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2.
Opt Express ; 28(26): 38773-38787, 2020 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-33379439

RESUMO

Early detection of a gas kick is crucial for preventing uncontrolled blowout that could cause loss of life, loss of assets, and environmental damage. Multiphase flow experiments conducted in this research demonstrate the capability of downhole fiber optic sensors to detect a potential gas influx in real-time in a 5000 ft deep wellbore. Gas rise velocities estimated independently using fiber optic distributed acoustic sensor (DAS), distributed temperature sensor (DTS), downhole gauges, surface measurements, and multiphase flow correlations show good agreement in each case, demonstrating reliability in the assessment. Real-time data visualization was implemented on a secure cloud-based platform to improve computational efficiency. This study provides novel insights on the effect of circulation rates, gas kick volumes, backpressure, and injection methods on gas rise dynamics in a full-scale wellbore.

3.
Plants (Basel) ; 9(2)2020 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-32085566

RESUMO

Plant genomes provide information on biosynthetic pathways involved in the production of industrially relevant compounds. Genome size estimates are essential for the initiation of genome projects. The genome size of rooibos (Aspalathus linearis species complex) was estimated using DAPI flow cytometry and k-mer analyses. For flow cytometry, a suitable nuclei isolation buffer, plant tissue and a transport medium for rooibos ecotype samples collected from distant locations were identified. When using radicles from commercial rooibos seedlings, Woody Plant Buffer and Vicia faba as an internal standard, the flow cytometry-estimated genome size of rooibos was 1.24 ± 0.01 Gbp. The estimates for eight wild rooibos growth types did not deviate significantly from this value. K-mer analysis was performed using Illumina paired-end sequencing data from one commercial rooibos genotype. For biocomputational estimation of the genome size, four k-mer analysis methods were investigated: A standard formula and three popular programs (BBNorm, GenomeScope, and FindGSE). GenomeScope estimates were strongly affected by parameter settings, specifically CovMax. When using the complete k-mer frequency histogram (up to 9 × 105), the programs did not deviate significantly, estimating an average rooibos genome size of 1.03 ± 0.04 Gbp. Differences between the flow cytometry and biocomputational estimates are discussed.

4.
Sensors (Basel) ; 20(1)2020 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-31906506

RESUMO

Effective well control depends on the drilling teams' knowledge of wellbore flow dynamics and their ability to predict and control influx. Unfortunately, detection of a gas influx in an offshore environment is particularly challenging, and there are no existing datasets that have been verified and validated for gas kick migration at full-scale annular conditions. This study bridges this gap and presents pioneering research in the application of fiber optic sensing for monitoring gas in riser. The proposed sensing paradigm was validated through well-scale experiments conducted at Petroleum Engineering Research & Technology Transfer lab (PERTT) facility at Louisiana State University (LSU), simulating an offshore marine riser environment with its larger than average annular space and mud circulation capability. The experimental setup instrumented with distributed fiber optic sensors and pressure/temperature gauges provides a physical model to study the dynamic gas migration in full-scale annular conditions. Current kick detection methods primarily utilize surface measurements and do not always reliably detect a gas influx. The proposed application of distributed fiber optic sensing overcomes this key limitation of conventional kick detection methods, by providing real-time distributed downhole data for accurate and reliable monitoring. The two-phase flow experiments conducted in this research provide critical insights for understanding the flow dynamics in offshore drilling riser conditions, and the results provide an indication of how quickly gas can migrate in a marine riser scenario, warranting further investigation for the sake of effective well control.

