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1.
Diagnostics (Basel) ; 13(21)2023 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-37958262

RESUMO

The automated urine reagent strip test is a cost-effective tool for detecting albuminuria in patients. However, prior research has not investigated how urinary tract infections (UTIs) affect the test's accuracy. Therefore, this study aims to assess the impact of UTIs on albuminuria diagnosis using both the biochemical quantitative method and the test strip method of the Fully Automatic Urine Chemistry Analyzer, UC-3500 (Sysmex, Kobe, Japan). From March to December 2019, we prospectively collected midstream urine from adult female UTI patients before and after one week of cephalexin treatment. The urine samples were subjected to culture, routine urinalysis, and albuminuria diagnosis using the biochemical quantitative method and UC-3500. Albuminuria was defined as a urine albumin to creatinine ratio (UACR) ≥ 30 mg/g in the biochemical quantitative method. The results were compared between the two methods. Among fifty-four female patients (average age: 50.5 ± 4.4 years) with UTIs, 24 (44.44%) had transient albuminuria. The quantitative UACR significantly decreased after one week of antibiotic treatment (median: 53 mg/g to 9 mg/g; median difference: -0.54, p < 0.0001). UC-3500 exhibited a higher false positive rate for diagnosing albuminuria during UTIs (42%) compared to after treatment (19%). Its agreement with the biochemical quantitative method was moderate during UTI (κ = 0.49, 95% confidence interval [CI]: 0.24-0.73) and good after treatment (κ = 0.65, 95% CI: 0.45-0.86). UC-3500's accuracy in diagnosing albuminuria is influenced by UTIs, leading to either transient albuminuria or a false positive reaction of the test strip. UTI should be excluded or treated before its application in albuminuria screening.

2.
Anal Chim Acta ; 1275: 341593, 2023 Sep 22.
Artigo em Inglês | MEDLINE | ID: mdl-37524469

RESUMO

Whole-cell biosensors have demonstrated promising capabilities in detecting target molecules. However, their limited selectivity and precision can be attributed to the broad substrate tolerance of natural proteins. In this study, we aim to enhance the performance of whole-cell biosensors by incorporating of logic AND gates. Specifically, we utilize the HrpR/S system, a widely employed hetero-regulation module from Pseudomonas syringae in synthetic biology, to construct an orthogonal AND gate in Escherichia coli. To accomplish this, we compare the HrpR/S system with self-associating split fluorescent proteins using the Spy Tag/Spy Catcher system. Our objective is to selectively activate a reporter gene in the presence of both IPTG and Hg(II) ions. Through systematic genetic engineering and evaluation of various biological parts under diverse working conditions, our research demonstrates the utility of self-associating split fluorescent proteins in developing high-performance whole-cell biosensors. This approach offers advantages such as engineering simplicity, reduced basal activity, and improved selectivity. Furthermore, the comparison with the HrpR/S system serves as a valuable control model, providing insights into the relative advantages and limitations of each approach. These findings present a systematic and adaptable strategy to overcome the substrate tolerance challenge faced by whole-cell biosensors.

3.
Opt Express ; 30(26): 47004-47016, 2022 Dec 19.
Artigo em Inglês | MEDLINE | ID: mdl-36558638

RESUMO

We investigate the photonic topological phases in Tellegen metamaterials characterized by the antisymmetric magnetoelectric tensors with real-valued quantities. The underlying medium is considered a photonic analogue of the topological semimetal featured with a displaced Weyl cone in the frequency-wave vector space. As the 'spin'-degenerate condition is satisfied, the photonic system consists of two hybrid modes that are completely decoupled. By introducing the pseudospin states as the basis for the hybrid modes, the photonic system is described by two subsystems in terms of the spin-orbit Hamiltonians with spin 1, which result in nonzero spin Chern numbers that determine the topological properties. Surface modes at the interface between two Tellegen metamaterials with opposite sign of the magnetoelectric parameter exist at their common gap in the wave vector space, which are analytically formulated by algebraic equations. In particular, two types of surface modes are tangent to or wrapping around the Weyl cones, which form a pair of bended and a pair of twisted surface sheets. At the Weyl frequency, the surface modes contain a typical and two open Fermi arc-like states that concatenate to yield an infinite straight line. Topological features of the Tellegen metamaterials are further illustrated with the robust transport of surface modes at an irregular boundary.

