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1.
Chem Commun (Camb) ; 55(77): 11615-11618, 2019 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-31501837

RESUMO

To date, implementation of renewable DNA circuits remains challenging due to issues including reactant depletion and waste accumulation. Herein we simultaneously addressed both issues through nicking enzyme-assisted waste-to-reactant transformation. As a proof-of-concept, a renewable entropy-driven catalytic DNA circuit was implemented, exhibiting a good renewability when replenishing fuel.


Assuntos
DNA Catalítico/química , Endonucleases/química , Redes Reguladoras de Genes , Pareamento Incorreto de Bases , Catálise , Entropia , Corantes Fluorescentes/química , Cinética , Conformação de Ácido Nucleico , Estudo de Prova de Conceito , Espectrometria de Fluorescência
2.
Chem Commun (Camb) ; 55(80): 12036-12039, 2019 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-31531454

RESUMO

While commercially available suncare products are effective at absorbing ultraviolet (UV)-light, recent studies indicate systemic toxicities associated with many traditional chemical and physical UV-filters. We demonstrate the application of xanthommatin, a biochrome present in arthropods and cephalopods, as an alternative chemical UV-filter that is cytocompatible while maintaining its photostability and photoprotective properties.


Assuntos
Antioxidantes/farmacologia , Oxazinas/farmacologia , Pele/efeitos da radiação , Protetores Solares/farmacologia , Xantenos/farmacologia , Animais , Antioxidantes/química , Sobrevivência Celular/efeitos dos fármacos , Dano ao DNA , Dimetilpolisiloxanos/química , Humanos , Camundongos , Células NIH 3T3 , Oxazinas/química , Estudo de Prova de Conceito , Pele/citologia , Protetores Solares/química , Raios Ultravioleta , Xantenos/química
3.
Chem Commun (Camb) ; 55(67): 9971-9974, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31367709

RESUMO

Photodynamic therapy (PDT) is a clinically approved cancer treatment that uses light, oxygen and a photosensitizer to produce localized reactive oxygen species (ROS). Due to the short lifetime of ROS, the location of the photosensitizer in the cell is believed to be the key determinant governing the outcome of PDT. To explore the effect of direct association between a photosensitizer and DNA a well know DNA-binding dye, DAPI, was converted into a photosensitizer. Br-DAPI - unlike native DAPI - upon irradiation produces ROS. We demonstrate that the ROS are only effective in inducing dsDNA breaks when Br-DAPI is bound to DNA. In cancer cells (A549) Br-DAPI causes rapid light dependent cell death. This work supports the design of photosensitizers which bind with high affinity to the DNA of target cells for potentially more effective PDT.


Assuntos
Bromo/química , DNA/química , Indóis/química , Fármacos Fotossensibilizantes/química , Células A549 , Apoptose/efeitos dos fármacos , Apoptose/efeitos da radiação , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Dano ao DNA , Corantes Fluorescentes/química , Humanos , Luz , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacologia , Estudo de Prova de Conceito , Espécies Reativas de Oxigênio/química , Espécies Reativas de Oxigênio/metabolismo
4.
Chem Commun (Camb) ; 55(70): 10380-10383, 2019 Aug 27.
Artigo em Inglês | MEDLINE | ID: mdl-31397448

RESUMO

A strategy for the photoelectrochemical detection of miRNA with ultra-low background noise was developed using tungsten diselenide-cysteine-dopamine (WSe2/Cys/DA) as a nanoprobe coupled with mismatched catalytic hairpin assembly target recycling. A superior detection limit of 3.3 aM toward miRNA-221 was achieved.