5.
BioTech (Basel) ; 9(4)2020 Sep 23.
Artigo em Inglês | MEDLINE | ID: mdl-35822822

RESUMO

Rooibos (Aspalathus linearis), widely known as a herbal tea, is endemic to the Cape Floristic Region of South Africa (SA). It produces a wide range of phenolic compounds that have been associated with diverse health promoting properties of the plant. The species comprises several growth forms that differ in their morphology and biochemical composition, only one of which is cultivated and used commercially. Here, we established methodologies for non-invasive transcriptome research of wild-growing South African plant species, including (1) harvesting and transport of plant material suitable for RNA sequencing; (2) inexpensive, high-throughput biochemical sample screening; (3) extraction of high-quality RNA from recalcitrant, polysaccharide- and polyphenol rich plant material; and (4) biocomputational analysis of Illumina sequencing data, together with the evaluation of programs for transcriptome assembly (Trinity, IDBA-Trans, SOAPdenovo-Trans, CLC), protein prediction, as well as functional and taxonomic transcript annotation. In the process, we established a biochemically characterized sample pool from 44 distinct rooibos ecotypes (1-5 harvests) and generated four in-depth annotated transcriptomes (each comprising on average ≈86,000 transcripts) from rooibos plants that represent distinct growth forms and differ in their biochemical profiles. These resources will serve future rooibos research and plant breeding endeavours.

6.
Ann Gen Psychiatry ; 18: 13, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31413721

RESUMO

BACKGROUND: Pharmacogenomics is starting to build momentum in clinical utility, perhaps the most in mental and behavioral healthcare. However, efficient delivery of this information to the point of prescribing remains a significant challenge. Clinical decision support has an opportunity to address this void by integrating pharmacogenomics into the clinician workflow. METHODS: To address the specific needs of mental health clinicians at the point of care, we conducted 3 focus groups with a total of 16 mental health clinicians. Each 1-h focus group was designed to identify the desired clinical decision support features, with a particular interest in pharmacogenomics, and potential negative or unintended consequences of clinical decision support integration at the point of care in a mental healthcare setting. We implemented an iterative design to expand upon knowledge generated in prior focus groups. The results from the guided discussion in the first focus group were used to develop a mental health clinical decision support prototype. This prototype was then presented during the next two focus groups to drive the discussion. RESULTS: This study has identified main themes related to the desired clinical decision support features of mental health clinicians, the use of pharmacogenomics in practice, and unintended and negative consequences of clinical decision support integration at the point of care. Clinicians desire a more complete picture of the medication history of patients and guidance to choose medications in relation to cost, insurance coverage, and pharmacogenetics interactions. Mental health clinicians agreed that pharmacogenetics is useful and impacts their prescribing decisions when the data are available. Several negative consequences of clinical decision support integration were identified including alert fatigue and frustration using the tool. Several points of contention were related to the integration of the clinical decision support with the electronic health record, including bidirectional flow of information, speed, location within workflow, and potential incompleteness of information. CONCLUSIONS: We have identified general and unique considerations of mental health clinicians with regard to clinical decision support. Clinical decision support that incorporates desired features while avoiding negative and unintended consequences will increase clinician usage and will have the potential to improve the care of patients.

8.
Appl Microbiol Biotechnol ; 103(11): 4429-4441, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30972461

RESUMO

Biosurfactants are amphiphilic molecules that interact with the surfaces of liquids leading to many useful applications. Most biosurfactants have been identified from cultured microbial sources, leaving a largely untapped resource of uncultured bacteria with potentially novel biosurfactant structures. To access the uncultured bacteria, a metagenomic library was constructed in Escherichia coli from environmental DNA within an E. coli, Pseudomonas putida and Streptomyces lividans shuttle vector. Phenotypic screening of the library in E. coli and P. putida by the paraffin spray assay identified a P. putida clone with biosurfactant activity. Sequence analysis and transposon mutagenesis confirmed that an ornithine acyl-ACP N-acyltransferase was responsible for the activity. Although the fosmid was not active in E. coli, overexpression of the olsB gene could be achieved under the control of the inducible T7 promoter, resulting in lyso-ornithine lipid production and biosurfactant activity in the culture supernatants. Screening for activity in more than one host increases the range of sequences that can be identified through metagenomic, since olsB would not have been identified if only E. coli had been used as a host. The potential of lyso-ornithine lipids as a biosurfactant has not been fully explored. Here, we present several biosurfactant parameters of lyso-ornithine lipid to assess its suitability for industrial application.


