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1.
ACS Chem Neurosci ; 11(5): 674-701, 2020 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-32065744

RESUMO

Competitive antagonists for ionotropic glutamate receptors (iGluRs) are highly valuable tool compounds for studying health and disease states in the central nervous system. However, only few subtype selective tool compounds are available and the discovery of antagonists with novel iGluR subtype selectivity profiles remains a profound challenge. In this paper, we report an elaborate structure-activity relationship (SAR) study of the parental scaffold 2,3-trans-3-carboxy-3-phenyl-proline by the synthesis of 40 new analogues. Three synthetic strategies were employed with two new strategies of which one being a highly efficient and fully enantioselective strategy based on C(sp3)-H activation methodology. The SAR study led to the conclusion that selectivity for the NMDA receptors was a general trend when adding substituents in the 5'-position. Selective NMDA receptor antagonists were obtained with high potency (IC50 values as low as 200 nM) and 3-34-fold preference for GluN1/GluN2A over GluN1/GluN2B-D NMDA receptors.

2.
Biomacromolecules ; 21(2): 356-365, 2020 02 10.
Artigo em Inglês | MEDLINE | ID: mdl-31860278

RESUMO

We present a method for reproducible manufacture of multiassay platforms with tunable mechanical properties for muscle tissue strip analysis. The platforms result from stereolithographic 3D printing of low protein-binding poly(ethylene glycol) diacrylate (PEGDA) hydrogels. Contractile microtissues have previously been engineered by immobilizing suspended cells in a confined hydrogel matrix with embedded anchoring cantilevers to facilitate muscle tissue strip formation. The 3D shape and mechanical properties of the confinement and the embedded cantilevers are critical for the tissue robustness. High-resolution 3D printing of PEGDA hydrogels offers full design freedom to engineer cantilever stiffness, while minimizing unwanted cell attachment. We demonstrate the applicability by generating suspended muscle tissue strips from C2C12 mouse myoblasts in a compliant fibrin-based hydrogel matrix. The full design freedom allows for new platform geometries that reduce local stress in the matrix and tissue, thus, reducing the risk of tissue fracture.

3.
Scand J Caring Sci ; 2019 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-31487064

RESUMO

Illness behaviour effects the quality of life of patients with chronic obstructive pulmonary disease (COPD) but is scarcely described in literature. The aim of this study was to explore self-reported illness behaviour of patients with COPD, who have declined nonpharmacological rehabilitation. The study has a qualitative design using semi-structured interviews. The subsequent analysis is a theory-guided mapping of actions reported by the patients in order to manage symptoms. These actions are understood and categorised according to the styles of coping described by Alonzo. Four categories of illness behaviour are identified: containment of symptoms, coping with symptoms through formal and informal interventions, adjustment of situations through compensating and economising interventions and crisis coping by surrendering. The analysis shows that behaviour that may seem unhelpful from a healthcare perspective may seem rational in the everyday life perspective of the patient. Findings show that reluctance to participation in rehabilitation should not only be interpreted as lack of motivation or health literacy. In the patients' perspective, nonpharmacological interventions might be perceived as a threat that could tip the delicate balance of everyday life with severe COPD.

