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1.
Nat Methods ; 16(8): 703-706, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31363206

RESUMO

Proteins can be phosphorylated at neighboring sites resulting in different functional states, and studying the regulation of these sites has been challenging. Here we present Thesaurus, a search engine that detects and quantifies phosphopeptide positional isomers from parallel reaction monitoring and data-independent acquisition mass spectrometry experiments. We apply Thesaurus to analyze phosphorylation events in the PI3K/AKT signaling pathway and show neighboring sites with distinct regulation.


Assuntos
Fosfopeptídeos/análise , Fosfoproteínas/análise , Proteoma/análise , Ferramenta de Busca/métodos , Células HeLa , Humanos , Proteínas Substratos do Receptor de Insulina/metabolismo , Isomerismo , Células MCF-7 , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais , Espectrometria de Massas em Tandem
2.
Mol Cell Proteomics ; 19(7): 1104-1119, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32234964

RESUMO

Stimulating brown adipose tissue (BAT) activity represents a promising therapy for overcoming metabolic diseases. mTORC2 is important for regulating BAT metabolism, but its downstream targets have not been fully characterized. In this study, we apply proteomics and phosphoproteomics to investigate the downstream effectors of mTORC2 in brown adipocytes. We compare wild-type controls to isogenic cells with an induced knockout of the mTORC2 subunit RICTOR (Rictor-iKO) by stimulating each with insulin for a 30-min time course. In Rictor-iKO cells, we identify decreases to the abundance of glycolytic and de novo lipogenesis enzymes, and increases to mitochondrial proteins as well as a set of proteins known to increase upon interferon stimulation. We also observe significant differences to basal phosphorylation because of chronic RICTOR loss including decreased phosphorylation of the lipid droplet protein perilipin-1 in Rictor-iKO cells, suggesting that RICTOR could be involved with regulating basal lipolysis or droplet dynamics. Finally, we observe mild dampening of acute insulin signaling response in Rictor-iKO cells, and a subset of AKT substrates exhibiting statistically significant dependence on RICTOR.


Assuntos
Adipócitos Marrons/efeitos dos fármacos , Adipócitos Marrons/metabolismo , Insulina/farmacocinética , Alvo Mecanístico do Complexo 2 de Rapamicina/metabolismo , Proteoma/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteína Companheira de mTOR Insensível à Rapamicina/metabolismo , Animais , Cromatografia Líquida , Técnicas de Inativação de Genes , Ontologia Genética , Glicólise/efeitos dos fármacos , Insulina/metabolismo , Lipogênese/efeitos dos fármacos , Camundongos , Mitocôndrias/efeitos dos fármacos , Fosforilação , Proteína Companheira de mTOR Insensível à Rapamicina/genética , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Espectrometria de Massas em Tandem
3.
J Community Health ; 46(4): 808-816, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-33389475

RESUMO

Serogroup B meningococcal disease (MenB) causes almost 60% of meningitis cases among adolescents and young adults. Yet, MenB vaccine coverage among adolescents remains below 10%. Since parents are the primary medical decision makers for adolescents, we examined MenB vaccination rates and parent attitudes about meningitis and the MenB vaccine. In 2018, in conjunction with a county-wide, school-based immunization campaign, we conducted a mixed methods study among parents of 16- to 17-year-olds. We facilitated focus groups asking parents about their knowledge of meningitis and reactions to educational materials and sent behavioral surveys based on Health Belief Model constructs to parents through the county high school system. Parents in three focus groups (n = 8; participation rate = 13%) expressed confusion about their child's need to receive the MenB vaccine in addition to the meningococcal conjugate vaccine (MenACWY), but conveyed strong trust in their physicians' recommendation. Among survey participants (n = 170), 70 (41%) had heard of the MenB vaccine. Among those 70 parents, the most common barriers to vaccination were concerns about side effects (55%) and uncertainty of susceptibility due to receipt of the MenACWY vaccine (30%). The percentage of teens that received at least one dose of the MenB vaccine was 50% (n = 35) by parent report and 23% (n = 16) by state vaccination records. Parents demonstrated uncertainty and confusion about the MenB vaccine particularly due to the existence of another meningitis vaccine and limited health care provider recommendations. Confirmatory studies of parent confusion about the MenB vaccine are needed to develop interventions.