Assuntos
Acetiltransferases/metabolismo , Metagenômica/métodos , Ornitina/análogos & derivados , Tensoativos/metabolismo , Acetiltransferases/genética , Elementos de DNA Transponíveis , Escherichia coli/genética , Escherichia coli/metabolismo , Biblioteca Gênica , Testes Genéticos , Vetores Genéticos , Lipídeos , Mutagênese Insercional , Ornitina/metabolismo , Pseudomonas putida/genética , Pseudomonas putida/metabolismo , Análise de Sequência de DNA
9.
Molecules ; 24(5)2019 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-30866512

RESUMO

Aspalathin, the main polyphenol of rooibos (Aspalathus linearis), is associated with diverse health promoting properties of the tea. During fermentation, aspalathin is oxidized and concentrations are significantly reduced. Standardized methods for quality control of rooibos products do not investigate aspalathin, since current techniques of aspalathin detection require expensive equipment and expertise. Here, we describe a simple and fast thin-layer chromatography (TLC) method that can reproducibly visualize aspalathin in rooibos herbal tea and plant extracts at a limit of detection (LOD) equal to 178.7 ng and a limit of quantification (LOQ) equal to 541.6 ng. Aspalathin is a rare compound, so far only found in A. linearis and its (rare) sister species A. pendula. Therefore, aspalathin could serve as a marker compound for authentication and quality control of rooibos products, and the described TLC method represents a cost-effective approach for high-throughput screening of plant and herbal tea extracts.


Assuntos
Aspalathus/química , Chalconas/análise , Chás de Ervas/normas , Cromatografia em Camada Fina , Ensaios de Triagem em Larga Escala , Extratos Vegetais/normas , Controle de Qualidade
10.
Obes Surg ; 27(6): 1501-1507, 2017 06.
Artigo em Inglês | MEDLINE | ID: mdl-27885537

RESUMO

BACKGROUND: Non-anesthesia administered propofol (NAAP) has been shown to be a safe and effective method of sedation for patients undergoing gastrointestinal endoscopy. Bariatric surgery patients are potentially at a higher risk for sedation-related complications due to co-morbidities including obstructive sleep apnea. The outcomes of NAAP in bariatric patients have not been previously reported. METHODS: In this retrospective cohort study, severely obese patients undergoing pre-surgical outpatient esophagogastroduodenoscopy (EGD) were compared to non-obese control patients (BMI ≤ 25 kg/m2) undergoing diagnostic EGD at our institution from March 2011-September 2015 using our endoscopy database. Patients' demographics and procedural and recovery data, including any airway interventions, were statistically analyzed. RESULTS: We included 130 consecutive pre-operative bariatric surgical patients with average BMI 45.8 kg/m2 (range 34-80) and 265 control patients with average BMI 21.9 kg/m2 (range 14-25). The severely obese group had a higher prevalence of sleep apnea (62 vs 8%; p < 0.001), experienced more oxygen desaturations (22 vs 7%; p < 0.001), and received more chin lift maneuvers (20 vs 6%; p < 0.001). Advanced airway interventions were rarely required in either group and were not more frequent in the bariatric group. CONCLUSIONS: With appropriate training of endoscopy personnel, NAAP is a safe method of sedation in severely obese patients undergoing outpatient upper endoscopy.