4.
NPJ Digit Med ; 5(1): 166, 2022 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-36323795

RESUMO

Diabetic kidney disease is the leading cause of end-stage kidney disease worldwide; however, the integration of high-dimensional trans-omics data to predict this diabetic complication is rare. We develop artificial intelligence (AI)-assisted models using machine learning algorithms to identify a biomarker signature that predisposes high risk patients with diabetes mellitus (DM) to diabetic kidney disease based on clinical information, untargeted metabolomics, targeted lipidomics and genome-wide single nucleotide polymorphism (SNP) datasets. This involves 618 individuals who are split into training and testing cohorts of 557 and 61 subjects, respectively. Three models are developed. In model 1, the top 20 features selected by AI give an accuracy rate of 0.83 and an area under curve (AUC) of 0.89 when differentiating DM and non-DM individuals. In model 2, among DM patients, a biomarker signature of 10 AI-selected features gives an accuracy rate of 0.70 and an AUC of 0.76 when identifying subjects at high risk of renal impairment. In model 3, among non-DM patients, a biomarker signature of 25 AI-selected features gives an accuracy rate of 0.82 and an AUC of 0.76 when pinpointing subjects at high risk of chronic kidney disease. In addition, the performance of the three models is rigorously verified using an independent validation cohort. Intriguingly, analysis of the protein-protein interaction network of the genes containing the identified SNPs (RPTOR, CLPTM1L, ALDH1L1, LY6D, PCDH9, B3GNTL1, CDS1, ADCYAP and FAM53A) reveals that, at the molecular level, there seems to be interconnected factors that have an effect on the progression of renal impairment among DM patients. In conclusion, our findings reveal the potential of employing machine learning algorithms to augment traditional methods and our findings suggest what molecular mechanisms may underlie the complex interaction between DM and chronic kidney disease. Moreover, the development of our AI-assisted models will improve precision when diagnosing renal impairment in predisposed patients, both DM and non-DM. Finally, a large prospective cohort study is needed to validate the clinical utility and mechanistic implications of these biomarker signatures.

5.
Sci Rep ; 12(1): 18847, 2022 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-36344624

RESUMO

We investigate the photonic topological phases in pseudochiral metamaterials characterized by the magnetoelectric tensors with symmetric off-diagonal chirality components. The underlying medium is considered a photonic analogue of the type-II Weyl semimetal featured with two pairs of tilted Weyl cones in the frequency-wave vector space. As the 'spin'-degenerate condition is satisfied, the photonic system consists of two hybrid modes that are completely decoupled. By introducing the pseudospin states as the basis for the hybrid modes, the photonic system is described by two subsystems in terms of the spin-orbit Hamiltonians with spin 1, which result in nonzero spin Chern numbers that determine the topological properties. Surface modes at the interface between vacuum and the pseudochiral metamaterial exist in their common gap in the wave vector space, which are analytically formulated by algebraic equations. In particular, the surface modes are tangent to both the vacuum light cone and the Weyl cones, which form two pairs of crossing surface sheets that are symmetric about the transverse axes. At the Weyl frequency, the surface modes that connect the Weyl points form four Fermi arc-like states as line segments. Topological features of the pseudochiral metamaterials are further illustrated with the robust transport of surface modes at an irregular boundary.