Assuntos
Conhecimentos, Atitudes e Prática em Saúde , Infecções Meningocócicas , Vacinas Meningocócicas , Neisseria meningitidis Sorogrupo B , Pais , Adolescente , Humanos , Infecções Meningocócicas/prevenção & controle , Instituições Acadêmicas , Estudantes , Vacinação , Adulto Jovem
4.
J Occup Environ Hyg ; 18(7): 305-313, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34038318

RESUMO

Recently, total inward leakage (TIL) for filtering facepiece and elastomeric half-mask respirators (EHRs) was measured according to the International Organization for Standardization (ISO) test method standard 16900-1:2014 that showed larger TIL for corn oil aerosol than for NaCl aerosol. Comparison of TIL measured for different aerosols for higher protection level respirators is lacking. The objective of this study was to determine TIL for EHRs, full-facepiece respirators, and loose-fitting and tight-fitting powered air-purifying respirators (PAPRs) using NaCl and corn oil aerosols to compare. TIL was measured for two models each of EHRs, full-facepiece respirators, and loose-fitting and tight-fitting PAPRs. After fit testing with a PortaCount (TSI, St. Paul, MN) using the Occupational Safety and Health Administration (OSHA) protocol, eight subjects were tested in the NaCl aerosol chamber first and then in the corn oil aerosol chamber, while another eight subjects tested in the reverse order. Subjects were randomly assigned to one of the two groups. TIL was measured as a ratio of mass-based aerosol concentrations inside the mask to the test chamber while the subjects performed ISO 16900-1-defined exercises using continuous sampling methods. The concentration of corn oil aerosol was measured with one light scattering photometer, alternately, and NaCl aerosol was measured using two flame photometers. Results showed the geometric mean TIL for EHR was significantly (p < 0.05) larger for corn oil aerosol than for NaCl aerosol. EHR models equipped with P100 filters showed relatively smaller TIL values than the same models with N95 filters showing that TIL was inversely related to filter efficiency. Interestingly, TIL was significantly (p < 0.05) larger for NaCl aerosol than for corn oil aerosol for PAPRs, but not for full-facepiece respirators. TIL was inversely related to fit factors of respirator types. Overall, filter efficiency and faceseal leakage determine TIL. The relative trends in TIL for the two aerosols' test methods differ between respirator types indicating that generalization of TIL for respirator types may not be appropriate when using different test agents.


Assuntos
Exposição Ocupacional , Dispositivos de Proteção Respiratória , Aerossóis , Óleo de Milho , Filtração , Humanos , National Institute for Occupational Safety and Health, U.S. , Exposição Ocupacional/prevenção & controle , Tamanho da Partícula , Cloreto de Sódio/análise , Estados Unidos
5.
Artigo em Inglês | MEDLINE | ID: mdl-33720803

RESUMO

Measurement of skin exposure to particles using interception (e.g., cotton gloves) and removal (e.g., wiping) sampling techniques could be inaccurate because these substrates do not have the same topography and adhesion characteristics as skin. The objective of this study was to compare particle transfer and adherence to cotton gloves, cotton gloves with artificial sebum, and a pre-moistened polyvinyl alcohol (PVA) material with bare human skin (fingertip, palm). Experiments were performed with aluminum oxide powder under standardized conditions for three types of surfaces touched, applied loads, contact times, and powder mass levels. In the final mixed model, the fixed effects of substrate, surface type, applied load, and powder mass and their significant two-way interaction terms explained 71% (transfer) and 74% (adherence) of the observed total variance in measurements. For particle mass transfer, compared with bare skin, bias was -77% (cotton glove with sebum) to +197% (PVA material) and for adherence bias ranged from -40% (cotton glove) to +428% (PVA material), which indicated under- and over-sampling by these substrates, respectively. Dermal exposure assessment would benefit from sampling substrates that better reflect human skin characteristics and more accurately estimate exposures. Mischaracterization of dermal exposure has important implications for exposure and risk assessment.