Assuntos
Sedação Consciente , Gastroscopia , Hipnóticos e Sedativos/administração & dosagem , Obesidade Mórbida/cirurgia , Propofol/administração & dosagem , Adulto , Idoso , Idoso de 80 Anos ou mais , Assistência Ambulatorial , Estudos de Coortes , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Segurança do Paciente , Complicações Pós-Operatórias , Prevalência , Estudos Retrospectivos , Utah
11.
J Med Econ ; 19(5): 469-76, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26671481

RESUMO

OBJECTIVES: This study evaluated the effect of paliperidone palmitate long-acting injectable (LAI) antipsychotic on recovery-oriented mental health outcomes from the perspective of healthcare providers and patients during the treatment of patients with schizophrenia or schizoaffective disorders. METHODS: Archival data for patients with a primary diagnosis of schizophrenia or schizoaffective disorder receiving ≥6 months of paliperidone palmitate LAI were retrieved from the electronic medical records system at the Mental Health Center of Denver. Mental health recovery was assessed from both a provider's (Recovery Markers Inventory [RMI]) and patient's (Consumer Recovery Measure [CRM]) perspective. A three-level hierarchical linear model (HLM) was utilized to determine changes in CRM and RMI scores by including independent variables in the models: intercept, months from treatment (slope), treatment time period (pretreatment and treatment), age, gender, primary diagnosis, substance abuse diagnosis, concurrent medications, and adherence to paliperidone palmitate LAI. RESULTS: A total of 219 patients were identified and included in the study. Results of the final three-level HLMs indicated an overall increase in CRM scores (p < 0.05), an overall increase (p < 0.01), and an increased rate of change (p < 0.05) in RMI scores during the paliperidone palmitate LAI treatment period vs the pre-treatment period. LIMITATIONS: This study contained a retrospective, non-comparative design, and did not adjust for multiplicity Conclusions: The current study demonstrates that changes in recovery-oriented mental health outcomes can be detected following the administration of a specific antipsychotic treatment in persons with schizophrenia or schizoaffective disorders. Furthermore, patients receiving paliperidone palmitate LAI can effectively improve recovery-oriented outcomes, thereby supporting the drug's use as schizophrenia treatment from a recovery-oriented perspective.


Assuntos
Antipsicóticos/uso terapêutico , Palmitato de Paliperidona/uso terapêutico , Transtornos Psicóticos/tratamento farmacológico , Esquizofrenia/tratamento farmacológico , Adulto , Antipsicóticos/administração & dosagem , Preparações de Ação Retardada , Etnicidade , Feminino , Hospitalização/economia , Hospitalização/estatística & dados numéricos , Humanos , Masculino , Pessoa de Meia-Idade , Palmitato de Paliperidona/administração & dosagem , Prisões/economia , Prisões/estatística & dados numéricos , Psicometria , Estudos Retrospectivos , Apoio Social , Centros de Tratamento de Abuso de Substâncias/economia , Centros de Tratamento de Abuso de Substâncias/estatística & dados numéricos , Fatores de Tempo , Resultado do Tratamento
12.
J Christ Nurs ; 32(4): 236-41, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26548177

RESUMO

Healthcare literature suggests that many nurses fail to address patients' spiritual needs and/or identify signs of spiritual distress. A study was conducted to explore whether nurses in a medical center possessed the knowledge to assess patients' spirituality and design and implement a plan of spiritual care. The Spiritual Care Competence Scale was used to assess competence in spiritual care assessment and implementation; professionalization and improving quality; personal support and patient counseling; referral; attitude toward patient spirituality; and communication of spiritual needs.


Assuntos
Cristianismo , Enfermagem Holística/métodos , Competência Profissional , Terapias Espirituais/instrumentação , Espiritualidade , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Relações Enfermeiro-Paciente , Sudeste dos Estados Unidos , Terapias Espirituais/métodos , Adulto Jovem
13.
PLoS One ; 10(8): e0130533, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26244639