6.
Biochem Pharmacol ; 206: 115327, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36330949

RESUMO

Triple-negative breast cancers (TNBCs) are difficult to cure and currently lack of effective treatment strategies. Cancer stem cells (CSCs) are highly associated with the poor clinical outcome of TNBCs. Thoc1 is a core component of the THO complex (THOC) that regulates the elongation, processing and nuclear export of mRNA. The function of thoc1 in TNBC and whether Thoc1 serves as a drug target are poorly understood. In this study, we demonstrated that thoc1 expression is elevated in TNBC cell lines and human TNBC patient tissues. Knockdown of thoc1 decreased cancer stem cell populations, reduced mammosphere formation, impaired THOC function, and downregulated the expression of stemness-related proteins. Moreover, the thoc1-knockdown 4T1 cells showed less lung metastasis in an orthotopic breast cancer mouse model. Overexpression of Thoc1 promoted TNBC malignancy and the mRNA export of stemness-related genes. Furthermore, treatment of TNBC cells with the natural compound andrographolide reduced the expression of Thoc1 expression, impaired homeostasis of THOC, suppressed CSC properties, and delayed tumor growth in a 4T1-implanted orthotopic mouse model. Andrographolide also reduced the activity of NF-κB, an upstream transcriptional regulator of Thoc1. Notably, thoc1 overexpression attenuates andrographolide-suppressed cellular proliferation. Altogether, our results demonstrate that THOC1 promotes cancer stem cell characteristics of TNBC, and andrographolide is a potential natural compound for eliminating CSCs of TNBCs by downregulating the NF-κB-thoc1 axis.


Assuntos
Neoplasias de Mama Triplo Negativas , Animais , Humanos , Camundongos , Linhagem Celular Tumoral , Proliferação de Células , Proteínas de Ligação a DNA/metabolismo , Células-Tronco Neoplásicas , NF-kappa B/metabolismo , RNA Mensageiro/metabolismo , Proteínas de Ligação a RNA/metabolismo , Neoplasias de Mama Triplo Negativas/metabolismo
8.
Biomedicines ; 10(1)2022 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-35052795

RESUMO

An increased risk of cardiovascular events was identified in patients with peripheral artery disease (PAD). Clopidogrel is one of the most widely used antiplatelet medications. However, there are heterogeneous outcomes when clopidogrel is used to prevent cardiovascular events in PAD patients. Here, we use an artificial intelligence (AI)-assisted methodology to identify genetic factors potentially involved in the clopidogrel-resistant mechanism, which is currently unclear. Several discoveries can be pinpointed. Firstly, a high proportion (>50%) of clopidogrel resistance was found among diabetic PAD patients in Taiwan. Interestingly, our result suggests that platelet function test-guided antiplatelet therapy appears to reduce the post-interventional occurrence of major adverse cerebrovascular and cardiac events in diabetic PAD patients. Secondly, AI-assisted genome-wide association study of a single-nucleotide polymorphism (SNP) database identified a SNP signature composed of 20 SNPs, which are mapped into 9 protein-coding genes (SLC37A2, IQSEC1, WASHC3, PSD3, BTBD7, GLIS3, PRDM11, LRBA1, and CNR1). Finally, analysis of the protein connectivity map revealed that LRBA, GLIS3, BTBD7, IQSEC1, and PSD3 appear to form a protein interaction network. Intriguingly, the genetic factors seem to pinpoint a pathway related to endocytosis and recycling of P2Y12 receptor, which is the drug target of clopidogrel. Our findings reveal that a combination of AI-assisted discovery of SNP signatures and clinical parameters has the potential to develop an ethnic-specific precision medicine for antiplatelet therapy in diabetic PAD patients.

9.
Int J Mol Sci ; 22(21)2021 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-34768917

RESUMO

Aging is the major risk factor for cardiovascular disease, which is the leading cause of mortality worldwide among aging populations. Cisd2 is a prolongevity gene that mediates lifespan in mammals. Previously, our investigations revealed that a persistently high level of Cisd2 expression in mice is able to prevent age-associated cardiac dysfunction. This study was designed to apply a genetic approach that induces cardiac-specific Cisd2 overexpression (Cisd2 icOE) at a late-life stage, namely a time point immediately preceding the onset of old age, and evaluate the translational potential of this approach. Several discoveries are pinpointed. Firstly, Cisd2 is downregulated in the aging heart. This decrease in Cisd2 leads to cardiac dysfunction and impairs electromechanical performance. Intriguingly, Cisd2 icOE prevents an exacerbation of age-associated electromechanical dysfunction. Secondly, Cisd2 icOE ameliorates cardiac fibrosis and improves the integrity of the intercalated discs, thereby reversing various structural abnormalities. Finally, Cisd2 icOE reverses the transcriptomic profile of the aging heart, changing it from an older-age pattern to a younger pattern. Intriguingly, Cisd2 icOE modulates a number of aging-related pathways, namely the sirtuin signaling, autophagy, and senescence pathways, to bring about rejuvenation of the heart as it enters old age. Our findings highlight Cisd2 as a novel molecular target for developing therapies targeting cardiac aging.


