Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 27.602
Filtrar
1.
Arch Endocrinol Metab ; 63(4): 394-401, 2019 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-31365627

RESUMO

OBJECTIVE: To measure type 1 serum amino-terminal propeptide procollagen (P1NP) and type 1 cross-linked C-terminal telopeptide collagen (CTX) before parathyroidectomy (PTX) in PHPT patients, correlating these measurements with bone mineral density (BMD) changes. SUBJECTS AND METHODS: 31 primary hyperparathyroidism (HPTP) were followed from diagnosis up to 12-18 months after surgery. Serum levels of calcium, parathyroid hormone (PTH) vitamin D, CTX, P1NP, and BMD were measured before and 1 year after surgery. RESULTS: One year after PTX, the mean BMD increased by 8.6%, 5.5%, 5.5%, and 2.2% in the lumbar spine, femoral neck (FN), total hip (TH), and distal third of the nondominant radius (R33%), respectively. There was a significant correlation between BMD change 1 year after the PTX and CTX (L1-L4: r = 0.614, p < 0.0003; FN: r = 0.497, p < 0.0051; TH: r = 0.595, p < 0.0005; R33%: r = 0.364, p < 0.043) and P1NP (L1-L4: r = 0,687, p < 0,0001; FN: r = 0,533, p < 0,0024; TH: r = 0,642, p < 0,0001; R33%: r = 0,467, p < 0,0079) preoperative levels. The increase in 25(OH)D levels has no correlation with BMD increase (r = -0.135; p = 0.4816). On linear regression, a minimum preoperative CTX value of 0.331 ng/mL or P1NP of 37.9 ng/mL was associated with a minimum 4% increase in L1-L4 BMD. In TH, minimum preoperative values of 0.684 ng/mL for CTX and 76.0 ng/mL for P1NP were associated with a ≥ 4% increase in BMD. CONCLUSION: PHPT patients presented a significant correlation between preoperative levels of turnover markers and BMD improvement 1 year after PTX.


Assuntos
Densidade Óssea , Colágeno Tipo I/metabolismo , Hiperparatireoidismo Primário/metabolismo , Paratireoidectomia/reabilitação , Fragmentos de Peptídeos/metabolismo , Peptídeos/metabolismo , Pró-Colágeno/metabolismo , Idoso , Biomarcadores/sangue , Cálcio/sangue , Feminino , Humanos , Hiperparatireoidismo Primário/cirurgia , Masculino , Pessoa de Meia-Idade , Hormônio Paratireóideo/sangue , Fragmentos de Peptídeos/sangue , Período Pós-Operatório , Valor Preditivo dos Testes , Período Pré-Operatório , Pró-Colágeno/sangue , Estudos Retrospectivos , Vitamina D/sangue
2.
Cell Physiol Biochem ; 53(2): 413-428, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31415717

RESUMO

BACKGROUND/AIMS: Amyloid plaques, generated during the progression of Alzheimer's disease, cause major neurological deficits due to substantial cell toxicity and death. The underlying cause of plaque generation stems from cleavage of the amyloid precursor protein (APP) by ß-secretase (BACE1). A resulting amyloid-ß (Aß) fragment forms aggregates to produce the main constituent of a plaque. METHODS: Phage display and biopanning techniques were used to identify a 12-mer peptide that had a natural affinity for the BACE1 enzyme. The peptide was translated from phage DNA and synthetically produced. The peptide, at concentrations of 1nM, 10nM and 100nM, was used to confirm binding by direct assay. Non-specific binding to BACE2, renin and cathepsin D was tested by direct binding assay. A BACE1 activity assay was used to determine the peptide effect on cleavage of an APP substrate. Treatment of SY5Y cells with the peptide was used to determine toxicity and prevention of Aß40 and Aß42 production. RESULTS: After identification and synthetic production, the peptide exhibited a strong affinity for BACE1 at nanomolar concentrations in the direct assay. In case of non-specific binding to homologous BACE2, renin and cathepsin D, the peptide showed minor binding but was nullified when in solution with BACE1. The peptide addition to a BACE1 activity assay was able to significantly reduce the amount of substrate cleavage. SY5Y cells, when treated with the peptide, did not show any detrimental morphological changes while being able to reduce the production of natural Aß40 and Aß42. Even under stressed conditions (H2O2 treatment) where the Aß production was higher, the peptide was still able to significantly reduce the effect of BACE1 while not effecting cell viability. CONCLUSION: The identified peptide exhibited strong binding to BACE1 in vitro and was able to reduce production of Aß, suggesting a favourable BACE1 inhibitor for future refining and characterisation.