RESUMO

BACKGROUND: Beta-defensins (hBDs) provide antimicrobial and chemotactic defense against bacterial, viral and fungal infections. Human ß-defensin-2 (hBD-2) acts against gram-negative bacteria and chemoattracts immature dendritic cells, thus regulating innate and adaptive immunity. Immunosuppression due to hyperglycemia underlies chronic infection in Type 2 diabetes. Hyperglycemia also elevates production of dicarbonyls methylgloxal (MGO) and glyoxal (GO). METHODS: The effect of dicarbonyl on defensin peptide structure was tested by exposing recombinant hBD-2 (rhBD-2) to MGO or GO with subsequent analysis by MALDI-TOF MS and LC/MS/MS. Antimicrobial function of untreated rhBD-2 vs. rhBD-2 exposed to dicarbonyl against strains of both gram-negative and gram-positive bacteria in culture was determined by radial diffusion assay. The effect of dicarbonyl on rhBD-2 chemotactic function was determined by chemotaxis assay in CEM-SS cells. RESULTS: MGO or GO in vitro irreversibly adducts to the rhBD-2 peptide, and significantly reduces antimicrobial and chemotactic functions. Adducts derive from two arginine residues, Arg22 and Arg23 near the C-terminus, and the N-terminal glycine (Gly1). We show by radial diffusion testing on gram-negative E. coli and P. aeruginosa, and gram-positive S. aureus, and a chemotaxis assay for CEM-SS cells, that antimicrobial activity and chemotactic function of rhBD-2 are significantly reduced by MGO. CONCLUSIONS: Dicarbonyl modification of cationic antimicrobial peptides represents a potential link between hyperglycemia and the clinical manifestation of increased susceptibility to infection, protracted wound healing, and chronic inflammation in undiagnosed and uncontrolled Type 2 diabetes.


Assuntos
Bactérias/imunologia , Infecções Bacterianas/imunologia , Glioxal/análogos & derivados , beta-Defensinas/química , beta-Defensinas/imunologia , Imunidade Adaptativa , Antibacterianos/química , Antibacterianos/imunologia , Glioxal/imunologia , Humanos , Imunidade Inata , Metilação , Proteínas Recombinantes/química , Proteínas Recombinantes/imunologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
14.
J Vis Exp ; (98)2015 Apr 14.
Artigo em Inglês | MEDLINE | ID: mdl-25938804

RESUMO

Fluorescent proteins, fluorescent dyes and fluorophores in general have revolutionized the field of molecular cell biology. In particular, the discovery of fluorescent proteins and their genes have enabled the engineering of protein fusions for localization, the analysis of transcriptional activation and translation of proteins of interest, or the general tracking of individual cells and cell populations. The use of fluorescent protein genes in combination with retroviral technology has further allowed the expression of these proteins in mammalian cells in a stable and reliable manner. Shown here is how one can utilize these genes to give cells within a population of cells their own biosignature. As the biosignature is achieved with retroviral technology, cells are barcoded 'indefinitely'. As such, they can be individually tracked within a mixture of barcoded cells and utilized in more complex biological applications. The tracking of distinct populations in a mixture of cells is ideal for multiplexed applications such as discovery of drugs against a multitude of targets or the activation profile of different promoters. The protocol describes how to elegantly develop and amplify barcoded mammalian cells with distinct genetic fluorescent markers, and how to use several markers at once or one marker at different intensities. Finally, the protocol describes how the cells can be further utilized in combination with cell-based assays to increase the power of analysis through multiplexing.


Assuntos
Código de Barras de DNA Taxonômico/métodos , Proteínas Luminescentes/genética , Engenharia de Proteínas/métodos , Animais , Corantes Fluorescentes/química , Marcadores Genéticos/genética , Proteínas de Fluorescência Verde/química , Proteínas de Fluorescência Verde/genética , Humanos , Proteínas Luminescentes/química , Regiões Promotoras Genéticas , Retroviridae/genética
15.
J Biomol Screen ; 20(5): 616-26, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25724189