Assuntos
Cisteína/química , Dopamina/química , Técnicas Eletroquímicas/métodos , MicroRNAs/análise , Sondas Moleculares/química , Nanoestruturas , Processos Fotoquímicos , Selênio/química , Tungstênio/química , Técnicas Biossensoriais , Catálise , Humanos , Limite de Detecção , MicroRNAs/sangue , Estudo de Prova de Conceito
6.
Chem Commun (Camb) ; 55(73): 10952-10955, 2019 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-31441915

RESUMO

Triggering antibody-mediated innate immune mechanisms to kill cancer cells is an attractive therapeutic avenue. In this context, recruitment of endogenous antibodies to the cancer cell surface could be a viable alternative to the use of monoclonal antibodies. We report on antibody-recruiting polymers containing multiple antibody-binding hapten motifs and cyclooctynes that can covalently conjugate to azides introduced onto the glycocalyx of cancer cells by metabolic labeling with azido sugars.


Assuntos
Resinas Acrílicas/química , Anticorpos/imunologia , Azidas/metabolismo , Dinitrobenzenos/imunologia , Hexosaminas/metabolismo , Resinas Acrílicas/síntese química , Animais , Azidas/química , Linhagem Celular Tumoral , Química Click , Reação de Cicloadição , Ciclo-Octanos/síntese química , Ciclo-Octanos/química , Dinitrobenzenos/síntese química , Dinitrobenzenos/química , Fluorescência , Corantes Fluorescentes/química , Glicocálix/metabolismo , Hexosaminas/química , Humanos , Camundongos , Microscopia Confocal/métodos , Microscopia de Fluorescência/métodos , Estudo de Prova de Conceito , Esferoides Celulares/metabolismo
7.
Zhongguo Zhong Yao Za Zhi ; 44(13): 2657-2661, 2019 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-31359673

RESUMO

Target identification is an important prerequisite for the study of medicine action mechanism. Currently,drug target identification is mostly based on various cell models in vitro. However,the growth microenvironment,nutrition metabolism,biological properties as well as functions are quite different between in vitro cell culture and physiological environment in vivo; wherefore,it is a challenging scientific issue to establish an effective method for identifying drug targets in vivo condition. In this study,we successfully prepared a kind of magnetic nanoparticles( MNPs) which can be chemically modified by the hydroxyl structure of natural bioactive compound echinacoside( ECH) via the epoxy group label on the surface of MNPs. Therefore,organ-selective and recoverable nanoscale target-recognizing particles were prepared. We then intravenously injected the ECH-binding MNPs into rats and distributed them to specific organs in vivo. After cell endocytosis,ECH-binding MNPs captured target proteins in situ for further analysis. Based on this method,we discovered several potential target proteins in the spleen lysates for ECH,and preliminarily clarified the immuno-regulation mechanism of ECH. Collectively,our strategy developed a proof-of-concept technology using nanoparticles for in vivo target identification,and also provided a feasible approach for drug target prediction and pharmacological mechanism exploration.


Assuntos
Sistemas de Liberação de Medicamentos , Nanopartículas de Magnetita , Medicina Tradicional Chinesa , Animais , Endocitose , Glicosídeos/análise , Magnetismo , Estudo de Prova de Conceito , Ratos
8.
Medicine (Baltimore) ; 98(28): e16347, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31305424