4.
PLoS One ; 13(7): e0201380, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30052651

RESUMO

BACKGROUND: Non-gonococcal urethritis (NGU) is a common syndrome in men. NGU may have several causes, but many cases are caused by sexually transmitted infections that may also cause complications in their female partners. Chlamydia trachomatis and Mycoplasma genitalium are the most common causes of NGU, but in up to 35% of the cases, none of the known viral or bacterial causes are found. Traditionally, pathogens have been detected using various culture techniques that may not identify all species present in the urethra. To address this, we used culture-independent methods for analysis of the male urethral microbiota. METHODS: This case-control study analysed first void urine samples, collected at STD clinics in Stockholm, Sweden from men with idiopathic urethritis (IU), i.e. negative for Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium, Ureaplasma urealyticum, Trichomonas vaginalis, adenovirus, and herpes simplex virus type 1 and -2 together with samples from men without urethritis. Forty-six controls and 39 idiopathic urethritis patients were analysed. RESULTS: The microbiota was highly diverse: None of the 302 operational taxonomic units (OTUs) found in negative controls and IU patients were found in all of the samples or even in all of the samples in one group. More than 50% of the OTUs were only found in one or two of the total of 85 samples. Still the most dominant 1/6 of the genera constituted 79% of the sequences. Hierarchical clustering in a heatmap showed no specific clustering of patients or controls. A number of IU patient samples were dominated by a single genus previously related to urethritis (Gardnerella, Haemophilus, Ureaplasma). CONCLUSION: The male urethra contain a very diverse composition of bacteria, even in healthy controls. NGU may be caused by a number of different bacteria but more studies including a higher number of samples are needed for elucidation of the role of each species.


Assuntos
Adenoviridae , Bactérias Gram-Negativas , Herpesvirus Humano 1 , Herpesvirus Humano 2 , Uretrite , Urina , Adenoviridae/classificação , Adenoviridae/genética , Adulto , Bactérias Gram-Negativas/classificação , Bactérias Gram-Negativas/genética , Herpesvirus Humano 1/genética , Herpesvirus Humano 2/genética , Humanos , Masculino , Pessoa de Meia-Idade , Uretra/microbiologia , Uretra/virologia , Uretrite/microbiologia , Uretrite/urina , Uretrite/virologia , Urina/microbiologia , Urina/virologia
6.
Lab Chip ; 17(24): 4273-4282, 2017 12 05.
Artigo em Inglês | MEDLINE | ID: mdl-29116271

RESUMO

Three-dimensional (3D) in vitro models capturing both the structural and dynamic complexity of the in vivo situation are in great demand as an alternative to animal models. Despite tremendous progress in engineering complex tissue/organ models in the past decade, approaches that support the required freedom in design, detail and chemistry for fabricating truly 3D constructs have remained limited. Here, we report a stereolithographic high-resolution 3D printing technique utilizing poly(ethylene glycol) diacrylate (PEGDA, MW 700) to manufacture diffusion-open and mechanically stable hydrogel constructs as self-contained chips, where confined culture volumes are traversed and surrounded by perfusable vascular-like networks. An optimized resin formulation enables printing of hydrogel chips holding perfusable microchannels with a cross-section as small as 100 µm × 100 µm, and the printed microchannels can be steadily perfused for at least one week. In addition, the integration of multiple independently perfusable and structurally stable channel systems further allows for easy combination of different bulk material volumes at exact relative spatial positions. We demonstrate this structural and material flexibility by embedding a highly compliant cell-laden gelatin hydrogel within the confines of a 3D printed resilient PEGDA hydrogel chip of intermediate compliance. Overall, our proposed strategy represents an automated, cost-effective and high resolution technique to manufacture complex 3D constructs containing microfluidic perfusion networks for advanced in vitro models.


Assuntos
Técnicas de Cultura de Células/instrumentação , Hidrogéis/química , Técnicas Analíticas Microfluídicas/instrumentação , Impressão Tridimensional , Tecidos Suporte , Desenho de Equipamento , Células Endoteliais da Veia Umbilical Humana , Humanos , Modelos Biológicos , Perfusão
7.
Bioresour Technol ; 245(Pt A): 714-723, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28917107

RESUMO

The objective of this study was to delineate the effects of different coffee processing residues on the anaerobic microbes and corresponding digestion performance. The results elucidated that mucilage-rich feed enhanced the accumulation of methanogens, which consequently led to better digestion performance of biogas production. Fifty percent more methane and up to 3 times more net energy (heat and electricity) output were achieved by the digestion of the mucilage-rich feed (M3). The microbial community and statistical analyses further elucidated that different residues in the feed had significant impact on microbial distribution and correspondingly influenced the digestion performance.