RESUMO

The DenV pre-membrane protein (prM) is a crucial chaperone for the viral envelope protein, preventing premature fusion with vesicles during viral export. prM molecules in immature particles are cleaved by host proteases, leading to mature fusogenic virions. Blockade of prM cleavage would restrict fusion and represents a novel druggable opportunity against DenV. We have thus established a cell-based platform to monitor prM processing that relies on an engineered two-tag scaffold that travels to the cell surface through the secretory pathway. The assay discriminates between a single cell-surface tag when prM is cleaved and two tags when it is not, as detected through fluorescent-coupled antibodies by flow cytometry. The assay, miniaturized into a 96-well plate format, was multiplexed with the HIV-1 envelope boundary, also cleaved in the same pathway. A pilot screen against 1280 compounds was executed, leading to the identification of a potential active and corroborating the robustness of our assay for large-scale screening. We describe for the first time a cell-based assay that monitors DenV prM processing within the classical secretory pathway, which was exploited to identify a potential novel drug against DenV.


Assuntos
Antivirais/farmacologia , Vírus da Dengue/efeitos dos fármacos , Descoberta de Drogas/métodos , Ensaios de Triagem em Larga Escala , Proteínas da Matriz Viral/antagonistas & inibidores , Linhagem Celular , Vírus da Dengue/metabolismo , Relação Dose-Resposta a Droga , Humanos , Concentração Inibidora 50 , Testes de Sensibilidade Microbiana , Bibliotecas de Moléculas Pequenas
16.
PLoS Pathog ; 10(4): e1004045, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24722773

RESUMO

Coxsackievirus B3 (CVB3), a member of the picornavirus family and enterovirus genus, causes viral myocarditis, aseptic meningitis, and pancreatitis in humans. We genetically engineered a unique molecular marker, "fluorescent timer" protein, within our infectious CVB3 clone and isolated a high-titer recombinant viral stock (Timer-CVB3) following transfection in HeLa cells. "Fluorescent timer" protein undergoes slow conversion of fluorescence from green to red over time, and Timer-CVB3 can be utilized to track virus infection and dissemination in real time. Upon infection with Timer-CVB3, HeLa cells, neural progenitor and stem cells (NPSCs), and C2C12 myoblast cells slowly changed fluorescence from green to red over 72 hours as determined by fluorescence microscopy or flow cytometric analysis. The conversion of "fluorescent timer" protein in HeLa cells infected with Timer-CVB3 could be interrupted by fixation, suggesting that the fluorophore was stabilized by formaldehyde cross-linking reactions. Induction of a type I interferon response or ribavirin treatment reduced the progression of cell-to-cell virus spread in HeLa cells or NPSCs infected with Timer-CVB3. Time lapse photography of partially differentiated NPSCs infected with Timer-CVB3 revealed substantial intracellular membrane remodeling and the assembly of discrete virus replication organelles which changed fluorescence color in an asynchronous fashion within the cell. "Fluorescent timer" protein colocalized closely with viral 3A protein within virus replication organelles. Intriguingly, infection of partially differentiated NPSCs or C2C12 myoblast cells induced the release of abundant extracellular microvesicles (EMVs) containing matured "fluorescent timer" protein and infectious virus representing a novel route of virus dissemination. CVB3 virions were readily observed within purified EMVs by transmission electron microscopy, and infectious virus was identified within low-density isopycnic iodixanol gradient fractions consistent with membrane association. The preferential detection of the lipidated form of LC3 protein (LC3 II) in released EMVs harboring infectious virus suggests that the autophagy pathway plays a crucial role in microvesicle shedding and virus release, similar to a process previously described as autophagosome-mediated exit without lysis (AWOL) observed during poliovirus replication. Through the use of this novel recombinant virus which provides more dynamic information from static fluorescent images, we hope to gain a better understanding of CVB3 tropism, intracellular membrane reorganization, and virus-associated microvesicle dissemination within the host.