RESUMO

Blood pressure variability (BPV) is an independent cardiovascular risk factor in hypertensive patients. The best method for quantifying BPV is still an object of debate. The existence of different BPV patterns, particularly age and arterial stiffness related, is postulated. Our aims were:Cross-sectional study in 108 elderly hypertensive hospitalized patients. Each patient underwent blood pressure measurements with 5 different modalities: 24 hour BP and pulse wave velocity (PWV) monitoring (24hBPM), measurement by nurses or physicians, self-measurement and beat-to-beat monitoring. Differences between maximum and minimum values (ΔBP), averages of the absolute differences between consecutive values (ARV) and coefficients of variation (CV) were calculated.ΔBP showed the wider values' dispersion (Δ systolic blood pressure (SBP): 66.4 ±â€Š22.9 and Δ diastolic blood pressure [DBP]: 45.0 ±â€Š13.5 mmHg). ARV and CV were highest with nurses' measurements (SBP-ARV 9.2 ±â€Š6.2; DBP-ARV 6.9 ±â€Š5.2; SBP-CV 7.6 ±â€Š5.3; DBP-CV 9.6 ±â€Š5.5). The strongest correlation was found comparing physicians' SBP measurements and 24hBPM ARVs (R2 0.23, P <.05). 24hBPM ΔSBP in a multivariate analysis was significantly associated with age (ß -3.85, SE 0.83; P <.001) and PWV (ß 20.29, SE 3.70; P <.001). Calcium antagonists were associated with a lower ΔSBP (ß -14.6, SE 6.1, P <.05) while diuretics and alpha-blockers with a significant increase (ß 14.4 SE 5.4, P <.01; ß 26.9 SE 11.7, P <.05).Age, PWV, diuretics, alpha-blockers, but also measurements obtained by nurses, increase BP variability while calcium antagonists reduce it. BP profiles in elderly in-hospital patients potentially provide important information; they should, however, be interpreted cautiously.


Assuntos
Determinação da Pressão Arterial/métodos , Pressão Sanguínea , Hipertensão/diagnóstico , Idoso de 80 Anos ou mais , Pressão Sanguínea/fisiologia , Estudos Transversais , Feminino , Hospitalização , Humanos , Hipertensão/fisiopatologia , Hipertensão/terapia , Masculino , Estudo de Prova de Conceito
9.
Medicine (Baltimore) ; 98(26): e15917, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31261498

RESUMO

Acute onset neurological symptoms evoked by traumatic, surgical, or emotional events in Wilson disease (WD) have never been reported and its clinical characteristics are unclear.We aimed to summarize the clinical characteristics of a special WD whose neurological symptoms acutely developed after traumatic, surgical, or emotional events.Retrospective pilot study.Thirty-one patients who had acute onset neurological symptom as an initial presentation of WD or a new presentation of hepatic WD after mild trauma, surgery, or emotional events were retrospectively studied. All patients were followed for half to 1 year after regular anti-copper treatment.The averaged latency for neurological symptom presentation was 2.79 ±â€Š1.21 hours. The most frequent neurological symptoms were tremor (74%) and basal ganglia (BG) lesions were detected on magnetic resonance imaging in all patients. Lesions in other regions were much less frequently detected. Neurological symptom score and its recovery after treatment were correlated with lesion location: BG area and BG plus other brain areas. Neurological symptoms improved in 21 patients who received timely anti-copper treatment but continued to deteriorate in 6 patients who did not accept regular anti-copper treatment for delayed diagnosis.A diagnosis of WD should be considered when adolescents or adults experience acute presentation of extrapyramidal systems after traumatic, surgical, or emotional stimulation. Timely anti-copper therapy usually gives rise to an excellent prognosis.


Assuntos
Emoções , Degeneração Hepatolenticular/epidemiologia , Degeneração Hepatolenticular/fisiopatologia , Complicações Pós-Operatórias/epidemiologia , Estresse Psicológico/complicações , Estresse Psicológico/epidemiologia , Adolescente , Adulto , Encéfalo/diagnóstico por imagem , Criança , Estudos Transversais , Feminino , Seguimentos , Degeneração Hepatolenticular/tratamento farmacológico , Humanos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Estudo de Prova de Conceito , Estudos Retrospectivos , Fatores de Tempo , Adulto Jovem
10.
Chem Commun (Camb) ; 55(58): 8402-8405, 2019 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-31257385

RESUMO

A proof-of-principle for the application of hemi-indigo derivatives as RNA binders with photocontrollable fluorescence is presented. The photoswitch binds to the human immunodeficiency virus type 1 (HIV-1) RNA with a significant light-up effect. The fluorescence of the RNA-bound ligand can be reversibly switched ON and OFF by light without destroying the ligand-RNA associates.