Assuntos
Doença de Alzheimer/tratamento farmacológico , Secretases da Proteína Precursora do Amiloide/antagonistas & inibidores , Peptídeos beta-Amiloides/antagonistas & inibidores , Ácido Aspártico Endopeptidases/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Fragmentos de Peptídeos/antagonistas & inibidores , Peptídeos/farmacologia , Doença de Alzheimer/metabolismo , Secretases da Proteína Precursora do Amiloide/metabolismo , Peptídeos beta-Amiloides/metabolismo , Ácido Aspártico Endopeptidases/metabolismo , Linhagem Celular , Descoberta de Drogas , Inibidores Enzimáticos/metabolismo , Humanos , Fragmentos de Peptídeos/metabolismo , Peptídeos/metabolismo
3.
Chem Commun (Camb) ; 55(59): 8595-8598, 2019 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-31276123

RESUMO

The amino acid sequence plays an essential role in amyloid formation. Here, using the central core recognition module of the Aß peptide and its reverse sequence, we show that although both peptides assemble into ß-sheets, their morphologies, kinetics and cell toxicities display marked differences. In addition, the native peptide, but not the reverse one, shows notable affinity towards bilayer lipid model membranes that modulates the aggregation pathways to stabilize the oligomeric intermediate states and function as the toxic agent responsible for neuronal dysfunction.


Assuntos
Peptídeos beta-Amiloides/química , Peptídeos beta-Amiloides/metabolismo , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/metabolismo , Sequência de Aminoácidos , Peptídeos beta-Amiloides/toxicidade , Animais , Linhagem Celular Tumoral , Colesterol/química , Humanos , Cinética , Bicamadas Lipídicas/química , Bicamadas Lipídicas/metabolismo , Fragmentos de Peptídeos/toxicidade , Fosfatidilcolinas/química , Conformação Proteica em Folha beta , Multimerização Proteica , Ratos , Esfingomielinas/química
4.
Food Chem ; 299: 125051, 2019 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-31284245

RESUMO

This study aims to exploit the molecular and cellular mechanisms concerning the functionality of dietary polyphenols (catechin, procyanidin B3, procyanidin C2, epigallocatechin and epigallocatechin gallate) in a nutritional context to prevent Celiac Disease (CD). In that sense, the interaction between the main CD bioactive peptide (32-mer peptide) and some polyphenols was fully characterized at the intestinal level under near physiological conditions by means of different spectroscopic techniques and dynamic simulations. Accordingly, it is proposed that the primarily polyphenol-binding sites on the 32-mer peptide correspond to leucine, tyrosine and phenylalanine containing domains being this interaction entropy-driven. Although procyanidin B3 and trimer C2 had a similar low-affinity constant at 310 K, both procyanidins were able to reduce the 32-mer peptide apical-to-basolateral translocation in in vitro simulated intestinal epithelial barrier thus prospecting the occurrence of additional and still unexplored regulatory mechanisms by which dietary polyphenols might modulate the transepithelial transport of CD bioactive peptides.