Assuntos
Exposição Ambiental/análise , Pele/metabolismo , Manejo de Espécimes , Adesividade , Óxido de Alumínio/análise , Óxido de Alumínio/química , Óxido de Alumínio/metabolismo , Fibra de Algodão , Humanos , Álcool de Polivinil/química , Pós/análise , Pós/química , Pós/metabolismo , Absorção Cutânea
6.
Angew Chem Int Ed Engl ; 60(20): 11487-11493, 2021 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-33683786

RESUMO

Electrochemical carbon dioxide (CO2 ) reduction reaction (CO2 RR) is an attractive approach to deal with the emission of CO2 and to produce valuable fuels and chemicals in a carbon-neutral way. Many efforts have been devoted to boost the activity and selectivity of high-value multicarbon products (C2+ ) on Cu-based electrocatalysts. However, Cu-based CO2 RR electrocatalysts suffer from poor catalytic stability mainly due to the structural degradation and loss of active species under CO2 RR condition. To date, most reported Cu-based electrocatalysts present stabilities over dozens of hours, which limits the advance of Cu-based electrocatalysts for CO2 RR. Herein, a porous chlorine-doped Cu electrocatalyst exhibits high C2+ Faradaic efficiency (FE) of 53.8 % at -1.00 V versus reversible hydrogen electrode (VRHE ). Importantly, the catalyst exhibited an outstanding catalytic stability in long-term electrocatalysis over 240 h. Experimental results show that the chlorine-induced stable cationic Cu0 /Cu+ species and the well-preserved structure with abundant active sites are critical to the high FE of C2+ in the long-term run of electrochemical CO2 reduction.

7.
MMWR Morb Mortal Wkly Rep ; 69(19): 587-590, 2020 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-32407300

RESUMO

An estimated 2.1 million U.S. adults are housed within approximately 5,000 correctional and detention facilities† on any given day (1). Many facilities face significant challenges in controlling the spread of highly infectious pathogens such as SARS-CoV-2, the virus that causes coronavirus disease 2019 (COVID-19). Such challenges include crowded dormitories, shared lavatories, limited medical and isolation resources, daily entry and exit of staff members and visitors, continual introduction of newly incarcerated or detained persons, and transport of incarcerated or detained persons in multiperson vehicles for court-related, medical, or security reasons (2,3). During April 22-28, 2020, aggregate data on COVID-19 cases were reported to CDC by 37 of 54 state and territorial health department jurisdictions. Thirty-two (86%) jurisdictions reported at least one laboratory-confirmed case from a total of 420 correctional and detention facilities. Among these facilities, COVID-19 was diagnosed in 4,893 incarcerated or detained persons and 2,778 facility staff members, resulting in 88 deaths in incarcerated or detained persons and 15 deaths among staff members. Prompt identification of COVID-19 cases and consistent application of prevention measures, such as symptom screening and quarantine, are critical to protecting incarcerated and detained persons and staff members.


Assuntos
Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/epidemiologia , Pneumonia Viral/diagnóstico , Pneumonia Viral/epidemiologia , Prisões , COVID-19 , Infecções por Coronavirus/mortalidade , Infecções por Coronavirus/prevenção & controle , Humanos , Pandemias/prevenção & controle , Pneumonia Viral/mortalidade , Pneumonia Viral/prevenção & controle , Prevalência , SARS-CoV-2 , Estados Unidos/epidemiologia
8.
J Intensive Care Med ; 35(8): 805-809, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30089431

RESUMO

BACKGROUND: There is no consensus for the length of prophylactic antibiotics after delayed chest closure (DCC) postcardiac surgery in pediatrics. In September 2014, our institution's pediatric cardiac intensive care unit changed the policy on length of prophylactic antibiotics after DCC from 5 days (control) to 2 days (study group). The objective of the study was to determine whether a 2-day course of antibiotics is as effective as a 5-day course in preventing blood stream and sternal wound infections in pediatric DCC. METHODS: Retrospective and prospective study. Primary end points included incidence of sternal wound infections and positive sternal imaging for infection. Surrogate markers of infection were collected at 4 time points. RESULTS: During the study period, 139 patients had DCC postcardiac surgery of which 110 patients were included for analysis, 54 patients in the control and 56 in the study group. There was no difference in total number of positive wound cultures/chest computed tomography (CT) findings (4/54 [7.5%] control vs 5/56 [8.9%] study group, P = .3), positive blood cultures (P = .586), median postsurgical length of stay (P = .4), or readmissions within 30 days postsurgery (P = .6). All secondary end points were similar in both groups except peak heart rate between weeks 2 and 4 (P = .041). CONCLUSION: Two days of prophylactic antibiotics is not inferior to 5 days of prophylactic antibiotics after DCC following pediatric cardiac surgery.


