DNA and histones within chromatin that may be involved in governing the transcriptional processes in cancer cells in response to changes by endogenous stress hormones. The contribution of endogenous acute or long-term exposure of glucocorticoid stress hormones, and exogenous glucocorticoids to methylation patterns in breast cancer tissues with different aetiologies remains to be evaluated. In vitro and in vivo models were developed to investigate the epigenetic modifications and their contribution to breast cancer progression and aetiology. A panel of triple negative breast cancer cell lines were treated with the glucocorticoid, cortisol which resulted in epigenetic alteration characterised by loss of methylation on promoter regions of tumour suppressor genes including ESR1, and loss of methylation on LINE-1 repetitive element used as a surrogate marker for global methylation. This was verified in vivo in MDA-MB-231 xenografts; the model verified the loss of methylation on ESR1 promoter, and subsequent increase in ESR1 expression in primary tumours in mice subjected to restraint stress. Our study highlights that DNA methylation landscape in breast cancer can be altered in response to stress and glucocorticoid treatment.


Assuntos
Receptor alfa de Estrogênio , Neoplasias de Mama Triplo Negativas , Humanos , Camundongos , Animais , Fulvestranto , Receptor alfa de Estrogênio/genética , Receptor alfa de Estrogênio/metabolismo , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Neoplasias de Mama Triplo Negativas/genética , Glucocorticoides/farmacologia , Hidrocortisona/farmacologia , Metilação de DNA
9.
J Biomech ; 152: 111528, 2023 05.
Artigo em Inglês | MEDLINE | ID: mdl-36989970

RESUMO

Following cervical spine fusion there is a reduction in maximum range of motion (ROM) but how this impacts activity of daily living (ADLs) and quality of life is unknown. This study's purpose is to quantify maximum and functional cervical spine ROM in patients with multi-level cervical fusion (>3 levels) compared to controls during ADLs and to correlate functional range of motion with scores from patient reported outcomes measures (PROs) including the Comparative Pain Scale (CPS), Fear Avoidance Belief Questionnaire (FABQ), and Neck Disability Index (NDI). An inertial measurement unit (IMU) system quantified ROM during ADLs in the extension/flexion, lateral bending, and axial rotation directions of motion. The reliability of this system was compared to standard optical motion tracking. Fourteen participants (8 females, age = 60.0 years (18.7) (median, (interquartile range)) with a history of multi-level cervical fusion (years post-op 0.9 (0.7)) were compared to 16 controls (13 females, age = 52.1 years (15.8)). PROs were collected for each participant. Fusion participants had significantly decreased maximum ROM in all directions of motion. Fusion participants had decreased ROM for some ADLs (backing up a car, using a phone, donning socks, negotiating stairs). CPS, FABQ, and NDI scores were significantly increased in fusion participants. Reductions in two activities (backing up a car, stair negotiation) correlated with a combination of increased PRO scores. Cervical fusion decreases maximum ROM and is correlated with increased PROs in some ADLs, however there is minimal impact on functional ROM. Investigation into velocity and acceleration may yield categorization of pathologic movement.


Assuntos
Atividades Cotidianas , Fusão Vertebral , Feminino , Humanos , Pessoa de Meia-Idade , Qualidade de Vida , Reprodutibilidade dos Testes , Vértebras Cervicais , Amplitude de Movimento Articular , Rotação , Fenômenos Biomecânicos
10.
J Pediatr Orthop ; 32(6): e35-9, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22892633

RESUMO

BACKGROUND: Current literature supporting treatment strategies for osteochondritis dissecans (OCD) has limited prognostic utility. Presently, there is no gold standard for an OCD "healing" outcome. Accurate and reliable determination of the OCD healed status on the basis of radiographs would be valuable to provide a guide for evidence-based treatment of OCD. The purpose of this study was to determine interrater and intrarater reliability of radiographic assessment of OCD lesion healing in the knee. METHODS: A hospital database identified 39 consecutive patients with OCD in 42 knees, who were treated conservatively for at least 6 months. Patients were an average age of 11.9 years (±2.4 y). A total of 102 presentation slides were prepared, each containing 2 similar radiographic views from the same patient taken 6 months (±1 mo) apart. These slides were then categorized by 4 reviewers (1 orthopaedic surgeon, 1 orthopaedic fellow, and 2 musculoskeletal radiologists) blinded to patient data, as healed, not healed, or unable to evaluate the OCD lesion. Reviewers repeated their assessment at a minimum of 3 weeks after their first read. Intrarater and interrater reliability were measured with the Cohen κ coefficient and Randolph's free-marginal multirater κ, respectively, and with the percent agreement. RESULTS: OCD lesion categorization demonstrated insufficient interrater reliability with a κ of 0.44 and 63% agreement. The notch view had the highest interrater reliability with a κ of 0.63% and 76% agreement, and the lateral view had the lowest interrater reliability with a κ of 0.29% and 53% agreement. The OCD lesion categorization demonstrated substantial intrarater reliability with a κ of 0.68% and 81% agreement. CONCLUSIONS: Reviewers did not consistently agree on the "healing" status of the OCD lesions on the basis of radiographic assessments. Standard criteria to assess healing are needed to consistently evaluate OCD knee lesions in children. LEVEL OF EVIDENCE: Not applicable.