Assuntos
Envelhecimento/genética , Proteínas Relacionadas à Autofagia/genética , Fibrose Endomiocárdica/genética , Coração/fisiologia , Longevidade/genética , Proteínas do Tecido Nervoso/genética , Rejuvenescimento/fisiologia , Animais , Autofagia/genética , Proteínas Relacionadas à Autofagia/biossíntese , Senescência Celular/genética , Fibrose Endomiocárdica/fisiopatologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Proteínas do Tecido Nervoso/biossíntese , Sirtuínas/metabolismo , Transcriptoma/genética
10.
Cells ; 10(9)2021 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-34572079

RESUMO

Heart failure (HF) is a global pandemic public health burden affecting one in five of the general population in their lifetime. For high-risk individuals, early detection and prediction of HF progression reduces hospitalizations, reduces mortality, improves the individual's quality of life, and reduces associated medical costs. In using an artificial intelligence (AI)-assisted genome-wide association study of a single nucleotide polymorphism (SNP) database from 117 asymptomatic high-risk individuals, we identified a SNP signature composed of 13 SNPs. These were annotated and mapped into six protein-coding genes (GAD2, APP, RASGEF1C, MACROD2, DMD, and DOCK1), a pseudogene (PGAM1P5), and various non-coding RNA genes (LINC01968, LINC00687, LOC105372209, LOC101928047, LOC105372208, and LOC105371356). The SNP signature was found to have a good performance when predicting HF progression, namely with an accuracy rate of 0.857 and an area under the curve of 0.912. Intriguingly, analysis of the protein connectivity map revealed that DMD, RASGEF1C, MACROD2, DOCK1, and PGAM1P5 appear to form a protein interaction network in the heart. This suggests that, together, they may contribute to the pathogenesis of HF. Our findings demonstrate that a combination of AI-assisted identifications of SNP signatures and clinical parameters are able to effectively identify asymptomatic high-risk subjects that are predisposed to HF.


Assuntos
Predisposição Genética para Doença , Insuficiência Cardíaca/genética , Polimorfismo de Nucleotídeo Único , Idoso , Inteligência Artificial , Feminino , Estudo de Associação Genômica Ampla , Fatores de Risco de Doenças Cardíacas , Insuficiência Cardíaca/diagnóstico , Humanos , Masculino , Pessoa de Meia-Idade
11.
ACS Sens ; 6(8): 2868-2874, 2021 08 27.
Artigo em Inglês | MEDLINE | ID: mdl-34156242

RESUMO

Droplet digital loop-mediated isothermal amplification (ddLAMP) is an important assay for pathogen detection due to its high accuracy, specificity, and ability to quantify nucleic acids. However, performing ddLAMP requires expensive instrumentation and the need for highly trained personnel with expertise in microfluidics. To make ddLAMP more accessible, a ddLAMP assay is developed, featuring significantly decreased operational difficulty and instrumentation requirements. The proposed assay consists of three simplified steps: (1) droplet generation step, in which a LAMP mixture can be emulsified just by manually pulling a syringe connected to a microfluidic device. In this step, for the first time, we verify that highly monodispersed droplets can be generated with unstable flow rates or pressures, allowing untrained personnel to operate the microfluidic device and perform ddLAMP assay; (2) heating step, in which the droplets are isothermally heated in a water bath, which can be found in most laboratories; and (3) result analysis step, in which the ddLAMP result can be determined using only a fluorescence microscopy and an open-source analyzing software. Throughout the process, no droplet microfluidic expertise or equipment is required. More importantly, the proposed system enables multiple samples to be processed simultaneously with a detection limit of 10 copies/µL. The test is simple and intuitive to operate in most laboratories for multi-sample detection, significantly enhancing the accessibility and detection throughput of the ddLAMP technique.