Assuntos
Corantes Fluorescentes/metabolismo , HIV-1/genética , Indóis/metabolismo , RNA/metabolismo , Fluorescência , Corantes Fluorescentes/química , Corantes Fluorescentes/efeitos da radiação , Indóis/química , Indóis/efeitos da radiação , Ligantes , Luz , Estudo de Prova de Conceito , RNA/genética , Elementos de Resposta , Estereoisomerismo
11.
Oncology ; 97(2): 59-74, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31261152

RESUMO

Chimeric antigen receptor (CAR) modified T-cell therapy, a unique platform technology highlighting precision medicine through utilization of molecular biology and cell-based therapeutics has shown unprecedented rates in patients with hematological malignancies such as acute lymphocyte leukemia, non-Hodgkin's lymphoma and multiple myeloma (MM). With the approval of CD19-targeted CAR T-cells by the Food and Drug Administration in acute lymphoblastic leukemia (ALL) and NHL, this technology is positioned for aggressive expansion to combination therapeutic opportunities and proof of principle towards utility in other malignant disorders. However, despite the impressive results seen with hematological malignancies, CAR T-cells have shown limited efficacy in solid tumors with several unsuccessful preclinical studies. Regardless, these attempts have provided us with a better understanding of the imminent challenges specific to solid tumors even if they have not so far led to expanded clinical treatment opportunities outside ALL/NHL/MM. This review summarizes our current understanding of CAR T-cell mechanism of action, while presenting the major limitations of CAR T-cell derived treatments in solid tumors. We further discuss recent findings and present new potential strategies to overcome the challenges facing solid tumor targeting by CAR T-cell platforms.


Assuntos
Imunoterapia Adotiva , Linfoma não Hodgkin/terapia , Mieloma Múltiplo/terapia , Leucemia-Linfoma Linfoblástico de Células Precursoras/terapia , Receptores de Antígenos Quiméricos/imunologia , Antígenos CD19/imunologia , Humanos , Estudo de Prova de Conceito
12.
Anal Chim Acta ; 1078: 112-118, 2019 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-31358208

RESUMO

Herein, an array-based in situ fluorescence assay is proposed for high-throughput analysis and localization of multiplex matrix metalloproteinases (MMPs) activities in cell monolayers and tissue sections. Five specific MMPs (MMP-2, -3, -7, -9, and -14) peptide substrates containing FAM/Dabcyl fluorescent resonance energy transfer (FRET) pair are directly spotted on the surface of cell monolayers or tissue sections, and hydrolyzed by localized MMPs, resulting in fluorescence recovery of FAM. MMPs activities are determined by the fluorescence intensity of stained cells/tissues due to the cellular internalization of peptide fragments with FAM moiety. We demonstrate that the array-based in situ fluorescence assay is suitable for identifying the MMPs expression patterns of cells, as well as determining the secreted MMPs activities in cell monolayer with high sensitivity (as low as hundreds of cells per square centimeter). The feasibility of the assay is further confirmed by evaluating inhibition potencies of six compounds toward five MMPs. Profiling of five MMPs activities in the localized parts of 32 thyroid tissues is performed without separation or extraction procedures, demonstrating the good practicality of the method.


Assuntos
Ensaios Enzimáticos/métodos , Metaloproteinases da Matriz/análise , Microscopia de Fluorescência/métodos , Peptídeos/metabolismo , Linhagem Celular Tumoral , Fluoresceínas/química , Fluorescência , Corantes Fluorescentes/química , Humanos , Hidrólise , Inibidores de Metaloproteinases de Matriz/farmacologia , Metaloproteinases da Matriz/metabolismo , Peptídeos/química , Estudo de Prova de Conceito , Glândula Tireoide/metabolismo
13.
Anal Chim Acta ; 1078: 151-160, 2019 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-31358213