Assuntos
Biocombustíveis , Café , Metano , Anaerobiose , Reatores Biológicos , Digestão , Euryarchaeota
8.
JAMA Dermatol ; 153(9): 897-905, 2017 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-28538949

RESUMO

Importance: Although the pathogenesis of hidradenitis suppurativa (HS) remains enigmatic, several factors point to potential involvement of the cutaneous microbiome. Insight into the cutaneous microbiome in HS using next-generation sequencing may provide novel data on the microbiological diversity of the skin. Objective: To investigate the follicular skin microbiome in patients with HS and in healthy controls. Design, Setting, and Participants: This case-control study obtained punch biopsy specimens from patients with HS (lesional and nonlesional) and healthy controls between October 1, 2014, and August 1, 2016. Data were analyzed from March to November 2016. Patients with HS were recruited from the Department of Dermatology, Zealand University Hospital, Roskilde, Denmark. Biopsy specimens were analyzed at the Department of Microbiology and Infection Control, Statens Serum Institut, Copenhagen, Denmark. None of the participants received any antibiotics (systemic or topical therapy) within 1 month before the study. In patients with HS, biopsy specimens were obtained from lesional skin (axilla or groin) and nonlesional skin. Only nodules containing at least 1 visible hair follicle were biopsied. Biopsy specimens from healthy controls were obtained from the axilla only. Main Outcomes and Measures: The different microbiomes were investigated using next-generation sequencing targeting 16S and 18S ribosomal RNA. Results: The skin microbiome was characterized in 30 patients with HS (mean [SD] age, 46.9 [14.0] years; 19 [63% female]) and 24 healthy controls (mean [SD] age, 32.2 [12.0] years; 13 [54% female]). The next-generation sequencing data provided a previously unreported (to our knowledge) characterization of the skin microbiome in HS. The study demonstrated that the microbiome in HS differs significantly from that in healthy controls in lesional and nonlesional skin. Overall, the following 5 microbiome types were identified: Corynebacterium species (type I), Acinetobacter and Moraxella species (type II), Staphylococcus epidermidis (type III), Porphyromonas and Peptoniphilus species (type IV), and Propionibacterium acnes (type V). In lesional skin, microbiome types consisted predominantly of type I or type IV. Microbiome type IV was not detected in healthy controls. Several taxa, including Propionibacterium, showed a significantly higher relative abundance in healthy controls vs HS skin, indicating that Propionibacterium may be part of the pathogenesis in HS. Conclusions and Relevance: The study findings suggest a link between a dysbiotic cutaneous microbiome and HS.


Assuntos
Hidradenite Supurativa/microbiologia , Microbiota , Pele/microbiologia , Adulto , Bactérias/genética , Estudos de Casos e Controles , DNA Bacteriano/análise , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Análise de Sequência de DNA
9.
Clin Rehabil ; 31(12): 1674-1683, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28523990

RESUMO

OBJECTIVE: The purpose of this study is to produce insight in the explanations for declining pulmonary rehabilitation given by patients with chronic obstructive pulmonary disease. SETTING: The participants were recruited from a hospital in Denmark, among patients hospitalized due to an exacerbation of chronic obstructive pulmonary disease and among stable patients attending an outpatient clinic. PARTICIPANTS: Patients who decline participation in offered pulmonary rehabilitation, who speak Danish, who are able to give informed consent and to participate in a 1-hour interview. METHOD: The research question was answered through interviews with 19 patients. DATA ANALYSIS: The interviews were recorded and transcribed verbatim. The transcripts were analyzed using inductive content analysis. The transcripts were condensed; categories were developed providing different types of explanations for declining pulmonary rehabilitation. Each category was named using a content characteristic word. RESULTS: This study shows that some patients do not remember or recall that they have been offered pulmonary rehabilitation during hospitalization. Especially the oldest patients perceive themselves to be too frail from chronic obstructive pulmonary disease, comorbidity or multimorbidity. The male patients tend to find pulmonary rehabilitation irrelevant because they do not consider themselves ill. Furthermore, the study shows that pulmonary rehabilitation is perceived to be time-consuming and conflicting with daily activities. CONCLUSIONS: Patients decline pulmonary rehabilitation because the intervention does not fit their perception of health and because they find that participation in the program may collide with priorities and daily activities.