Assuntos
Microfluídica , Técnicas de Amplificação de Ácido Nucleico , Dispositivos Lab-On-A-Chip , Técnicas de Diagnóstico Molecular
12.
Int J Mol Sci ; 21(23)2020 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-33287440

RESUMO

The ageing of human populations has become a problem throughout the world. In this context, increasing the healthy lifespan of individuals has become an important target for medical research and governments. Cardiac disease remains the leading cause of morbidity and mortality in ageing populations and results in significant increases in healthcare costs. Although clinical and basic research have revealed many novel insights into the pathways that drive heart failure, the molecular mechanisms underlying cardiac ageing and age-related cardiac dysfunction are still not fully understood. In this review we summarize the most updated publications and discuss the central components that drive cardiac ageing. The following characters of mitochondria-related dysfunction have been identified during cardiac ageing: (a) disruption of the integrity of mitochondria-associated membrane (MAM) contact sites; (b) dysregulation of energy metabolism and dynamic flexibility; (c) dyshomeostasis of Ca2+ control; (d) disturbance to mitochondria-lysosomal crosstalk. Furthermore, Cisd2, a pro-longevity gene, is known to be mainly located in the endoplasmic reticulum (ER), mitochondria, and MAM. The expression level of Cisd2 decreases during cardiac ageing. Remarkably, a high level of Cisd2 delays cardiac ageing and ameliorates age-related cardiac dysfunction; this occurs by maintaining correct regulation of energy metabolism and allowing dynamic control of metabolic flexibility. Together, our previous studies and new evidence provided here highlight Cisd2 as a novel target for developing therapies to promote healthy ageing.


Assuntos
Cálcio/metabolismo , Homeostase , Mitocôndrias Cardíacas/metabolismo , Animais , Biomarcadores , Sinalização do Cálcio , Reprogramação Celular , Senescência Celular/genética , Metabolismo Energético , Humanos , Espaço Intracelular/metabolismo , Lisossomos/metabolismo , Proteínas de Membrana/deficiência , Proteínas de Membrana/metabolismo , Mitocôndrias/metabolismo , Membranas Mitocondriais/metabolismo , Miocárdio/metabolismo , Miocárdio/ultraestrutura , Transdução de Sinais
13.
Sci Rep ; 10(1): 8422, 2020 05 21.
Artigo em Inglês | MEDLINE | ID: mdl-32439945

RESUMO

Toll-like receptors (TLRs) play crucial roles in host immune defenses. Recently, TLR-mediated autophagy is reported to promote immune responses via increasing antigen processing and presentation in antigen presenting cells. The present study examined whether the synthetic TLR4 activator (CCL-34) could induce autophagy to promote innate and adaptive immunity. In addition, the potential of CCL-34 as an immune adjuvant in vivo was also investigated. Our data using RAW264.7 cells and bone marrow-derived macrophages showed that CCL-34 induced autophagy through a TLR4-NF-κB pathway. The autophagy-related molecules (Nrf2, p62 and Beclin 1) were activated in RAW264.7 cells and bone marrow-derived macrophages under CCL-34 treatment. CCL-34-stimulated macrophages exhibited significant antigen-processing activity and induced the proliferation of antigen-specific CD4+T cells as well as the production of activated T cell-related cytokines, IL-2 and IFN-γ. Furthermore, CCL-34 immunization in mice induced infiltration of monocytes in the peritoneal cavity and elevation of antigen-specific IgG in the serum. CCL-34 treatment in vivo did not cause toxicity based on serum biochemical profiles. Notably, the antigen-specific responses induced by CCL-34 were attenuated by the autophagy inhibitor, 3-methyladenine. In summary, we demonstrated CCL-34 can induce autophagy to promote antigen-specific immune responses and act as an efficient adjuvant.