Assuntos
Antibioticoprofilaxia/métodos , Procedimentos Cirúrgicos Cardíacos/efeitos adversos , Cuidados Pós-Operatórios/métodos , Sepse/prevenção & controle , Infecção da Ferida Cirúrgica/prevenção & controle , Antibioticoprofilaxia/normas , Biomarcadores/sangue , Criança , Pré-Escolar , Esquema de Medicação , Feminino , Humanos , Lactente , Masculino , Pediatria/métodos , Pediatria/normas , Cuidados Pós-Operatórios/normas , Estudos Prospectivos , Estudos Retrospectivos , Esterno/microbiologia , Esterno/cirurgia , Fatores de Tempo , Tomografia Computadorizada por Raios X
9.
J Clin Psychol ; 76(1): 137-145, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31468533

RESUMO

OBJECTIVE: Neuropsychiatric symptoms are a major component of dementia irrespective of severity or subtype. We aimed to determine the feasibility of biographical films to reduce neuropsychiatric symptoms in people with moderate to severe dementia over a 32-week period. METHOD: A total of 11 people with dementia situated in a residential care home took part in this mixed-method feasibility study. Carers reported neuropsychiatric symptoms of residents at three time-points, and their experience of the study was obtained at a feedback session. RESULTS: There was a significant reduction in neuropsychiatric symptoms in residents with neuropsychiatric impairment from baseline to the end of study (p = .042; d = .98). Thematic analysis identified three major themes: Triggered memories, knowledge gained to support care, and perceived changes in the resident. CONCLUSION: The findings suggest that it is feasible to use biographical films long-term to reduce neuropsychiatric symptoms of dementia, alongside routine care.


Assuntos
Sintomas Comportamentais/terapia , Demência/complicações , Filmes Cinematográficos , Psicoterapia/métodos , Idoso , Sintomas Comportamentais/etiologia , Estudos de Viabilidade , Feminino , Humanos , Masculino , Memória Episódica , Casas de Saúde
10.
Nat Methods ; 13(5): 431-4, 2016 05.
Artigo em Inglês | MEDLINE | ID: mdl-27018578

RESUMO

Systematic approaches to studying cellular signaling require phosphoproteomic techniques that reproducibly measure the same phosphopeptides across multiple replicates, conditions, and time points. Here we present a method to mine information from large-scale, heterogeneous phosphoproteomics data sets to rapidly generate robust targeted mass spectrometry (MS) assays. We demonstrate the performance of our method by interrogating the IGF-1/AKT signaling pathway, showing that even rarely observed phosphorylation events can be consistently detected and precisely quantified.


Assuntos
Fator de Crescimento Insulin-Like I/metabolismo , Fosfopeptídeos/metabolismo , Proteômica/métodos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Biologia de Sistemas/métodos , Técnicas de Cultura de Células , Mineração de Dados , Bases de Dados Genéticas , Ensaios de Triagem em Larga Escala , Humanos , Fator de Crescimento Insulin-Like I/farmacologia , Células MCF-7 , Fosforilação , Transdução de Sinais , Espectrometria de Massas em Tandem
11.
J Occup Environ Hyg ; 16(7): 489-497, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31107187

RESUMO

An ASTM International subcommittee on Respiratory Protection, F23.65 is currently developing a consensus standard for assessing respirator fit capability (RFC) criteria of half-facepiece air-purifying particulate respirators. The objective of this study was to evaluate if the test methods being developed for half-facepiece respirators can reasonably be applied to nonpowered full-facepiece-air-purifying respirators (FF-APR). Benchmark RFC test data were collected for three families of FF-APRs (a one-size-only family, a two-size family, and a three-size family). All respirators were equipped with P100 class particulate filters. Respirators were outfitted with a sampling probe to collect an in-mask particle concentration sample in the breathing zone of the wearer. Each of the six respirator facepieces was tested on the National Institute for Occupational Safety and Health 25-subject Bivariate Panel. The RFC test assessed face seal leakage using a PortaCount fit test. Subjects followed the corresponding Occupational Safety and Health Administration-accepted fit test protocol. Two donnings per subject/respirator model combination were performed. The panel passing rate (PPR) (number or percentage of subjects in the panel achieving acceptable fit on at least one of two donnings) was determined for each respirator family at specified fit factor passing levels of 500, 1,000, and 2,000. As a reasonable expectation based on a previous analysis of alpha and beta fit test errors for various panel sizes, the selected PPR benchmark for our study was >75%. At the fit factor passing level of 500 obtained on at least one of two donnings, the PPRs for three-, two-, and one-size families were 100, 79, and 88%, respectively. As the fit factor passing criterion increased from 500 to 1,000 or 2,000, PPRs followed a decreasing trend. Each of the three tested families of FF-APRs are capable of fitting ≥75% of the intended user population at the 500 fit factor passing level obtained on at least one of two donnings. The methods presented here can be used as a reference for standards development organizations considering developing RFC test requirements.