RESUMO

Herein, we report a new signal amplification scheme for quantitative biochemical analysis based on gold nanoparticle (GNPs) catalyzed polymerization of transparent silane solution to milky white and turbid siloxane. Using immunoassay as a proof of concept, GNP labeled immunoprobe was used to bind captured antigen and catalyse the polymerization reaction allowing sensitive biochemical investigation. The polymerization reaction was optimized for standard 96 well polystyrene microtiter plates and we discovered that sodium lactate acts as an enhancer in the polymerization reaction as it reduces detection time to merely 30 min. The sensing strategy was applied to detection and quantification of Salmonella Typhimurium in water and egg samples and the platform showed excellent visibly quantifiable analytical response up to 100 cells mL-1. Furthermore, clinical utility and potential of the method was validated by detecting Vi capsular polysaccharide (Vi antigen) responsible for typhoidal Salmonellosis in human serum in sandwich format with a detection limit of 1 ng mL-1. The method serves as the first report towards nanoparticle triggered polymerization for development of rapid and low cost quantitative biochemical assay.


Assuntos
Ouro/química , Imunoensaio/métodos , Nanopartículas Metálicas/química , Polissacarídeos Bacterianos/sangue , Salmonella typhimurium/isolamento & purificação , Siloxanas/síntese química , Animais , Anticorpos/imunologia , Galinhas , Água Potável/microbiologia , Ovos/microbiologia , Contaminação de Alimentos/análise , Humanos , Limite de Detecção , Tamanho da Partícula , Polimerização , Polissacarídeos Bacterianos/imunologia , Estudo de Prova de Conceito , Salmonella typhimurium/imunologia , Silanos/química , Temperatura Ambiente
14.
Anal Chim Acta ; 1078: 182-188, 2019 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-31358218

RESUMO

Early detection of toxic proteases in food matrices plays a major role in preventing the occurrence of diseases as well as outbreaks. However, on-site detection of proteases, for instance, botulinum, anthrax and cholera in food matrices remains challenging due to their extremely low lethal dose levels. Here, we report a lateral flow assay (LFA) in a dipstick format for on-site visual detection of proteases in food matrices. The light chain of BoNT serotype A (BoNT/A LC) is used as a model system for validation of the proposed assay using magnetic beads conjugated to a synthetic peptide that provide a specific cleavage site for BoNT/A LC. Magnetic beads serve as both reporters for visual detection and as facilitators for sample clean-up, owing to the efficient magnetic separation protocol adopted. Digestion of the peptide substrate by BoNT/A LC for 5 h followed by the dipstick assay yields a reduction in color intensity of the test line on the dipstick compared to the control line obtained using an un-cleaved peptide substrate. Concentration dependent responses for the assay in carrot juice were obtained with a limit of detection (LOD) of 1 nM/2.5 nM (with/without amplification), also supported by RGB (ΔE) analysis, indicating the potential of the proposed methodology for on-site assaying of proteases in food matrices. Unlike typical affinity-based assays that yield a collective response for the active and inactive forms of the proteases, the proposed functional LFA targets only the active form, thereby enabling a more precise analysis for preventing potential false-positives. The proposed approach could be extended for detection of BoNT serotypes and other proteases in food matrices, upon utilizing appropriate substrates with specific cleavage sites.


Assuntos
Bioensaio/métodos , Toxinas Botulínicas Tipo A/análise , Sucos de Frutas e Vegetais/análise , Sequência de Aminoácidos , Anticorpos/imunologia , Toxinas Botulínicas Tipo A/química , Daucus carota/química , Limite de Detecção , Fenômenos Magnéticos , Peptídeos/química , Polietilenoglicóis/química , Estudo de Prova de Conceito , Estreptavidina/química
15.
Analyst ; 144(13): 3999-4005, 2019 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-31172144