Assuntos
Alimentos , Glutens/química , Fragmentos de Peptídeos/química , Polifenóis/química , Análise Espectral , Glutens/metabolismo , Mucosa Intestinal/metabolismo , Fragmentos de Peptídeos/metabolismo , Ligação Proteica , Transporte Proteico
5.
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue ; 31(6): 680-683, 2019 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-31315722

RESUMO

OBJECTIVE: To explore the correlation between major inflammatory factors and septic shock in intensive care unit (ICU) patients, and to provide a basis for the diagnosis and treatment of septic shock. METHODS: The patients admitted to ICU of the Third People's Hospital of Datong from March 2017 to August 2018 were selected as the research objects. According to the diagnostic criteria of septic shock, the patients were divided into septic shock group and non-septic group. The basic information and inflammatory factors levels of the two groups, including age, gender, body mass index (BMI), course of disease, acute physiology and chronic health evaluation II (APACHE II), infection site and pathogenic; and C-reactive protein (CRP), procalcitonin (PCT), neutrophil lymphocyte ratio (NLR), N-terminal pro-B-type natriuretic peptide (NT-proBNP), tumor necrosis factor-α (TNF-α), γ-interferon (IFN-γ), interleukins (IL-1ß, IL-2, IL-6, IL-8) at 8 hours after diagnosis, were compared. Logistic regression was used to analyze the influencing factors of septic shock in ICU patients. RESULTS: A total of 154 ICU patients were selected, of whom 74 had septic shock. The APACHE II score of septic shock group was significantly higher than that of non-sepsis group (23.42±3.64 vs. 15.67±2.26, P < 0.05). There was no significant difference in other baseline data between the two groups. The levels of CRP, NT-proBNP, TNF-α, IFN-γ, PCT, IL-6, IL-8 in the septic shock group were significantly higher than those in the non-septic group [CRP (mg/L): 164.3±22.6 vs. 52.3±16.2, NT-proBNP (ng/L): 426.3±288.9 vs. 167.3±80.6, TNF-α (ng/L): 193.4±39.3 vs. 88.1±20.3, IFN-γ (ng/L): 133.3±52.0 vs. 97.0±56.1, PCT (ng/L): 27.6±10.2 vs. 7.3±4.1, IL-6 (ng/L): 83.0±17.6 vs. 20.9±6.4, IL-8 (ng/L): 445.8±34.0 vs. 84.0±25.7, all P < 0.05]. It was shown by Logistic regression analysis that CRP, NT-proBNP, TNF-α, PCT, IL-6 were independent risk factors for septic shock [CRP: odds ratio (OR) = 1.662, 95% confidence interval (95%CI) = 1.132-2.567; NT-proBNP: OR = 14.688, 95%CI = 3.580-20.238; TNF-α: OR = 1.093, 95%CI = 1.043-1.343; PCT: OR = 6.378, 95%CI = 4.556-12.243; IL-6: OR = 9.641, 95%CI = 2.242-13.786; all P < 0.05]. CONCLUSIONS: The levels of inflammatory factors CRP, NT-proBNP, TNF-α, PCT and IL-6 were significantly increased, which were important factors for early diagnosis of septic shock.


Assuntos
Choque Séptico/diagnóstico , Choque Séptico/metabolismo , APACHE , Proteína C-Reativa/metabolismo , Feminino , Humanos , Unidades de Terapia Intensiva , Interleucina-6/metabolismo , Masculino , Peptídeo Natriurético Encefálico/metabolismo , Fragmentos de Peptídeos/metabolismo , Pró-Calcitonina/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
6.
Eur J Med Chem ; 177: 291-301, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31158745