Assuntos
Micropartículas Derivadas de Células/virologia , Enterovirus Humano B/fisiologia , Infecções por Enterovirus/metabolismo , Fagossomos/virologia , Eliminação de Partículas Virais/fisiologia , Animais , Micropartículas Derivadas de Células/genética , Micropartículas Derivadas de Células/metabolismo , Infecções por Enterovirus/genética , Células HeLa , Humanos , Camundongos , Proteínas Associadas aos Microtúbulos/genética , Proteínas Associadas aos Microtúbulos/metabolismo , Fagossomos/genética , Fagossomos/metabolismo , Proteínas Virais/genética , Proteínas Virais/metabolismo
17.
J Neuroinflammation ; 10: 127, 2013 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-24139179

RESUMO

BACKGROUND: The human ß-defensins (hBDs) are a highly conserved family of cationic antimicrobial and immunomodulatory peptides expressed primarily by epithelial cells in response to invasion by bacteria, fungi and some viruses. To date, the most studied members of this family of peptides are hBD-1, -2, and -3. Expression of hBD-1 and -2 has been demonstrated previously in cultured microglia and astrocytes of both mouse and human brain. Unlike inducible hBD-2 and -3, hBD-1 is constitutively expressed and is not generally upregulated by proinflammatory factors. In this study, we investigated whether hBDs, as active components of the innate immune response, are affected by pathological events in the Alzheimer's disease (AD) brain. We assessed the expression of hBD-1, -2, and -3 in tissue obtained at autopsy from AD and age-matched control brains. METHODS: Fixed and frozen choroid plexus and the CA1 region of the hippocampus were obtained at autopsy from individuals diagnosed with AD, or from age-matched control brains without diagnosed neurodegenerative disease. Histopathologically diagnosed AD brain tissue was obtained for our study. Immunocytochemical analysis was performed using affinity purified polyclonal antibodies directed against hBD-1, -2, or -3. TaqMan gene expression assays were used to quantify the mRNA of hBD-1, -2, and -3 in the choroid plexus and hippocampus. Immunocytochemical detection of iron deposits was achieved using a modified Perl's stain for redox-active iron. In vitro experiments were performed on human primary oral epithelial cells to model the human choroid plexus epithelial response to ferric chloride. Cells were then exposed to ferric chloride added to selected wells at 0, 1, or 10 mM concentrations for 24 h at 37°C. Total mRNA was isolated to quantify hBD-1 mRNA expression by RTqPCR. RESULTS: hBD-1 peptide is apparent in astrocytes of the AD hippocampus and hippocampal neurons, notably within granulovacuolar degeneration structures (GVD). A higher level of hBD-1 was also seen in the choroid plexus of AD brain in comparison to age-matched control tissue. Increased expression of hBD-1 mRNA was observed only in the choroid plexus of the AD brain when compared to expression level in age-matched control brain. Redox-active iron was also elevated in the AD choroid plexus and in vitro addition of Fe⁺³Cl3 to cultured epithelial cells induced hBD-1 mRNA expression. CONCLUSIONS: Our findings suggest interplay between hBD-1 and neuroimmunological responses in AD, marked by microglial and astrocytic activation, and increased expression of the peptide within the choroid plexus and accumulation within GVD. As a constitutively expressed component of the innate immune system, we propose that hBD-1 may be of considerable importance early in the disease process. We also demonstrate that increased iron deposition in AD may contribute to the elevated expression of hBD-1 within the choroid plexus. These findings represent a potentially important etiological aspect of Alzheimer's disease neuropathology not previously reported.


Assuntos
Doença de Alzheimer/metabolismo , Biomarcadores/análise , Plexo Corióideo/metabolismo , Hipocampo/metabolismo , beta-Defensinas/biossíntese , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Imuno-Histoquímica , Ferro/metabolismo , Masculino , Análise de Sequência com Séries de Oligonucleotídeos , Oxirredução , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Regulação para Cima
18.
ScientificWorldJournal ; 2012: 905785, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22606066

RESUMO

It is widely accepted that the brain responds to mechanical trauma and development of most neurodegenerative diseases with an inflammatory sequelae that was once thought exclusive to systemic immunity. Mostly cationic peptides, such as the ß-defensins, originally assigned an antimicrobial function are now recognized as mediators of both innate and adaptive immunity. Herein supporting evidence is presented for the hypothesis that neuropathological changes associated with chronic disease conditions of the CNS involve abnormal expression and regulatory function of specific antimicrobial peptides. It is also proposed that these alterations exacerbate proinflammatory conditions within the brain that ultimately potentiate the neurodegenerative process.