Assuntos
Articulação do Joelho/diagnóstico por imagem , Osteocondrite Dissecante/diagnóstico por imagem , Adolescente , Criança , Bases de Dados Factuais , Medicina Baseada em Evidências , Humanos , Articulação do Joelho/patologia , Variações Dependentes do Observador , Osteocondrite Dissecante/patologia , Osteocondrite Dissecante/terapia , Radiografia , Reprodutibilidade dos Testes
11.
BMJ Open Sci ; 5(1): e100203, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-35047705

RESUMO

OBJECTIVE: The roll-out of the Pfizer-BioNTech BNT162b2 COVID-19 vaccine has brought many logistical challenges, such as the absence of comprehensive stability data leading to strict handling instructions during dilution and administration. Accidental mishandling therefore presents challenging clinical dilemmas, which often led vaccine providers to err on the side of caution and discard mishandled vials rather than risk administering ineffective vaccine. This study aims to answer key questions about the vaccine's stability to allow for a more informed decision-making process should a non-conformity occur. METHODS: Residual vaccine in freshly used, but appropriately stored vials collected from vaccination centres in Brighton, UK, were tested after exposure to various handling conditions and analysed by dynamic light scattering to determine the size of the lipid-mRNA nanoparticles, and gel electrophoresis to visualise the mRNA integrity and separation from the lipid formulation. RESULTS: Knocking or dropping vaccine samples from small heights resulted in lowest levels of instability, indicating low risk of compromising clinical efficacy. However, repeated drawing and injecting through 23 G needles at high speed and, more significantly, shaking and vortexing led to progressive increase in the size and polydispersity index of the lipid-mRNA nanoparticles, coupled with or caused by up to ~50% release of mRNA from the lipid formulation. This is thought to impact the vaccine's efficacy due to lack of free mRNA protection and cellular internalisation. CONCLUSIONS: These results reiterate the importance of adhering to the manufacturer's instructions on handling, especially with regard to shaking and exposing the vaccine to excessive vibration.

12.
BMC Genet ; 10: 77, 2009 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-19948010

RESUMO

BACKGROUND: The Glued gene of Drosophila melanogaster encodes the homologue of the vertebrate p150Glued subunit of dynactin. The Glued1 mutation compromises the dynein-dynactin retrograde motor complex and causes disruptions to the adult eye and the CNS, including sensory neurons and the formation of the giant fiber system neural circuit. RESULTS: We performed a 2-stage genetic screen to identify mutations that modified phenotypes caused by over-expression of a dominant-negative Glued protein. We screened over 34,000 flies and isolated 41 mutations that enhanced or suppressed an eye phenotype. Of these, 12 were assayed for interactions in the giant fiber system by which they altered a giant fiber morphological phenotype and/or altered synaptic function between the giant fiber and the tergotrochanteral muscle motorneuron. Six showed interactions including a new allele of atypical protein kinase C (aPKC). We show that this cell polarity regulator interacts with Glued during central synapse formation. We have mapped the five other interacting mutations to discrete chromosomal regions. CONCLUSION: Our results show that an efficient way to screen for genes involved in central synapse formation is to use a two-step strategy in which a screen for altered eye morphology precedes the analysis of central synaptogenesis. This has highlighted a role for aPKC in the formation of an identified central synapse.