Assuntos
Adjuvantes Imunológicos/farmacologia , Autofagia/imunologia , Glicolipídeos/farmacologia , Imunogenicidade da Vacina/imunologia , Serina/análogos & derivados , Receptor 4 Toll-Like/metabolismo , Adenina/análogos & derivados , Adenina/farmacologia , Animais , Proteína Beclina-1/metabolismo , Linfócitos T CD4-Positivos/imunologia , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Imunoglobulina G/sangue , Interferon gama/metabolismo , Interleucina-2/metabolismo , Macrófagos/imunologia , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Monócitos/imunologia , Fator 2 Relacionado a NF-E2/metabolismo , Células RAW 264.7 , Serina/farmacologia , Vacinas/imunologia
14.
ACS Appl Bio Mater ; 3(8): 5120-5125, 2020 Aug 17.
Artigo em Inglês | MEDLINE | ID: mdl-35021688

RESUMO

The neurotransmitter phenylethylamine (PEA) is highly susceptible to oxidation to produce phenylacetic acid (PA). The fact that PEA and PA are both metabolites of phenylalanine (Phe) in humans makes them important indicators in the diagnosis of phenylketonuria. In this work, three-color whole-cell biosensors were developed to simultaneously detect these analytes (Phe, PEA, and PA). The tyrosine-responsive promoter was used to control the production of green fluorescent protein signals in response to Phe levels. The FeaR regulon was first used to indicate the presence of PEA, whereas the Paa regulon was used for the detection of PA. The combination of three sensor strains together made it possible to semiquantify the three analytes according to unique color outputs without cross-interference. We sought to optimize various modular components (ribosomal binding sites and fluorescent proteins) to ensure the rapid generation of fluorescent signals. Finally, the biosensors were implemented within a microchannel device to reduce sample consumption in point-of-care assays.

15.
PLoS Biol ; 17(10): e3000508, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31593566

RESUMO

CDGSH iron-sulfur domain-containing protein 2 (Cisd2) is pivotal to mitochondrial integrity and intracellular Ca2+ homeostasis. In the heart of Cisd2 knockout mice, Cisd2 deficiency causes intercalated disc defects and leads to degeneration of the mitochondria and sarcomeres, thereby impairing its electromechanical functioning. Furthermore, Cisd2 deficiency disrupts Ca2+ homeostasis via dysregulation of sarco/endoplasmic reticulum Ca2+-ATPase (Serca2a) activity, resulting in an increased level of basal cytosolic Ca2+ and mitochondrial Ca2+ overload in cardiomyocytes. Most strikingly, in Cisd2 transgenic mice, a persistently high level of Cisd2 is sufficient to delay cardiac aging and attenuate age-related structural defects and functional decline. In addition, it results in a younger cardiac transcriptome pattern during old age. Our findings indicate that Cisd2 plays an essential role in cardiac aging and in the heart's electromechanical functioning. They highlight Cisd2 as a novel drug target when developing therapies to delay cardiac aging and ameliorate age-related cardiac dysfunction.


Assuntos
Senilidade Prematura/genética , Envelhecimento/fisiologia , Bloqueio Atrioventricular/genética , Proteínas Relacionadas à Autofagia/genética , Coração/fisiopatologia , Proteínas do Tecido Nervoso/genética , ATPases Transportadoras de Cálcio do Retículo Sarcoplasmático/genética , Senilidade Prematura/metabolismo , Senilidade Prematura/fisiopatologia , Animais , Bloqueio Atrioventricular/diagnóstico por imagem , Bloqueio Atrioventricular/metabolismo , Bloqueio Atrioventricular/fisiopatologia , Proteínas Relacionadas à Autofagia/deficiência , Cálcio/metabolismo , Eletrocardiografia , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Coração/fisiologia , Homeostase/fisiologia , Masculino , Camundongos , Camundongos Knockout , Mitocôndrias Cardíacas/genética , Mitocôndrias Cardíacas/metabolismo , Miócitos Cardíacos/citologia , Miócitos Cardíacos/fisiologia , Proteínas do Tecido Nervoso/deficiência , Sarcômeros/fisiologia , ATPases Transportadoras de Cálcio do Retículo Sarcoplasmático/metabolismo , Transcriptoma
16.
Biochem Pharmacol ; 163: 308-320, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30822403