RESUMO

Micrococcal nuclease (MNase) is an extracellular endonuclease of Staphylococcus aureus (S. aureus). It digests single stranded nucleic acid. The presence of MNase is the gold standard to identify S. aureus and its content. The present study reports the ultrahigh sensitive and selective fluorescence platform for MNase detection, designed and developed based on the surface energy transfer mechanism. A "proof of concept" is being developed based on monoclonal antibody-conjugated quantum dots (mAb-QDs), wherein mAb-QDs act as donors and graphene oxide (GO) acts as an acceptor. mAb-QDs in close proximity to GO undergo adsorption due to weak affinity between them and this results in fluorescence quenching by the transfer of surface energy from mAb-QDs to GO. During sensing, a much stronger affinity of mAb-QDs towards MNase inhibits the energy transfer to GO and this allows the regaining of fluorescence. Immobilized mAb-QDs on nitrocellulose membrane strips were fabricated and tested for "ON-OFF-ON" sensing of MNase. The limit of detection for fluorescence based assay and strips is found to be 0.3 ng mL-1 and 0.5 ng mL-1, respectively. The developed strips were applied on real samples for the detection of S. aureus.


Assuntos
Corantes Fluorescentes/química , Grafite/química , Nuclease do Micrococo/análise , Pontos Quânticos/química , Staphylococcus aureus/química , Anticorpos Monoclonais/química , Técnicas Biossensoriais/métodos , Colódio/química , Transferência Ressonante de Energia de Fluorescência/métodos , Limite de Detecção , Membranas Artificiais , Nanocompostos , Estudo de Prova de Conceito , Sensibilidade e Especificidade , Staphylococcus aureus/enzimologia
16.
BMC Neurol ; 19(1): 118, 2019 Jun 08.
Artigo em Inglês | MEDLINE | ID: mdl-31176377

RESUMO

BACKGROUND: In patients suffering from Chronic Inflammatory Demyelinating Polyneuropathy (CIDP) disease severity is assessed by Medical Research Counsil (MRC) Scale or Inflammatory Neuropathy Cause and Treatment (INCAT) disability score. However, none of these methods is appropriate to objectively assess muscle weakness or to detect very small subclinical changes. More objective and quantitative measures are needed in order to evaluate treatment efficiency or to detect subclinical affection of upper limps for early diagnosis. The goal of our study was to objectively quantify muscular weakness in CIDP patients with the non-invasive Quantitative Motor (Q-Motor) test of Grip Force Assessment (QGFA) as well as the Involuntary Movement Assessment (QIMA) and to search for differences between typical and atypical CIDP variants. In addition, we hypothesized that Q-Motor findings correlate with disease severity scales such as MRC or INCAT score. METHODS: In this cross-sectional exploratory proof-of-concept study subjects with confirmed diagnosis of typical or atypical CIDP were examined and compared to healthy controls (HC). For Q-Motor tests all subjects had to lift a device (250 g and 500 g) equipped with an electromagnetic sensor that measured grip force (GF) and three-dimensional changes in position and orientation. The measures "grip force variability" (GFV), "position index" (PI) and "orientation index" (OI) were provided to assess involuntary movements due to muscular weakness. RESULTS: 33 patients with CIDP and 28 HC were included. All measures were significantly elevated in CIDP patients for both devices in the right and left hand compared to healthy controls. Subgroup analysis revealed no differences between typical and atypical CIDP variants. INCAT score only weakly correlated with OI and PI. However, there was a stronger correlation between MRC and QIMA parameters in both hands. CONCLUSION: Q-Motor assessments were capable to objectively assess muscular weakness in CIDP. In particular, QIMA measures detected subclinical generalized muscle weakness even in patients with milder disability. Sensitivity and rater-independence of Q-Motor assessments support a further exploration of QIMA measures as potential endpoints for future clinical trials in CIDP.