Assuntos
Máscaras/normas , Dispositivos de Proteção Respiratória/normas , Adulto , Humanos , Masculino , Teste de Materiais/métodos , Pessoa de Meia-Idade , National Institute for Occupational Safety and Health, U.S. , Exposição Ocupacional/prevenção & controle , Estados Unidos
12.
J Biol Chem ; 292(50): 20481-20493, 2017 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-28982975

RESUMO

Lipins 1, 2, and 3 are Mg2+-dependent phosphatidic acid phosphatases and catalyze the penultimate step of triacylglycerol synthesis. We have previously investigated the biochemistry of lipins 1 and 2 and shown that di-anionic phosphatidic acid (PA) augments their activity and lipid binding and that lipin 1 activity is negatively regulated by phosphorylation. In the present study, we show that phosphorylation does not affect the catalytic activity of lipin 3 or its ability to associate with PA in vitro The lipin proteins each contain a conserved polybasic domain (PBD) composed of nine lysine and arginine residues located between the conserved N- and C-terminal domains. In lipin 1, the PBD is the site of PA binding and sensing of the PA electrostatic charge. The specific arrangement and number of the lysines and arginines of the PBD vary among the lipins. We show that the different PBDs of lipins 1 and 3 are responsible for the presence of phosphoregulation on the former but not the latter enzyme. To do so, we generated lipin 1 that contained the PBD of lipin 3 and vice versa. The lipin 1 enzyme with the lipin 3 PBD lost its ability to be regulated by phosphorylation but remained downstream of phosphorylation by mammalian target of rapamycin. Conversely, the presence of the lipin 1 PBD in lipin 3 subjected the enzyme to negative intramolecular control by phosphorylation. These results indicate a mechanism for the observed differences in lipin phosphoregulation in vitro.


Assuntos
Proteínas Nucleares/metabolismo , Fosfatidato Fosfatase/metabolismo , Ácidos Fosfatídicos/metabolismo , Processamento de Proteína Pós-Traducional , Células 3T3-L1 , Sequência de Aminoácidos , Substituição de Aminoácidos , Animais , Sítios de Ligação , Sequência Conservada , Células HeLa , Humanos , Cinética , Lipossomos , Camundongos , Micelas , Mutação , Proteínas Nucleares/química , Proteínas Nucleares/genética , Fosfatidato Fosfatase/química , Fosfatidato Fosfatase/genética , Fosforilação , Domínios e Motivos de Interação entre Proteínas , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/metabolismo
13.
J Occup Environ Hyg ; 15(8): 616-627, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29781773

RESUMO

The International Organization for Standardization (ISO) standard 16900-1:2014 specifies the use of sodium chloride (NaCl) and corn oil aerosols, and sulfur hexafluoride gas for measuring total inward leakage (TIL). However, a comparison of TIL between different agents is lacking. The objective of this study was to measure and compare TIL for respirators using corn oil and NaCl aerosols. TIL was measured with 10 subjects donning two models of filtering facepiece respirators (FFRs) including FFP1, N95, P100, and elastomeric half-mask respirators (ERs) in NaCl and corn oil aerosol test chambers, using continuous sampling methods. After fit testing with a PortaCount (TSI, Inc., St. Paul, MN) using the Occupational Safety and Health Administration (OSHA) protocol, five subjects were tested in the NaCl chamber first and then in the corn oil chamber, while other subjects tested in the reverse order. TIL was measured as a ratio of mass-based aerosol concentrations in-mask to the test chamber, while the subjects performed ISO 16900-1-defined exercises. The concentration of NaCl aerosol was measured using two flame photometers, and corn oil aerosol was measured with one light scattering photometer. The same instruments were used to measure filter penetration in both chambers using a Plexiglas setup. The size distribution of aerosols was determined using a scanning mobility particle sizer and charge was measured with an electrometer. Filter efficiency was measured using an 8130 Automated Filter Tester (TSI). Results showed the geometric mean TIL for corn oil aerosol for one model each of all respirator categories, except P100, were significantly (p < 0.05) greater than for NaCl aerosol. Filter penetration in the two test chambers showed a trend similar to TIL. The count median diameter was ∼82 nm for NaCl and ∼200 nm for corn oil aerosols. The net positive charge for NaCl aerosol was relatively larger. Both fit factor and filter efficiency influence TIL measurement. Overall, TIL determination with aerosols of different size distributions and charges using different methodologies may produce dissimilar results.