RESUMO

Based on our previous research on the fluorinated phenylquinoxaline scaffold, in this study, different positions of N,N-dimethyl amino group, and alkyl linkers with various lengths were introduced into this scaffold to regulate their lipophilicity and binding affinity to Tau. Four novel 99mTc/Re complexes with diethyl iminodiacetate chelator were synthesized and evaluated as Tau imaging tracers in the brain of Alzheimer's disease. Their specific binding to neurofibrillary tangles was verified by in vitro fluorescence staining and further confirmed by the results of immunofluorescence staining on the same brain sections from AD patient and Tg-tau mice. From in vitro binding assay using recombinant Tau aggregates, complex 4.2 with 6-N(CH3)2 and longer carbon chain (n = 4) displayed the highest affinity (Kd = 59.95 nM). [99mTc]4.2 was achieved by the ligand exchange reaction between dicarboxylic precursor and [99mTc(CO)3(H2O)3]+ intermediate with radiochemical yield over 45%. Ex vivo biodistribution studies on normal ICR mice revealed that [99mTc]4.2 exhibited moderate initial brain uptake (0.61% ID/g) and more structure optimizations are still required to improve the blood-brain barrier permeability.


Assuntos
Doença de Alzheimer/diagnóstico , Complexos de Coordenação/química , Corantes Fluorescentes/química , Compostos de Organotecnécio/química , Quinoxalinas/química , Proteínas tau/metabolismo , Idoso de 80 Anos ou mais , Peptídeos beta-Amiloides/metabolismo , Animais , Barreira Hematoencefálica/metabolismo , Encéfalo/metabolismo , Encéfalo/patologia , Quelantes/síntese química , Quelantes/química , Quelantes/metabolismo , Complexos de Coordenação/síntese química , Complexos de Coordenação/metabolismo , Feminino , Fluorescência , Corantes Fluorescentes/síntese química , Corantes Fluorescentes/metabolismo , Humanos , Iminoácidos/síntese química , Iminoácidos/química , Iminoácidos/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos ICR , Estrutura Molecular , Emaranhados Neurofibrilares/metabolismo , Emaranhados Neurofibrilares/patologia , Compostos de Organotecnécio/síntese química , Compostos de Organotecnécio/metabolismo , Fragmentos de Peptídeos/metabolismo , Quinoxalinas/síntese química , Quinoxalinas/metabolismo , Compostos Radiofarmacêuticos/síntese química , Compostos Radiofarmacêuticos/química , Compostos Radiofarmacêuticos/metabolismo , Rênio/química , Distribuição Tecidual
7.
Eur J Med Chem ; 177: 414-424, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31158754

RESUMO

Due to the role of butyrylcholinesterase (BChE) in acetylcholine hydrolysis in the late stages of the Alzheimer's disease (AD), inhibitors of butyrylcholinesterase (BChE) have been recently envisaged, besides acetylcholinesterase (AChE) inhibitors, as candidates for treating mild-to-moderate AD. Herein, synthesis and AChE/BChE inhibition activity of some twenty derivatives of 1,2,3,4,5,6-hexahydroazepino[4,3-b]indole (HHAI) is reported. Most of the newly synthesized HHAI derivatives achieved the inhibition of both ChE isoforms with IC50s in the micromolar range, with a structure-dependent selectivity toward BChE. Apparently, molecular volume and lipophilicity do increase selectivity toward BChE, and indeed the N2-(4-phenylbutyl) HHAI derivative 15d, which behaves as a mixed-type inhibitor, resulted the most potent (IC50 0.17 µM) and selective (>100-fold) inhibitor toward either horse serum and human BChE. Moreover, 15d inhibited in vitro self-induced aggregation of neurotoxic amyloid-ß (Aß) peptide and displayed neuroprotective effects in neuroblastoma SH-SY5Y cell line, significantly recovering (P < 0.001) cell viability when impaired by Aß1-42 and hydrogen peroxide insults. Overall, this study highlighted HHAI as useful and versatile scaffold for developing new small molecules targeting some enzymes and biochemical pathways involved in the pathogenesis of AD.