Assuntos
Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Olho/metabolismo , Proteínas Associadas aos Microtúbulos/genética , Proteína Quinase C/genética , Sinapses/metabolismo , Animais , Polaridade Celular , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/citologia , Drosophila melanogaster/metabolismo , Olho/citologia , Feminino , Regulação da Expressão Gênica , Testes Genéticos , Masculino , Proteínas Associadas aos Microtúbulos/metabolismo , Mutação , Fenótipo , Proteína Quinase C/metabolismo , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , Sinapses/genética
13.
Cryobiology ; 58(1): 37-44, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-18983838

RESUMO

The drug efflux pump P-glycoprotein (P-gp) (ABCB1) confers multidrug resistance, a major cause of failure in the chemotherapy of tumours, exacerbated by a shortage of potent and selective inhibitors. A high throughput assay using purified P-gp to screen and characterise potential inhibitors would greatly accelerate their development. However, long-term stability of purified reconstituted ABCB1 can only be reliably achieved with storage at -80 degrees C. For example, at 20 degrees C, the activity of ABCB1 was abrogated with a half-life of <1 day. The aim of this investigation was to stabilise purified, reconstituted ABCB1 to enable storage at higher temperatures and thereby enable design of a high throughput assay system. The ABCB1 purification procedure was optimised to allow successful freeze drying by substitution of glycerol with the disaccharides trehalose or maltose. Addition of disaccharides resulted in ATPase activity being retained immediately following lyophilisation with no significant difference between the two disaccharides. However, during storage trehalose preserved ATPase activity for several months regardless of the temperature (e.g. 60% retention at 150 days), whereas ATPase activity in maltose purified P-gp was affected by both storage time and temperature. The data provide an effective mechanism for the production of resilient purified, reconstituted ABCB1.


Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/isolamento & purificação , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Dissacarídeos , Liofilização/métodos , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/química , Adenosina Trifosfatases/metabolismo , Animais , Varredura Diferencial de Calorimetria , Glicerol , Humanos , Maltose , Mariposas/genética , Transição de Fase , Proteolipídeos/química , Proteolipídeos/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Temperatura , Trealose
14.
Biosystems ; 184: 103996, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31369834

RESUMO

In this paper we present a mathematical solution that allows the elimination rate-constant or half life of a drug to be estimated from a single blood drug measurement. This is of great utility in clinical areas involving care of criticallly ill or vulnerable patients, where providing more than one blood sample can involve significant risks. The calculations used in our approach, based solely on a single sample, do not require complex pharmacokinetic software, but instead can be simply performed at the patient's bedside using standard personal computing tools. The proposed method allows a personalised estimate of the drug's half life, which is preferable to using population averages, or using estimates based on proxy markers of lagging organ function, which are both indirect and generally inaccurate for a patient with confounding factors.


Assuntos
Estado Terminal/terapia , Taxa de Depuração Metabólica , Preparações Farmacêuticas/metabolismo , Farmacocinética , Algoritmos , Ciclosporina/administração & dosagem , Ciclosporina/sangue , Ciclosporina/farmacocinética , Monitoramento de Medicamentos/métodos , Meia-Vida , Humanos , Imunossupressores/administração & dosagem , Imunossupressores/sangue , Imunossupressores/farmacocinética , Modelos Teóricos , Preparações Farmacêuticas/administração & dosagem , Software
15.
J Spec Pediatr Nurs ; 24(2): e12240, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30896893

RESUMO

PURPOSE: Current evidence-based research suggests that early evaluation, comprehensive care plans, and appropriate referrals for childhood and adolescent behavioral and development needs is critical for successful family-centered outcomes. The overall purpose of this study was to conduct an assessment of a state public health program that offers diagnostic evaluation and coordination for children with behavioral and developmental disorders in the state of Virginia (Child Development Center programs, or CDC). A secondary purpose was to provide translational policy and advocacy targets based on key findings. DESIGN AND METHOD: The evaluation of the scope of services of the CDC programs was done using qualitative interviews with a focus group interview (n = 23), interviews from representatives from individual centers ( n = 5 centers), and descriptive quantitative data elements for the fiscal year 2015. RESULTS: After conducting the state public health evaluation, several translational health policy priorities emerged, including: (a) the need for integrated data standards, (b) Lack of developmental pediatric workforce, particularly in rural sectors of the state, and (c) Need for enhanced program support for care coordination. CONCLUSION: Academic nurse and public health partnerships can aid in translation from research to policy among vulnerable populations and assist in communication to key stakeholders and legislators for iterative action and reassessment.