RESUMO

Chronic myelogenous leukemia (CML) is clinically treated with imatinib, which inhibits the kinase activity of the Bcr-Abl oncoprotein. However, imatinib resistance remains a common clinical issue. Andrographolide, the major compound of the medicinal plant Andrographis paniculata, was reported to exhibit anticancer activity. In this study, we explored the therapeutic potential of andrographolide and its derivative, NCTU-322, against both imatinib-sensitive and imatinib-resistant human CML cell lines. Both andrographolide and NCTU-322 downregulated the Bcr-Abl oncoprotein in imatinib-resistant CML cells through an Hsp90-dependent mechanism similar to that observed in imatinib-sensitive CML cells. In addition, NCTU-322 had stronger effects than andrographolide on downregulation of Bcr-Abl oncoprotein, induction of Hsp90 cleavage and cytotoxicity of CML cells. Notably, andrographolide and NCTU-322 could induce differentiation, mitotic arrest and apoptosis of both imatinib-sensitive and imatinib-resistant CML cells. Finally, the anticancer activity of NCTU-322 against imatinib-resistant CML cells was demonstrated in vivo. In summary, our data demonstrated that andrographolide and NCTU-322 inhibit Bcr-abl function via a mechanism different from that of imatinib, and they induced multiple anticancer effects in both imatinib-sensitive and resistant CML cells. Our findings demonstrate that andrographolide and NCTU-322 are potential therapeutic agents again CML.


Assuntos
Antineoplásicos/farmacologia , Diterpenos/farmacologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Genes abl/fisiologia , Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Antineoplásicos/química , Apoptose/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Diterpenos/química , Resistencia a Medicamentos Antineoplásicos , Genes abl/genética , Humanos , Mesilato de Imatinib/farmacologia , Leucócitos Mononucleares/efeitos dos fármacos , Estrutura Molecular
17.
Sci Rep ; 8(1): 17319, 2018 11 23.
Artigo em Inglês | MEDLINE | ID: mdl-30470789

RESUMO

Lateral flow immunoassays (LFIAs) have wide application in point-of-care testing, particularly in resource-poor settings. To achieve signal amplification in a gold nanoparticle-based lateral flow assay without an additional procedure or the need for complex fabrication, a new and simple method was developed for using a "stacking pad" configuration that adds an additional membrane between the conjugation pad and test pad to the conventional AuNP-based LFIA format. This design helps to accumulate the antibody and antigen on the stacking pad, hence extending the antigen/antibody binding interactions to enhance the test's detection sensitivity. With the enhanced lateral flow assay, as low as 1 ng/mL of Protein A and 15.5 ng/mL of C-reactive protein can be visualized with the naked eye. We also successfully applied the stacking pad system in the analysis of C-reactive protein in human serum and synovial fluid samples. These results suggest that this stacking pad LFIA can provide sensitive and on-site prognosis for detection in synovial fluid and serum samples in resource-limited settings.


Assuntos
Proteína C-Reativa/análise , Ouro/química , Imunoensaio/instrumentação , Imunoensaio/métodos , Nanopartículas Metálicas/química , Proteína Estafilocócica A/análise , Líquido Sinovial/metabolismo , Humanos
18.
Int J Syst Evol Microbiol ; 59(Pt 10): 2629-32, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19625439

RESUMO

A non-pigmented, Gram-negative, aerobic, rod-shaped bacterium, strain GR13(T), was isolated using nutrient agar from a water sample from a pond used for the culture of soft-shell turtles (Trionyx sinensis), Pingtung County, Taiwan. 16S rRNA gene sequence studies indicated that the novel strain formed a monophyletic branch at the periphery of the evolutionary radiation occupied by the genus Vogesella. Its closest neighbours were Vogesella indigofera ATCC 19706(T) and Vogesella perlucida DS-28(T) (both with 97.4 % gene sequence similarity). The novel isolate could be distinguished from these species by several phenotypic characteristics. The predominant fatty acids were summed feature 3 (C(16 : 1)omega7c and/or C(15 : 0) iso 2-OH; 60 %), C(16 : 0) (13.6 %) and C(18 : 1)omega7c (12.5 %). The DNA G+C content of strain GR13(T) was 63 mol%. The DNA-DNA hybridization values for the novel strain with V. indigofera and V. perlucida were <25 %. On the basis of the 16S rRNA gene sequence analysis and the chemotaxonomic and physiological data, it is concluded that strain GR13(T) represents a novel species in the genus Vogesella, for which the name Vogesella lacus sp. nov. is proposed. The type strain is GR13(T) (=BCRC 17836(T)=LMG 24504(T)).