Assuntos
Doença Pulmonar Obstrutiva Crônica/psicologia , Doença Pulmonar Obstrutiva Crônica/reabilitação , Recusa do Paciente ao Tratamento/psicologia , Idoso , Feminino , Humanos , Masculino , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Pesquisa Qualitativa , Fatores Socioeconômicos
10.
J Med Chem ; 60(1): 441-457, 2017 01 12.
Artigo em Inglês | MEDLINE | ID: mdl-28005385

RESUMO

Ionotropic glutamate receptor antagonists are valuable tool compounds for studies of neurological pathways in the central nervous system. On the basis of rational ligand design, a new class of selective antagonists, represented by (2S,4R)-4-(2-carboxyphenoxy)pyrrolidine-2-carboxylic acid (1b), for cloned homomeric kainic acid receptors subtype 1 (GluK1) was attained (Ki = 4 µM). In a functional assay, 1b displayed full antagonist activity with IC50 = 6 ± 2 µM. A crystal structure was obtained of 1b when bound in the ligand binding domain of GluK1. A domain opening of 13-14° was seen compared to the structure with glutamate, consistent with 1b being an antagonist. A structure-activity relationship study showed that the chemical nature of the tethering atom (C, O, or S) linking the pyrrolidine ring and the phenyl ring plays a key role in the receptor selectivity profile and that substituents on the phenyl ring are well accommodated by the GluK1 receptor.


Assuntos
Prolina/antagonistas & inibidores , Pirrolidinas/farmacologia , Receptores Ionotrópicos de Glutamato/antagonistas & inibidores , Receptores de Ácido Caínico/antagonistas & inibidores , Cristalografia por Raios X , Desenho de Fármacos , Concentração Inibidora 50 , Pirrolidinas/química , Relação Estrutura-Atividade
11.
PLoS One ; 11(8): e0160331, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27509169

RESUMO

Staphylococcus aureus is presently regarded as an emerging zoonotic agent due to the spread of specific methicillin-resistant S. aureus (MRSA) clones in pig farms. Studying the microbiota can be useful for the identification of bacteria that antagonize such opportunistic veterinary and zoonotic pathogen in animal carriers. The aim of this study was to determine whether the nasal microbiome of pig S. aureus carriers differs from that of non-carriers. The V3-V5 region of the 16S rRNA gene was sequenced from nasal swabs of 44 S. aureus carriers and 56 non-carriers using the 454 GS FLX titanium system. Carriers and non-carriers were selected on the basis of quantitative longitudinal data on S. aureus carriage in 600 pigs sampled at 20 Danish herds included in two previous studies in Denmark. Raw sequences were analysed with the BION meta package and the resulting abundance matrix was analysed using the DESeq2 package in R to identify operational taxonomic units (OTUs) with differential abundance between S. aureus carriers and non-carriers. Twenty OTUs were significantly associated to non-carriers, including species with known probiotic potential and antimicrobial effect such as lactic acid-producing isolates described among Leuconostoc spp. and some members of the Lachnospiraceae family, which is known for butyrate production. Further 5 OTUs were significantly associated to carriage, including known pathogenic bacteria such as Pasteurella multocida and Klebsiella spp. Our results show that the nasal microbiome of pigs that are not colonized with S. aureus harbours several species/taxa that are significantly less abundant in pig carriers, suggesting that the nasal microbiota may play a role in the individual predisposition to S. aureus nasal carriage in pigs. Further research is warranted to isolate these bacteria and assess their possible antagonistic effect on S. aureus for the pursuit of new strategies to control MRSA in pig farming.