Assuntos
Óleo de Milho/análise , Dispositivos de Proteção Respiratória/normas , Cloreto de Sódio/análise , Aerossóis/análise , Face/anatomia & histologia , Filtração/instrumentação , Humanos , Exposição por Inalação , Tamanho da Partícula , Fotometria
14.
Biochim Biophys Acta ; 1858(8): 1868-75, 2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-27155567

RESUMO

The Escherichia coli sensor kinase EnvZ modulates porin expression in response to various stimuli, including extracellular osmolarity, the presence of procaine and interaction with an accessory protein, MzrA. Two major outer membrane porins, OmpF and OmpC, act as passive diffusion-limited pores that allow compounds, including certain classes of antibiotics such as ß-lactams and fluoroquinolones, to enter the bacterial cell. Even though the mechanisms by which EnvZ detects and processes the presence of various stimuli are a fundamental component of microbial physiology, they are not yet fully understood. Here, we assess the role of TM1 during signal transduction in response to the presence of extracellular osmolarity. Various mechanisms of transmembrane communication have been proposed including rotation of individual helices within the transmembrane domain, dynamic movement of the membrane-distal portion of the cytoplasmic domain and regulated intra-protein unfolding. To assess these possibilities, we have created a library of single-Cys-containing EnvZ proteins in order to facilitate sulfhydryl-reactivity experimentation. Our results demonstrate that the major TM1-TM1' interface falls along a single surface consisting of residue positions 19, 23, 26, 30 and 34. In addition, we show that Cys substitutions within the N- and C-terminal regions of TM1 result in drastic changes to EnvZ signal output. Finally, we demonstrate that core residues within TM1 are responsible for both TM1 dimerisation and maintenance of steady-state signal output. Overall, our results suggest that no major rearrangement of the TM1-TM1' interface occurs during transmembrane communication in response to extracellular osmolarity. We conclude by discussing these results within the frameworks of several proposed models for transmembrane communication.


Assuntos
Proteínas da Membrana Bacteriana Externa/química , Proteínas de Escherichia coli/química , Complexos Multienzimáticos/química , Regulação Alostérica , Substituição de Aminoácidos , Proteínas da Membrana Bacteriana Externa/genética , Cisteína/química , Dimerização , Proteínas de Escherichia coli/genética , Complexos Multienzimáticos/genética , Mutagênese Sítio-Dirigida , Concentração Osmolar , Domínios Proteicos , Estrutura Secundária de Proteína , Proteínas Recombinantes/química , Transdução de Sinais/fisiologia
15.
Mol Syst Biol ; 12(12): 888, 2016 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-27909043

RESUMO

The coordinated regulation of protein kinases is a rapid mechanism that integrates diverse cues and swiftly determines appropriate cellular responses. However, our understanding of cellular decision-making has been limited by the small number of simultaneously monitored phospho-regulatory events. Here, we have estimated changes in activity in 215 human kinases in 399 conditions derived from a large compilation of phosphopeptide quantifications. This atlas identifies commonly regulated kinases as those that are central in the signaling network and defines the logic relationships between kinase pairs. Co-regulation along the conditions predicts kinase-complex and kinase-substrate associations. Additionally, the kinase regulation profile acts as a molecular fingerprint to identify related and opposing signaling states. Using this atlas, we identified essential mediators of stem cell differentiation, modulators of Salmonella infection, and new targets of AKT1. This provides a global view of human phosphorylation-based signaling and the necessary context to better understand kinase-driven decision-making.