Assuntos
Água Doce/microbiologia , Neisseriaceae/isolamento & purificação , Tartarugas , Animais , DNA Bacteriano/genética , DNA Ribossômico/genética , Ácidos Graxos/química , Ácidos Graxos/metabolismo , Dados de Sequência Molecular , Neisseriaceae/química , Neisseriaceae/classificação , Neisseriaceae/genética , Filogenia , RNA Ribossômico 16S/genética
19.
Int J Syst Evol Microbiol ; 58(Pt 12): 2677-81, 2008 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19060039

RESUMO

A transparent, non-pigmented, Gram-negative, rod-shaped bacterium, designated strain DS-28(T), was isolated from water samples collected from a spring located in Tainan County, Taiwan. 16S rRNA gene sequence analysis indicated that the novel strain formed a monophyletic branch at the periphery of the evolutionary radiation occupied by the genus Vogesella; the only sole close neighbour of the novel strain was Vogesella indigofera ATCC 19706(T) (97.4 % 16S rRNA gene sequence similarity). The isolate was distinguished from V. indigofera on the basis of genotypic data, several phenotypic properties and an inability to produce characteristic blue-pigmented colonies on peptone agar. The fatty acid profile was slightly different from that reported for V. indigofera ATCC 19706(T). It was evident from the genotypic and phenotypic data that strain DS-28(T) represents a novel species of the genus Vogesella, for which the name Vogesella perlucida sp. nov. is proposed. The type strain is DS-28(T) (=BCRC 17730(T)=LMG 24214(T)).


Assuntos
Água Doce/microbiologia , Neisseriaceae/classificação , Neisseriaceae/fisiologia , Ácidos Graxos/química , Genótipo , Dados de Sequência Molecular , Neisseriaceae/genética , Neisseriaceae/isolamento & purificação , Fenótipo , Filogenia , RNA Ribossômico 16S/genética , Especificidade da Espécie , Taiwan
20.
Int J Syst Evol Microbiol ; 58(Pt 4): 761-6, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18398166

RESUMO

A novel bacterial strain, designated a4T, isolated from a sea anemone (Aiptasia pulchella) in Taiwan, was characterized using a polyphasic taxonomic approach. Strain a4T was aerobic, Gram-negative, pale-yellow-pigmented and rod-shaped. It grew optimally at 30-35 degrees C, in the presence of 3-4 % (w/v) NaCl and at pH 8.0. Phylogenetic analyses based on 16S rRNA gene sequences showed that the strain belonged to the genus Tenacibaculum (family Flavobacteriaceae, phylum Bacteroidetes). The closest neighbours were Tenacibaculum lutimaris TF-26T (97.6 % similarity) and Tenacibaculum aestuarii SMK-4T (97.7 % similarity). The novel isolate could be distinguished from all Tenacibaculum species by several phenotypic characteristics. The major fatty acids were summed feature 3 (comprising C16 : 1 omega 7c and/or iso-C15 : 0 2-OH, 19.6 %), iso-C15 : 0 (12.9 %), iso-C16 : 0 3-OH (10.2 %), iso-C17 : 0 3-OH (9.9 %) and iso-C15 : 1 (9.5 %). The DNA G+C content was 35.0 mol%. Hence, genotypic and phenotypic data demonstrate that strain a4(T) should be classified as a representative of a novel species in the genus Tenacibaculum, for which the name Tenacibaculum aiptasiae sp. nov. is proposed. The type strain is a4T (=BCRC 17655T =LMG 24004T).


Assuntos
Flavobacteriaceae/genética , Flavobacteriaceae/isolamento & purificação , Anêmonas-do-Mar/microbiologia , Animais , Composição de Bases , DNA Bacteriano/química , DNA Bacteriano/genética , Ácidos Graxos/análise , Flavobacteriaceae/química , Flavobacteriaceae/classificação , Genes Bacterianos , Dados de Sequência Molecular , Fenótipo , Filogenia , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Especificidade da Espécie , Terminologia como Assunto
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