Assuntos
Transtornos do Comportamento Infantil/terapia , Serviços de Saúde da Criança/organização & administração , Deficiências do Desenvolvimento/terapia , Crianças com Deficiência/estatística & dados numéricos , Adolescente , Criança , Transtornos do Comportamento Infantil/prevenção & controle , Pré-Escolar , Deficiências do Desenvolvimento/prevenção & controle , Política de Saúde , Humanos , Avaliação das Necessidades , Qualidade da Assistência à Saúde , Fatores Socioeconômicos , Virginia
16.
Spine Deform ; 7(3): 404-409, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-31053310

RESUMO

STUDY DESIGN: In vitro biomechanical analysis. OBJECTIVES: Compare the destabilizing effects of anterior discectomy to posterior spinal releases. SUMMARY OF BACKGROUND DATA: Posterior release and pedicle screw fixation has become the accepted form of treatment for lumbar and thoracolumbar pediatric scoliotic spinal deformity. A biomechanical evaluation of posterior releases with comparison to traditional anterior releases has not been reported in the lumbar spine. METHODS: Eleven fresh-frozen human thoracolumbar specimens (T9-L5) were tested by a robotic manipulator (Staubli RX90; moment target of 5.0 Nm, force target of 50 N) in axial rotation (AR), plus lateral and anterior translation (LT and AT). Specimens underwent either sequential anterior release (partial and full discectomy) or posterior release (inferior facetectomy and wide posterior release) from T10 to L4. Partial discectomy retained the posterior 50% of disc and posterior longitudinal ligament, whereas full discectomy removed all of the disc and PLL. Wide posterior release included total facetectomy plus ligamentum flavum and spinous process resection. RESULTS: Inferior facetectomy produced an average increase of 1.5° ± 1.0° (p = .0625), 1.0 ± 0.8 mm (p = .0313), and 0.2 ± 0.3 mm (p = .156) in AR, LT, and AT, respectively. Compared with partial facetectomy, wide posterior release produced an average additional increase of 8.1° ± 4.0° (p = .0312), 2.0 ± 2.2 mm (p = .4062), and 1.1 ± 1.0 mm (p = .0625) in AR, LT, and AT, respectively. Full discectomy produced 201%, 161%, and 153% of the motion relative to wide posterior release in AR, LT, and AT, respectively (p = .0043, .0087, and .0173). Partial discectomy and wide posterior release proved statistically equivalent. CONCLUSIONS: Wide posterior release of the thoracolumbar spine allows significant correction and may be superior to inferior facetectomy in axial rotation. Although complete discectomy with PLL resection would likely allow greater correction, a more clinically realistic partial discectomy confers similar corrective potential in vitro compared with wide posterior release. LEVEL OF EVIDENCE: Not applicable.


Assuntos
Discotomia/métodos , Vértebras Lombares , Amplitude de Movimento Articular/fisiologia , Vértebras Torácicas , Idoso , Fenômenos Biomecânicos , Feminino , Humanos , Vértebras Lombares/fisiologia , Vértebras Lombares/cirurgia , Masculino , Pessoa de Meia-Idade , Procedimentos Ortopédicos/métodos , Vértebras Torácicas/fisiologia , Vértebras Torácicas/cirurgia
17.
Cancer Lett ; 459: 59-71, 2019 09 10.
Artigo em Inglês | MEDLINE | ID: mdl-31132432

RESUMO

Stress hormones have been shown to be important mediators in driving malignant growth and reducing treatment efficacy in breast cancer. Glucocorticoids can induce DNA damage through an inducible nitric oxide synthase (iNOS) mediated pathway to increase levels of nitric oxide (NO). Using an immune competent mouse breast cancer model and 66CL4 breast cancer cells we identified a novel role of NOS inhibition to reduce stress-induced breast cancer metastasis. On a mechanistic level we show that the glucocorticoid cortisol induces expression of keys genes associated with angiogenesis, as well as pro-tumourigenic immunomodulation. Transcriptomics analysis confirmed that in the lungs of tumour-bearing mice, stress significantly enriched pathways associated with tumourigenesis, some of which could be regulated with NOS inhibition. These results demonstrate the detrimental involvement of NOS in stress hormone signalling, and the potential future benefits of NOS inhibition in highly stressed patients.