Assuntos
Células-Tronco Embrionárias Humanas/citologia , Proteínas Quinases/metabolismo , Biologia de Sistemas/métodos , Diferenciação Celular , Bases de Dados de Proteínas , Regulação Enzimológica da Expressão Gênica , Células HeLa , Células-Tronco Embrionárias Humanas/enzimologia , Humanos , Fosforilação , Proteômica/métodos , Transdução de Sinais
16.
Inorg Chem ; 56(24): 15304-15313, 2017 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-29210577

RESUMO

The phosphorescent emitters are essential to realize energy-efficient display and lighting panels. The solution processability is of particular interest for large-scale and low-cost production. Here, we present a series of the heteroleptic iridium (Ir) complexes, Ir(ppy)2L1, Ir(ppy)2L2, and Ir(ppy)2L3, using the new ancillary ligands, including 1-(2-chlorophenyl)-5-hydroxy-3-methyl-1H-pyrazole-4-carbaldehyde (L1), 5-hydroxy-3-methyl-1-(p-tolyl)-1H-pyrazole-4-carbaldehyde (L2), and 5-hydroxy-3-methyl-1-phenyl-1H-pyrazole-4-carbaldehyde (L3). Their photophysical and electrochemical properties were systematically characterized, followed by comparing with those predicted by density functional theory simulations using hybrid functionals. Among the three phosphors synthesized, Ir(ppy)2L1 exhibits the highest photoluminescence quantum yield (ΦPL = 89%), with an exciton lifetime of 0.34 µs. By using 4,4'-bis(carbazole-9-yl)biphenyl as the host material, we demonstrate high current efficiencies of 64 and 40 cd A-1 at 100 cd m-2 in its vacuum-evaporated and solution-processed organic light-emitting devices, respectively, revealing the promise for large-area light sources.

17.
J Occup Environ Hyg ; 14(7): 540-550, 2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28440728

RESUMO

Printing devices are known to emit chemicals into the indoor atmosphere. Understanding factors that influence release of chemical contaminants from printers is necessary to develop effective exposure assessment and control strategies. In this study, a desktop fused deposition modeling (FDM) 3-dimensional (3-D) printer using acrylonitrile butadiene styrene (ABS) or polylactic acid (PLA) filaments and two monochrome laser printers were evaluated in a 0.5 m3 chamber. During printing, chamber air was monitored for vapors using a real-time photoionization detector (results expressed as isobutylene equivalents) to measure total volatile organic compound (TVOC) concentrations, evacuated canisters to identify specific VOCs by off-line gas chromatography-mass spectrometry (GC-MS) analysis, and liquid bubblers to identify carbonyl compounds by GC-MS. Airborne particles were collected on filters for off-line analysis using scanning electron microscopy with an energy dispersive x-ray detector to identify elemental constituents. For 3-D printing, TVOC emission rates were influenced by a printer malfunction, filament type, and to a lesser extent, by filament color; however, rates were not influenced by the number of printer nozzles used or the manufacturer's provided cover. TVOC emission rates were significantly lower for the 3-D printer (49-3552 µg h-1) compared to the laser printers (5782-7735 µg h-1). A total of 14 VOCs were identified during 3-D printing that were not present during laser printing. 3-D printed objects continued to off-gas styrene, indicating potential for continued exposure after the print job is completed. Carbonyl reaction products were likely formed from emissions of the 3-D printer, including 4-oxopentanal. Ultrafine particles generated by the 3-D printer using ABS and a laser printer contained chromium. Consideration of the factors that influenced the release of chemical contaminants (including known and suspected asthmagens such as styrene and 4-oxopentanal) from a FDM 3-D printer should be made when designing exposure assessment and control strategies.


Assuntos
Poluição do Ar em Ambientes Fechados/análise , Material Particulado/análise , Impressão Tridimensional , Compostos Orgânicos Voláteis/análise , Acrilonitrila/análise , Aldeídos/análise , Butadienos , Cromo/análise , Monitoramento Ambiental/métodos , Cetonas/análise , Poliésteres , Estireno/análise
18.
Clin Infect Dis ; 63(7): 878-888, 2016 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-27358357