Assuntos
Microbiota , Cavidade Nasal/microbiologia , Staphylococcus aureus/genética , Animais , Firmicutes/genética , Firmicutes/crescimento & desenvolvimento , Firmicutes/isolamento & purificação , Genótipo , Humanos , Klebsiella/genética , Klebsiella/crescimento & desenvolvimento , Klebsiella/isolamento & purificação , Leuconostoc/genética , Leuconostoc/crescimento & desenvolvimento , Leuconostoc/isolamento & purificação , Staphylococcus aureus Resistente à Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/crescimento & desenvolvimento , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Pasteurella multocida/genética , Pasteurella multocida/crescimento & desenvolvimento , Pasteurella multocida/isolamento & purificação , Fenótipo , RNA Ribossômico 16S/química , RNA Ribossômico 16S/genética , RNA Ribossômico 16S/metabolismo , Análise de Sequência de DNA , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus aureus/isolamento & purificação , Suínos
12.
J Org Chem ; 81(21): 10366-10375, 2016 11 04.
Artigo em Inglês | MEDLINE | ID: mdl-27441647

RESUMO

The scalable synthesis of cyclic enecarbamates and their use as convenient precursors of α,ß-unsaturated N-acyl iminium ions is reported. The newly developed route overcomes synthetic and reactivity difficulties in previously reported methods, is readily scaled up, and proceeds through stable intermediates suitable for long-term storage if required. Preliminary investigations probing the reactivity of cyclic α,ß-unsaturated N-acyl iminium ions as dienophiles in Diels-Alder reactions and electrophilic alkylating agents are described. In the presence of Lewis and Brønsted acids, iminium precursor 22a underwent efficient Diels-Alder cycloaddition with a range of simple and complex dienes, culminating in the synthesis of 6,6-spirocyclic ring systems possessing the same relative stereochemistry as the spirocyclic imine present in the marine natural product gymnodimine 1.

13.
Cytotherapy ; 18(9): 1187-96, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27424146

RESUMO

BACKGROUND AIMS: Maturation of dendritic cells (DCs) induces their homing from peripheral to lymphatic tissues guided by CCL21. However, in vitro matured human monocyte-derived DC cancer vaccines injected intradermally migrate poorly to lymph nodes (LNs). In vitro maturation protocols generate DCs with high (type 1 DCs) or low (prostaglandin E2 [PGE2]-DCs) autocrine CCL19 levels, which may potentially interfere with LN homing of DCs. METHODS: Employing a three-dimensional (3D) chemotaxis assay, chemokine competition/desensitization studies and short interfering RNA (siRNA) against CCL19, we analyzed the effect of autocrine CCL19 on in vitro migration of human DCs toward CCL21. RESULTS: Using human monocyte-derived DCs in a 3D chemotaxis assay, we are the first to demonstrate that CCL19 more potently induces directed migration of human DCs compared with CCL21. When comparing migration of type 1 DCs and PGE2-DCs, migration of type 1 DCs was strikingly impaired compared with PGE2-DCs, but only toward low concentrations of CCL21. When type 1 DCs were cultured overnight in fresh culture medium (reducing autocrine CCL19 levels), a rescuing effect was observed on migration toward low concentrations of CCL21 in a 3D chemotaxis assay. Finally pre-incubation with CCL19 negatively affected PGE2-DC migration, whereas silencing of CCL19 by siRNA improved type 1 DC migration. Importantly, in both cases, the effect was observed only at low concentrations of CCL21. CONCLUSIONS: Our results demonstrate that autocrine CCL19 negatively affects DC migratory potential toward CCL21, the potency difference between CCL19 and CCL21 being the underlying cause. CCL19 secretion level of in vitro matured DCs is an important indicator of DC vaccine homing potential.