Assuntos
Azepinas/farmacologia , Inibidores da Colinesterase/farmacologia , Indóis/farmacologia , Fármacos Neuroprotetores/farmacologia , Peptídeos beta-Amiloides/metabolismo , Azepinas/síntese química , Azepinas/química , Azepinas/metabolismo , Butirilcolinesterase/química , Butirilcolinesterase/metabolismo , Domínio Catalítico , Linhagem Celular Tumoral , Inibidores da Colinesterase/síntese química , Inibidores da Colinesterase/química , Inibidores da Colinesterase/metabolismo , Relação Dose-Resposta a Droga , Desenho de Drogas , Depuradores de Radicais Livres/síntese química , Depuradores de Radicais Livres/química , Depuradores de Radicais Livres/metabolismo , Depuradores de Radicais Livres/farmacologia , Humanos , Indóis/síntese química , Indóis/química , Indóis/metabolismo , Simulação de Acoplamento Molecular , Estrutura Molecular , Fármacos Neuroprotetores/síntese química , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/metabolismo , Fragmentos de Peptídeos/metabolismo , Ligação Proteica/efeitos dos fármacos , Multimerização Proteica/efeitos dos fármacos , Relação Estrutura-Atividade
8.
Life Sci ; 231: 116537, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31176774

RESUMO

AIMS: Renal dysfunction has been reported in individuals with Down syndrome (DS); however, the causes and mechanisms involved remain unknown. Here, we present a proposal for how the triplication of the amyloid beta precursor protein (APP) and, mainly the amyloid ß peptide 1-42 (Aß42) can favor the development of renal abnormalities in DS. We evaluated the effects of vitamin D3 (VD3) supplementation on morphofunctional aspects and the repercussions on the presence and localization of Aß42, methylenetetrahydrofolate reductase (MTHFR), caspase-3 p12, and P-glycoprotein (Pgp) in the renal tissue of DS mouse model. MAIN METHODS: Twenty female mice (14-week-old) belonging to the B6EiC3Sn-Rb(12.Ts171665Dn)2Cje/CjeDnJ lineage were divided into four experimental groups (n = 5/group): common diet; trisomy (Ts) and wild-type (Wt); and high doses VD3, Ts(VD3), and Wt(VD3). All the groups were treated for 10 weeks. At 24 weeks, the protocol experimental was interrupted. The kidney was weighed, collected, and processed for immunochemical analysis for Aß42, Caspase-3 p12, MTHFR, and Pgp proteins. All data were analyzed statistically. KEY FINDINGS: Our results showed that VD3 promoted an increase in caspase-3 p12, MTHFR, and Pgp, and consequently contributed to reduced Aß42 in the renal tissue of a mouse model of DS. Furthermore, VD3 treatment affected the plasma creatinine and urea levels and contributed to the attenuation of the dilation of Bowman's space observed in trisomic mice. SIGNIFICANCE: Finally, the results showed that VD3 may activate specific mechanisms involved in reduced Aß42 and tissue repair in the kidneys of a mouse model for Down syndrome.


Assuntos
Peptídeos beta-Amiloides/metabolismo , Colecalciferol/farmacologia , Síndrome de Down/tratamento farmacológico , Síndrome de Down/metabolismo , Rim/efeitos dos fármacos , Fragmentos de Peptídeos/metabolismo , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Precursor de Proteína beta-Amiloide/metabolismo , Precursor de Proteína beta-Amiloide/fisiologia , Animais , Caspase 3/metabolismo , Suplementos Nutricionais , Modelos Animais de Doenças , Síndrome de Down/patologia , Feminino , Rim/metabolismo , Rim/patologia , Metilenotetra-Hidrofolato Redutase (NADPH2)/metabolismo , Camundongos , Camundongos Endogâmicos BALB C
9.
Life Sci ; 231: 116554, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31194992