Assuntos
Força da Mão/fisiologia , Debilidade Muscular , Polirradiculoneuropatia Desmielinizante Inflamatória Crônica , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Debilidade Muscular/diagnóstico , Debilidade Muscular/etiologia , Debilidade Muscular/fisiopatologia , Polirradiculoneuropatia Desmielinizante Inflamatória Crônica/complicações , Polirradiculoneuropatia Desmielinizante Inflamatória Crônica/fisiopatologia , Estudo de Prova de Conceito
17.
J Mass Spectrom ; 54(8): 676-683, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31240800

RESUMO

Detection of nucleic acids and single nucleotide polymorphisms (SNPs) is of pivotal importance in biology and medicine. Given that the biological effect of SNPs often is enhanced in combination with other SNPs, multiplexed SNP detection is desirable. We show proof of concept of the multiplexed detection of SNPs based on the template-directed native chemical ligation (NCL) of PNA-probes carrying a metal tag allowing detection using ICP-MS. For the detection of ssDNA oligonucleotides (30 bases), two probes, one carrying the metal tag and a second one carrying biotin for purification, are covalently ligated. The methodological limit of detection is of 29 pM with RSD of 6.7% at 50 pM (n = 5). Detection of SNPs is performed with the combination of two sets of reporter probes. The first probe set targets the SNP, and its yield is compared with a second set of probes targeting a neighboring sequence. The assay was used to simultaneously differentiate between alleles of three SNPs at 5-nM concentration.


Assuntos
Espectrometria de Massas/métodos , Ácidos Nucleicos/análise , Polimorfismo de Nucleotídeo Único , Sequência de Bases , Biotina/química , Cromatografia Líquida de Alta Pressão , Ligantes , Limite de Detecção , Metais/química , Oligonucleotídeos/análise , Estudo de Prova de Conceito , Estreptavidina/química
18.
Z Orthop Unfall ; 157(5): 501-509, 2019 Oct.
Artigo em Inglês, Alemão | MEDLINE | ID: mdl-31163514

RESUMO

BACKGROUND: The aim of the study is to identify those patients without improved pain sensation and functionality of the joint, thus with a potential necessity of re-intervention by using the evaluation of the PROMs (Oxford Hip Score, European Quality of Life Scale, EndoCert-Risk-Score). The task was to determine the value of the scores which represents the limit above/below, which a re-examination of the patient is necessary. Additionally, further secondary objectives such as influence of complications on scores, the percentage of questionnaire return and the quality of the provided data in the questionnaires were recorded to define minimal requirements on patient's response. MATERIAL AND METHODS: Members of the EndoCert certification committee developed a questionnaire for pre- (decision on treatment) and post-operative (discharge and 3-months follow-up) use. A total of 112 primary hip replacements and 25 hip revision arthroplasties, which were recruited from a centre of excellence for joint replacement (EndoProthetikZentrum der Maximalversorgung), were included in the study. RESULTS: The return of questionnaires amounted to 80%. Quality of data in the questionnaires declined with BMI (≥ 25), ASA classification (≥ 3), age (≥ 65) and depended on the time point questioning. Based on the EndoCert risk score there was a significant improvement in the experience of pain postoperatively compared to pre-operation (p < 0.05). However, the functionality of the joint, as measured as the use of walking frames or crutches as well as the need for help from others, did not improve. The results revealed that there was no correlation between the incidence of complications and a detrimental score. It was furthermore shown that patient specific attributes, e.g. age or BMI, or perioperative performance (duration of operation) did not influence the likelihood of a complication. CONCLUSION: It was shown that Oxford Hip Score and EQ-5D were not negatively influenced by complication during hospital stay. The evaluation of the EndoCert risk score showed not improvement between pre- and post-operative values regarding function but revealed a significant pain relief. A further study is intended to evaluate the intensity of pain after three months as part of an extended systemic follow-up. This could provide further insights whether the EndoCert risk score is a suitable tool to identify unsuccessfully treated patients.