Assuntos
Peptídeos Catiônicos Antimicrobianos/imunologia , Doenças Neurodegenerativas/imunologia , beta-Defensinas/imunologia , Imunidade Adaptativa , Animais , Peptídeos Catiônicos Antimicrobianos/metabolismo , Apoptose , Astrócitos/imunologia , Astrócitos/metabolismo , Encéfalo/imunologia , Encéfalo/metabolismo , Encéfalo/patologia , Proliferação de Células , Doenças do Sistema Nervoso Central/imunologia , Doenças do Sistema Nervoso Central/metabolismo , Doenças do Sistema Nervoso Central/patologia , Quimiotaxia , Plexo Corióideo/imunologia , Plexo Corióideo/metabolismo , Citocinas/imunologia , Células Dendríticas/imunologia , Humanos , Imunidade Inata , Mediadores da Inflamação/imunologia , Microglia/imunologia , Microglia/metabolismo , Doenças Neurodegenerativas/metabolismo , Doenças Neurodegenerativas/patologia , Cicatrização , beta-Defensinas/metabolismo
19.
J Amino Acids ; 2011: 461216, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22332001

RESUMO

Neurodegeneration results from abnormalities in cerebral metabolism and energy balance within neurons, astrocytes, microglia, or microvascular endothelial cells of the blood-brain barrier. In Alzheimer's disease, ß-amyloid is considered the primary contributor to neuropathology and neurodegeneration. It now is believed that certain systemic diseases, such as diabetes mellitus, can contribute to neurodegeneration through the effects of chronic hyperglycemia/insulin resistance resulting in protein glycation, oxidative stress and inflammation within susceptible brain regions. Here, we present an overview of research focusing on the role of protein glycation, oxidative stress, and inflammation in the neurodegenerative process. Of special interest in this paper is the effect of methylglyoxal (MGO), a cytotoxic byproduct of glucose metabolism, elevated in neurodegenerative disease, and diabetes mellitus, on cerebral protein function and oxidative stress. How MGO interacts with amino acid residues within ß-amyloid, and small peptides within the brain, is also discussed in terms of the affect on protein function.

20.
Appl Opt ; 48(27): 5155-63, 2009 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-19767933

RESUMO

Grinding, lapping, and polishing are finishing processes used to achieve critical surface parameters in a variety of precision optical and electronic components. As these processes remove material from the surface through mechanical and chemical interactions, they may induce a damaged layer of cracks, voids, and stressed material below the surface. This subsurface damage (SSD) can degrade the performance of a final product by creating optical aberrations due to diffraction, premature failure in oscillating components, and a reduction in the laser induced damage threshold of high energy optics. As these defects lie beneath the surface, they are difficult to detect, and while many methods are available to detect SSD, they can have notable limitations regarding sample size and type, preparation time, or can be destructive in nature. The authors tested a nondestructive method for assessing SSD that consisted of tagging the abrasive slurries used in lapping and polishing with quantum dots (nano-sized fluorescent particles). Subsequent detection of fluorescence on the processed surface is hypothesized to indicate SSD. Quantum dots that were introduced to glass surfaces during the lapping process were retained through subsequent polishing and cleaning processes. The quantum dots were successfully imaged by both wide field and confocal fluorescence microscopy techniques. The detected fluorescence highlighted features that were not observable with optical or interferometric microscopy. Atomic force microscopy and additional confocal microscope analysis indicate that the dots are firmly embedded in the surface but do not appear to travel deep into fractures beneath the surface. Etching of the samples exhibiting fluorescence confirmed that SSD existed. SSD-free samples exposed to quantum dots did not retain the dots in their surfaces, even when polished in the presence of quantum dots.

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