RESUMO

AIMS: Several adipokines have been proven to improve the therapeutic efficacy of mesenchymal stromal cells (MSCs) when used to treat ischemic heart disease. Asprosin (ASP) is a newly-discovered adipokine. ASP might also predict the severity of coronary pathology. We investigated the role of ASP on MSCs and the effects of ASP-pretreated MSCs on myocardial infarction (MI). MAIN METHODS: MSCs were labelled with a lentivirus carrying green fluorescent protein (GFP). For in vivo study, after pretreatment with vehicle or ASP, MSCs were injected into infarcted hearts. Cardiac function and fibrosis were then evaluated 4 weeks after the induction of MI and survival of MSCs evaluated after 1 week. MSCs proliferation and migration were investigated after ASP treatment in vitro. MSCs apoptosis induced by hydrogen peroxide (H2O2) was assessed using flow cytometry. KEY FINDINGS: Compared to vehicle-pretreated MSCs, ASP-pretreated MSCs significantly improved the left ventricular ejection fraction (LVEF), and inhibited myocardial fibrosis 4 weeks after MI. ASP pretreatment may have promoted homing of transplanted MSCs. In vitro results showed that ASP had no significant effect on MSC proliferation and migration, but protected these cells from H2O2-induced apoptosis. Among 21 molecules associated with antioxidation and cell death, the antioxidant enzyme SOD2 was significantly upregulated by ASP. Furthermore, ASP treatment inhibited H2O2-induced ROS generation and apoptosis via the activated ERK1/2-SOD2 pathway. SIGNIFICANCE: This is the first evidence that ASP can regulate MSCs function and enhance MSCs therapy for ischemic heart disease. Furthermore, we demonstrate that ASP protects MSCs from oxidative stress-induced apoptosis via the ERK1/2-SOD2 pathway.


Assuntos
Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Células-Tronco Mesenquimais/efeitos dos fármacos , Proteínas dos Microfilamentos/metabolismo , Infarto do Miocárdio/terapia , Fragmentos de Peptídeos/metabolismo , Hormônios Peptídicos/metabolismo , Superóxido Dismutase/metabolismo , Animais , Apoptose/fisiologia , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Coração/fisiopatologia , Peróxido de Hidrogênio/farmacologia , Masculino , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/patologia , Camundongos , Camundongos Endogâmicos C57BL , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/patologia , Isquemia Miocárdica/metabolismo , Isquemia Miocárdica/patologia , Espécies Reativas de Oxigênio/metabolismo , Função Ventricular Esquerda
10.
Am J Orthod Dentofacial Orthop ; 155(5): 642-649, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31053279

RESUMO

INTRODUCTION: The purpose of this study was to investigate the effect of systemic delivery of Substance P (SP) on experimental tooth movement. METHODS: Forty-eight adult Sprague-Dawley rats were randomly divided into 2 groups and their maxillary first molars were mesially moved with the use of closed-coil springs. The experiment group received systemic injection of SP and the control group received phosphate-buffered saline solution. Transportation distances of first molars were measured. Hematoxylin and eosin staining, tartrate-resistant acid phosphatase staining, and immunohistochemistry staining were performed to evaluate alveolar bone remodeling. Then the interferon (IFN) γ and tumor necrosis factor (TNF) α concentrations in peripheral blood and local periodontal tissue were measured. Finally, the effects of SP on bone marrow-derived stem cell (BMSC) proliferation and migration were tested in vitro. RESULTS: Systemic delivery of SP significantly increased the distance of tooth movement and stimulated both osteoclast and osteoblast activities. The concentrations of IFN-γ and TNF-α increased in peripheral blood at early phases of the experiment and decreased in periodontal tissue at late phases. In vitro, the proliferation and migration of BMSCs were promoted by SP. CONCLUSIONS: Systemic delivery of SP can accelerate orthodontic tooth movement and promote alveolar bone remodeling potentially through immunomodulation and mobilizing endogenous mesenchymal stem cells.