Assuntos
Artroplastia de Quadril , Medidas de Resultados Relatados pelo Paciente , Artroplastia de Quadril/normas , Pesquisas sobre Serviços de Saúde , Instalações de Saúde/normas , Humanos , Estudo de Prova de Conceito , Qualidade de Vida , Resultado do Tratamento
19.
Protein Pept Lett ; 26(9): 691-701, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31215364

RESUMO

BACKGROUND: Both biodegradable and non-biodegradable peptide-loaded implants are already developed for the long-term treatment of patients, thereby reducing the frequency of drug administration. To further improve peptide formulation, extending the scope of implant-based drug delivery systems towards other polymers and processing techniques is highly interesting. OBJECTIVE: In this study, as a proof-of-principle, the feasibility of hot-melt processing of a peptide active pharmaceutical ingredient was assessed by developing a non-biodegradable poly(ethylenevinyl acetate) (33% VA) implant loaded with 20% (w/w) buserelin acetate. METHODS: Cross-sectional implant characterization was performed by Raman microscopy. The stability of buserelin acetate in the polymeric matrix was evaluated for 3 months under ICH stability conditions and the quantity as well as the degradation products analyzed using LC-UV methods. An in vitro dissolution study was performed as well and buserelin acetate and its degradants analyzed using the same chromatographic methods. RESULTS: No significant quantities of buserelin acetate-related degradation products were formed during the hot-melt preparation as well as during the stability study. Together with the consistent buserelin acetate assay values over time, chemical peptide stability was thus demonstrated. The in vitro buserelin acetate release from the implant was found to be diffusion-controlled after an initial burst release, with stable release profiles in the stability study, demonstrating the functional stability of the peptide implant. CONCLUSION: These results indicate the feasibility of preparing non-biodegradable peptide-loaded implants using the hot-melt production method and may act as a proof of principle concept for further innovation in peptide medicinal formulations.


Assuntos
Busserrelina/química , Portadores de Fármacos/química , Implantes de Medicamento/química , Polivinil/química , Busserrelina/administração & dosagem , Composição de Medicamentos/métodos , Implantes de Medicamento/administração & dosagem , Liberação Controlada de Fármacos , Estabilidade de Medicamentos , Estudos de Viabilidade , Temperatura Alta , Estudo de Prova de Conceito , Solubilidade
20.
Undersea Hyperb Med ; 46(1): 55-61, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31154685

RESUMO

Introduction: Eustachian tube dysfunction (ETD) and middle ear barotrauma (MEB) are the most common adverse effects of hyperbaric oxygen (HBO2) treatments. Patients practice equalization maneuvers to prevent ETD and MEB prior to hyperbaric exposure. Some patients are still unable to equalize middle ear pressure. This ETD results in undesirable consequences, including barotrauma, treatment with medications or surgical myringotomy with tube placement and interruption of HBO2. When additional medications and myringotomy are employed, they are associated with additional complications. Methods: A device known as the Ear Popper® has been reported to reduce complications from serous otitis media and reduce the need for surgical interventions (myringotomy). Patients unable to equalize middle ear pressure during initial compression in the hyperbaric chamber were allowed to use the device for rescue. All hyperbaric treatments were compressed using a United States Navy TT9, or a 45-fsw hyperbaric treatment schedule. Patients with persistent ETD and the inability to equalize middle ear pressure were given the Ear Popper upon consideration of terminating their treatment. Results: The Ear Popper allowed all patients to successfully equalize middle ear pressure and complete their treatments. Conclusion: This study substantiates the use of this device to assist in allowing pressurization of the middle ear space in patients otherwise unable to achieve equalization of middle ear pressure during HBO2 treatment in a multiplace chamber.


Assuntos
Barotrauma/prevenção & controle , Otopatias/prevenção & controle , Tuba Auditiva , Oxigenação Hiperbárica/efeitos adversos , Estudo de Prova de Conceito , Terapia de Salvação/instrumentação , Adulto , Idoso , Idoso de 80 Anos ou mais , Pressão Atmosférica , Deglutição , Desenho de Equipamento , Tuba Auditiva/fisiologia , Feminino , História do Século XIX , Humanos , Masculino , Pessoa de Meia-Idade , Otolaringologia/história , Terapia de Salvação/métodos
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