Assuntos
Quimiocina CCL19/metabolismo , Células Dendríticas/citologia , Movimento Celular , Células Cultivadas , Quimiocina CCL19/genética , Quimiocina CCL19/farmacologia , Quimiocina CCL21/metabolismo , Quimiotaxia , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Dinoprostona/metabolismo , Humanos , Masculino , Monócitos/citologia
14.
PLoS One ; 11(6): e0157046, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27276347

RESUMO

PCR amplification and sequencing of phylogenetic markers, primarily Small Sub-Unit ribosomal RNA (SSU rRNA) genes, has been the paradigm for defining the taxonomic composition of microbiomes. However, 'universal' SSU rRNA gene PCR primer sets are likely to miss much of the diversity therein. We sequenced a library comprising purified and reverse-transcribed SSU rRNA (RT-SSU rRNA) molecules from the canine oral microbiome and compared it to a general bacterial 16S rRNA gene PCR amplicon library generated from the same biological sample. In addition, we have developed BIONmeta, a novel, open-source, computer package for the processing and taxonomic classification of the randomly fragmented RT-SSU rRNA reads produced. Direct RT-SSU rRNA sequencing revealed that 16S rRNA molecules belonging to the bacterial phyla Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria and Spirochaetes, were most abundant in the canine oral microbiome (92.5% of total bacterial SSU rRNA). The direct rRNA sequencing approach detected greater taxonomic diversity (1 additional phylum, 2 classes, 1 order, 10 families and 61 genera) when compared with general bacterial 16S rRNA amplicons from the same sample, simultaneously provided SSU rRNA gene inventories of Bacteria, Archaea and Eukarya, and detected significant numbers of sequences not recognised by 'universal' primer sets. Proteobacteria and Spirochaetes were found to be under-represented by PCR-based analysis of the microbiome, and this was due to primer mismatches and taxon-specific variations in amplification efficiency, validated by qPCR analysis of 16S rRNA amplicons from a mock community. This demonstrated the veracity of direct RT-SSU rRNA sequencing for molecular microbial ecology.


Assuntos
Bactérias , Sequenciamento de Nucleotídeos em Larga Escala , Microbiota/genética , Boca/microbiologia , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Animais , Bactérias/classificação , Bactérias/genética , Cães
15.
Methods Mol Biol ; 1428: 151-61, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27236798

RESUMO

Electroporation is well established for transient mRNA transfection of many mammalian cells, including immune cells such as dendritic cells used in cancer immunotherapy. Therapeutic application requires methods to efficiently electroporate and transfect millions of immune cells in a fast process with high cell survival. Continuous flow of suspended dendritic cells through a channel incorporating spatially separated microporous meshes with a synchronized electrical pulsing sequence can yield dendritic cell transfection rates of >75 % with survival rates of >90 %. This chapter describes the instrumentation and methods needed for the efficient transfection by electroporation of millions of dendritic cells in one continuous flow process.


Assuntos
Células Dendríticas/citologia , Eletroporação/instrumentação , Eletroporação/métodos , RNA Mensageiro/metabolismo , Proliferação de Células , Sobrevivência Celular , Células Dendríticas/metabolismo , Técnicas de Transferência de Genes , Proteínas de Fluorescência Verde/genética , Humanos , Transfecção
16.
Adv Healthc Mater ; 5(2): 244-54, 2016 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-26619161

RESUMO

Stable and low-cost multiplexed drug sensitivity assays using small volumes of cells or tissue are in demand for personalized medicine, including patient-specific combination chemotherapy. Spatially defined projected light photopolymerization of hydrogels with embedded active compounds is introduced as a flexible and cost-efficient method for producing multiplexed dosing assays. The high spatial resolution of light projector technology defines multiple compound doses by the volume of individual compound-embedded hydrogel segments. Quantitative dosing of multiple proteins with a dynamic range of 1-2 orders of magnitude is demonstrated using fluorescently labeled albumins. The hydrogel matrix results from photopolymerization of low-cost poly(ethylene glycol) diacrylates (PEGDA), and tuning of the PEGDA composition enables fast complete dosing of all tested species. Dosing of hydrophilic and hydrophobic compounds is demonstrated using two first-line chemotherapy regimens combining oxaliplatin, SN-38, 5-fluorouracil, and folinic acid, with each compound being dosed from a separate light-defined hydrogel segment. Cytotoxicity studies using a colorectal cancer cell line show equivalent effects of dissolved and released compounds. Further control of the dosing process is demonstrated by liposomal encapsulation of oxaliplatin, stable embedding of the liposomes in hydrogels for more than 3 months, and heat-triggered complete release of the loaded oxaliplatin.