RESUMO

BACKGROUND: During 2009 and 2010, 2 clusters of organ transplant-transmitted Balamuthia mandrillaris, a free-living ameba, were detected by recognition of severe unexpected illness in multiple recipients from the same donor. METHODS: We investigated all recipients and the 2 donors through interview, medical record review, and testing of available specimens retrospectively. Surviving recipients were tested and treated prospectively. RESULTS: In the 2009 cluster of illness, 2 kidney recipients were infected and 1 died. The donor had Balamuthia encephalitis confirmed on autopsy. In the 2010 cluster, the liver and kidney-pancreas recipients developed Balamuthia encephalitis and died. The donor had a clinical syndrome consistent with Balamuthia infection and serologic evidence of infection. In both clusters, the 2 asymptomatic recipients were treated expectantly and survived; 1 asymptomatic recipient in each cluster had serologic evidence of exposure that decreased over time. Both donors had been presumptively diagnosed with other neurologic diseases prior to organ procurement. CONCLUSIONS: Balamuthia can be transmitted through organ transplantation with an observed incubation time of 17-24 days. Clinicians should be aware of Balamuthia as a cause of encephalitis with high rate of fatality, and should notify public health departments and evaluate transplant recipients from donors with signs of possible encephalitis to facilitate early diagnosis and targeted treatment. Organ procurement organizations and transplant centers should be aware of the potential for Balamuthia infection in donors with possible encephalitis and also assess donors carefully for signs of neurologic infection that may have been misdiagnosed as stroke or as noninfectious forms of encephalitis.


Assuntos
Amebíase , Balamuthia mandrillaris , Encefalite , Transplante de Rim/efeitos adversos , Transplante de Fígado/efeitos adversos , Adulto , Amebíase/diagnóstico por imagem , Amebíase/patologia , Amebíase/transmissão , Encéfalo/diagnóstico por imagem , Encéfalo/parasitologia , Encéfalo/patologia , Criança , Pré-Escolar , Encefalite/diagnóstico por imagem , Encefalite/patologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Doadores de Tecidos , Transplantados
19.
Prev Med ; 89: 237-244, 2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-27311334

RESUMO

Places where we buy food influence dietary patterns, making local food environments a good example of a mass influence on population diets. Cross-cultural studies, using reliable methods, may help understanding the relationship between food environments and diet-related health outcomes. We aimed to understand cross-national differences in the local food environment between Madrid and Baltimore by comparing an average neighborhood in each city in terms of food store types, healthy food availability, and residents' pedestrian access. During 2012-2013, we assessed one neighborhood (~15,000 residents) in each city selecting median areas in terms of socio-demographic characteristics (segregation, education, aging, and population density). We collected on-field data on (a) number and types of all food stores, (b) overall healthy food availability and (c) specific availability of fruits & vegetables. Throughout a street network analysis (200m, 400m and 800m) of food stores with high healthy food availability, we estimated residents' pedestrian accessibility. We found 40 stores in Madrid and 14 in Baltimore. Small food stores carrying fresh foods in Madrid contrasted with the high presence of corner and chain convenience stores in Baltimore. In Madrid, 77% of the residents lived within less than 200m from a food store with high healthy food availability. In contrast, 95% of Baltimore's residents lived further than 400m from these stores. Our results may help promoting interventions from local city agencies to allocate resources to existing small-sized food stores, and to improve walkable urban environments. These actions may influence food choices, especially for those residents lacking access to private vehicles.


Assuntos
Comparação Transcultural , Meio Ambiente , Abastecimento de Alimentos , Estudos de Casos Organizacionais , Características de Residência , Baltimore , Comércio , Dieta , Frutas , Promoção da Saúde/métodos , Humanos , Densidade Demográfica , Pobreza , Espanha , População Urbana , Verduras
20.
Analyst ; 141(6): 1997-2008, 2016 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-26878279

RESUMO

Biotechnology, separation science, and clinical research are impacted by microfluidic devices. Separation and manipulation of bioparticles such as DNA, protein and viruses are performed on these platforms. Microfluidic systems provide many attractive features, including small sample size, rapid detection, high sensitivity and short processing time. Dielectrophoresis (DEP) and electrophoresis are especially well suited to microscale bioparticle control and have been demonstrated in many formats. In this work, an optimized gradient insulator-based DEP device was utilized for concentration of Sindbis virus, an animal virus with a diameter of 68 nm. Within only a few seconds, the concentration of Sindbis virus can be increased by two to six times in the channel under easily accessible voltages as low as about 70 V. Compared with traditional diagnostic methods used in virology, DEP-based microfluidics can enable faster isolation, detection and concentration of viruses in a single step within a short time.


Assuntos
Eletroforese/instrumentação , Sindbis virus/isolamento & purificação , Análise de Elementos Finitos , Dispositivos Lab-On-A-Chip
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