Assuntos
Neoplasias da Mama/patologia , Inibidores Enzimáticos/farmacologia , Hidrocortisona/farmacologia , Neoplasias Mamárias Experimentais/patologia , Óxido Nítrico Sintase/antagonistas & inibidores , Estresse Psicológico/metabolismo , Animais , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/enzimologia , Neoplasias da Mama/genética , Linhagem Celular Tumoral , Dano ao DNA , Interações Medicamentosas , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/secundário , Células MCF-7 , Neoplasias Mamárias Experimentais/tratamento farmacológico , Neoplasias Mamárias Experimentais/enzimologia , Neoplasias Mamárias Experimentais/genética , Camundongos , Camundongos Endogâmicos BALB C , Mifepristona/farmacologia , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico/biossíntese , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase/metabolismo , Espécies Reativas de Nitrogênio/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Estresse Psicológico/patologia
18.
Methods Mol Biol ; 491: 177-86, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18998093

RESUMO

A number of recent studies have described the activation of BK(Ca) channels by steroid hormones such as estrogen. The proposed mechanisms are diverse and include both the direct interaction with the ion channel subunits and the stimulation via receptor activation and cell signalling pathways. To investigate the activation of BK(Ca) channels by estrogen we devised a cell-free system by incorporating recombinant channels of known subunit composition into artificial bilayers and recorded single channel currents. This chapter describes the methods used to prepare purified membrane fractions from cultured cells and the construction of artificial phospholipids bilayers for the incorporation and recording of ion channels.


Assuntos
Estrogênios/farmacologia , Subunidades alfa do Canal de Potássio Ativado por Cálcio de Condutância Alta/fisiologia , Bicamadas Lipídicas , Linhagem Celular , Eletrofisiologia/métodos , Humanos , Rim/embriologia , Subunidades alfa do Canal de Potássio Ativado por Cálcio de Condutância Alta/efeitos dos fármacos , Bicamadas Lipídicas/síntese química , Fosfatidiletanolaminas , Fosfatidilserinas , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologia
19.
Int J Pharm Pract ; 26(2): 174-182, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28349576

RESUMO

OBJECTIVE: There is substantial evidence demonstrating that transferring patients between care providers is a high-risk area for medicines management. This study aimed to investigate the effect of sending patients' hospital discharge letters to their nominated community pharmacists on the number of discrepancies between the patient's general practitioner (GP) records and the discharge letter and between the patient's self-described medication regime and the discharge letter. METHODS: In a randomised, controlled trial, 33 participants in two groups, control and intervention, had their discharge letter sent to either their GP only or their GP and nominated community pharmacy after hospital discharge. At least 3 weeks after hospital discharge, the participant's current GP's medication record and their self-described medication regime was obtained. Discrepancies between their GP medication record and their discharge letter and between the participant's self-described medication regime and their discharge letter were counted. The number of discrepancies (relative to the number of drugs prescribed) in the intervention group was compared with the control group for each of the above two categories, using the chi-squared test to determine the statistical significance of any differences between the two groups. RESULTS: The intervention group had statistically fewer discrepancies than the control group for both data sets: GP records compared with the discharge letters (P < 0.0005); participants' self-described medication regimes compared with the discharge letters (P < 0.00005). CONCLUSIONS: Sending a copy of patients' discharge letters to their community pharmacists could be beneficial in reducing post-discharge prescribing discrepancies and improving patient understanding of the changes made to their medicines.


Assuntos
Erros de Medicação/prevenção & controle , Reconciliação de Medicamentos/métodos , Sumários de Alta do Paciente Hospitalar , Transferência da Responsabilidade pelo Paciente/organização & administração , Farmacêuticos/organização & administração , Idoso , Feminino , Clínicos Gerais/organização & administração , Hospitais , Humanos , Masculino , Reconciliação de Medicamentos/organização & administração , Alta do Paciente
20.
Health Equity ; 2(1): 1-4, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30283844

RESUMO

Purpose: To examine geospatial gaps in identification and evaluation of children with special healthcare needs (CSHCN) within public child development centers (CDCs). Methods: A descriptive geospatial design was used to visually depict service gaps, proximity, and clustering of area-level sociodemographic attributes of Virginia counties, and patient-level data within each CDC. Results: Geospatial analysis shows population density of uninsured children against CDC resources. Data visualization facilitates policy advocacy based on the identification of care and screening gaps for CSHCN. Conclusion: This project illustrates the collaborative potential between researchers and Health Department members to identify gaps in access to care.

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