Assuntos
Bioensaio/métodos , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Antineoplásicos/farmacologia , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Liberação Controlada de Fármacos , Estabilidade de Medicamentos , Fluoruracila/farmacologia , Humanos , Leucovorina/farmacologia , Lipossomos/química , Compostos Organoplatínicos/farmacologia , Oxaliplatina , Polietilenoglicóis/química , Temperatura , Fatores de Tempo
17.
Scand J Caring Sci ; 30(3): 432-41, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26426088

RESUMO

AIM: This paper aimed to suggest possible answers to the question: Why do patients with COPD decline pulmonary rehabilitation (PR)? METHOD: The study is a metasynthesis inspired by Noblit of the existing qualitative research on the area. The data were collected during 2014. Six studies were found through a systematic literature search in relevant databases. In these six studies, 65 persons were identified as decliners of PR. Four themes were identified from these studies. RESULTS: The themes identified were as follows: the referral process, transport problems, perception of health and other obligations or priorities. The problems with the referral of patients relate to different areas: the referring health professional's conviction and commitment, and the patients' understanding of the referral. It seems that various transport problems cause decline, for example long distance to the PR centre or the expenses of getting back and forth. Perceptions of health cause decline. Decliners feel too sick to join PR or do not identify themselves as a sick person, and do not want undertake the 'patient role'. Other obligations or priorities such as work, family obligations and vacations are prioritised on behalf of PR causing decline. CONCLUSION: The studies included show patients' rational accounts and reflections on declining PR. The included studies tend to describe accounts for deselection of PR in relation to the preferences and beliefs of the patients rather than including the social and economic variables framing the behaviour and choices of the patients.


Assuntos
Aceitação pelo Paciente de Cuidados de Saúde , Doença Pulmonar Obstrutiva Crônica/reabilitação , Humanos
18.
Nucleic Acids Res ; 43(21): e139, 2015 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-26163061

RESUMO

Nucleic acid aptamer selection by systematic evolution of ligands by exponential enrichment (SELEX) has shown great promise for use in the development of research tools, therapeutics and diagnostics. Typically, aptamers are identified from libraries containing up to 10(16) different RNA or DNA sequences by 5-10 rounds of affinity selection towards a target of interest. Such library screenings can result in complex pools of many target-binding aptamers. New high-throughput sequencing techniques may potentially revolutionise aptamer selection by allowing quantitative assessment of the dynamic changes in the pool composition during the SELEX process and by facilitating large-scale post-SELEX characterisation. In the present study, we demonstrate how high-throughput sequencing of SELEX pools, before and after a single round of branched selection for binding to different target variants, can provide detailed information about aptamer binding sites, preferences for specific target conformations, and functional effects of the aptamers. The procedure was applied on a diverse pool of 2'-fluoropyrimidine-modified RNA enriched for aptamers specific for the serpin plasminogen activator inhibitor-1 (PAI-1) through five rounds of standard selection. The results demonstrate that it is possible to perform large-scale detailed characterisation of aptamer sequences directly in the complex pools obtained from library selection methods, thus without the need to produce individual aptamers.


Assuntos
Aptâmeros de Nucleotídeos/química , Sequenciamento de Nucleotídeos em Larga Escala , Técnica de Seleção de Aptâmeros , Sítios de Ligação , Ligantes , Mutação , Conformação de Ácido Nucleico , Inibidor 1 de Ativador de Plasminogênio/genética , Inibidor 1 de Ativador de Plasminogênio/metabolismo , RNA/química , Alinhamento de Sequência , Análise de Sequência de RNA
19.
J Med Chem ; 58(15): 6131-50, 2015 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-26200741

RESUMO

Herein we describe the first structure-activity relationship study of the broad-range iGluR antagonist (2S,3R)-3-(3-carboxyphenyl)pyrrolidine-2-carboxylic acid (1) by exploring the pharmacological effect of substituents in the 4, 4', or 5' positions and the bioisosteric substitution of the distal carboxylic acid for a phosphonic acid moiety. Of particular interest is a hydroxyl group in the 4' position 2a which induced a preference in binding affinity for homomeric GluK3 over GluK1 (Ki = 0.87 and 4.8 µM, respectively). Two X-ray structures of ligand binding domains were obtained: 2e in GluA2-LBD and 2f in GluK1-LBD, both at 1.9 Å resolution. Compound 2e induces a D1-D2 domain opening in GluA2-LBD of 17.3-18.8° and 2f a domain opening in GluK1-LBD of 17.0-17.5° relative to the structures with glutamate. The pyrrolidine-2-carboxylate moiety of 2e and 2f shows a similar binding mode as kainate. The 3-carboxyphenyl ring of 2e and 2f forms contacts comparable to those of the distal carboxylate in kainate.


Assuntos
Antagonistas de Aminoácidos Excitatórios/química , Antagonistas de Aminoácidos Excitatórios/farmacologia , Pirrolidinas/farmacologia , Receptores Ionotrópicos de Glutamato/antagonistas & inibidores , Cristalografia por Raios X , Modelos Moleculares , Relação Estrutura-Atividade
20.
BMC Vet Res ; 11: 139, 2015 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-26099928

RESUMO

BACKGROUND: In recent years, new neonatal porcine diarrhoea (NNPD) of unknown aetiology has emerged in Denmark. NNPD affects piglets during the first week of life and results in impaired welfare, decreased weight gain, and in the worst-case scenario death. Commonly used preventative interventions such as vaccination or treatment with antibiotics, have a limited effect on NNPD. Previous studies have investigated the clinical manifestations, histopathology, and to some extent, microbiological findings; however, these studies were either inconclusive or suggested that Enterococci, possibly in interaction with Escherichia coli, contribute to the aetiology of NNPD. This study examined ileal and colonic luminal contents of 50 control piglets and 52 NNPD piglets by means of the qPCR-based Gut Microbiotassay and 16 samples by 454 sequencing to study the composition of the bacterial gut microbiota in relation to NNPD. RESULTS: NNPD was associated with a diminished quantity of bacteria from the phyla Actinobacteria and Firmicutes while genus Enterococcus was more than 24 times more abundant in diarrhoeic piglets. The number of bacteria from the phylum Fusobacteria was also doubled in piglets suffering from diarrhoea. With increasing age, the gut microbiota of NNPD affected piglet and control piglets became more diverse. Independent of diarrhoeic status, piglets from first parity sows (gilts) possessed significantly more bacteria from family Enterobacteriaceae and species E. coli, and fewer bacteria from phylum Firmicutes. Piglets born to gilts had 25 times higher odds of having NNPD compared with piglets born to multiparous sows. Finally, the co-occurrence of genus Enterococcus and species E. coli contributed to the risk of having NNPD. CONCLUSION: The results of this study support previous findings that points towards genus Enterococcus and species E. coli to be involved in the pathogenesis of NNPD. Moreover, the results indicate that NNPD is associated with a disturbed bacterial composition and larger variation between the diarrhoeic piglets.


Assuntos
Animais Recém-Nascidos , Bactérias/isolamento & purificação , Diarreia/veterinária , Trato Gastrointestinal/microbiologia , Doenças dos Suínos/etiologia , Animais , Bactérias/classificação , Biologia Computacional , Diarreia/etiologia , Análise de Componente Principal , Suínos , Doenças dos Suínos/microbiologia
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