Assuntos
Processo Alveolar/efeitos dos fármacos , Remodelação Óssea/efeitos dos fármacos , Substância P/farmacologia , Técnicas de Movimentação Dentária , Animais , Biomarcadores/metabolismo , Células da Medula Óssea/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Interferon gama/metabolismo , Maxila , Dente Molar , Osteoblastos/efeitos dos fármacos , Osteoclastos/efeitos dos fármacos , Fragmentos de Peptídeos/metabolismo , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Coloração e Rotulagem , Fator de Necrose Tumoral alfa/metabolismo
11.
Medicine (Baltimore) ; 98(20): e15247, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31096432

RESUMO

The aim of the current study was to investigate the expression of long non-coding RNA (lncRNA) antisense non-coding RNA in the INK4 locus (ANRIL) in allergic rhinitis (AR) patients, and to further explore the association of lncRNA ANRIL expression with AR risk, severity, and inflammation.In this case-control study, 96 AR patients and 96 non-atopic obstructive snoring patients who underwent adenoid surgery were consecutively recruited. Disease severity of AR patients was assessed via individual nasal symptom score (INSS) and total nasal symptom score (TNSS). Nasal mucosa samples were collected from AR patients and controls, then lncRNA ANRIL and inflammatory cytokine levels were assessed via quantitative polymerase chain reaction.LncRNA ANRIL expression was increased in AR patients (3.605 [1.763-4.981]) compared with controls (1.183 [0.438-2.985]), and it well distinguished AR patients from controls with an area under curve of 0.746 (95% CI: 0.679-0.814). Correlation analyses revealed that lncRNA ANRIL expression was positively associated with itching score and congestion score, while it was not associated with nasal rhinorrhea score or sneezing score. Besides, lncRNA ANRIL was also positively correlated with TNSS, tumor necrosis factor α, interleukin (IL)-4, IL-6, IL-13, and IL-17, while negatively associated with IL-10 and interferon-γ. And no association of lncRNA ANRIL expression with IL-1ß, IL-5, or IL-8 expression was discovered.LncRNA ANRIL expression correlates with increased AR risk, severity, and inflammation, implying that lncRNA ANRIL might be involved in the pathogenesis of AR.


Assuntos
Loci Gênicos/genética , Inflamação/metabolismo , Mucosa Nasal/metabolismo , RNA Longo não Codificante/genética , Rinite Alérgica/genética , Adolescente , Adulto , Estudos de Casos e Controles , Citocinas/metabolismo , Suscetibilidade a Doenças , Feminino , Humanos , Inflamação/patologia , Interleucina-1beta/metabolismo , Interleucina-5/metabolismo , Interleucina-8/metabolismo , Masculino , Mucosa Nasal/patologia , Fragmentos de Peptídeos/metabolismo , Rinite Alérgica/patologia , Rinite Alérgica/cirurgia , Índice de Gravidade de Doença , Fator de Necrose Tumoral alfa/metabolismo , Adulto Jovem
12.
Nat Commun ; 10(1): 2331, 2019 05 27.
Artigo em Inglês | MEDLINE | ID: mdl-31133680

RESUMO

Artificial nanoparticles accumulate a protein corona layer in biological fluids, which significantly influences their bioactivity. As nanosized obligate intracellular parasites, viruses share many biophysical properties with artificial nanoparticles in extracellular environments and here we show that respiratory syncytial virus (RSV) and herpes simplex virus type 1 (HSV-1) accumulate a rich and distinctive protein corona in different biological fluids. Moreover, we show that corona pre-coating differentially affects viral infectivity and immune cell activation. In addition, we demonstrate that viruses bind amyloidogenic peptides in their corona and catalyze amyloid formation via surface-assisted heterogeneous nucleation. Importantly, we show that HSV-1 catalyzes the aggregation of the amyloid ß-peptide (Aß42), a major constituent of amyloid plaques in Alzheimer's disease, in vitro and in animal models. Our results highlight the viral protein corona as an acquired structural layer that is critical for viral-host interactions and illustrate a mechanistic convergence between viral and amyloid pathologies.


Assuntos
Peptídeos beta-Amiloides/metabolismo , Herpesvirus Humano 1/patogenicidade , Interações Hospedeiro-Patógeno/imunologia , Fragmentos de Peptídeos/metabolismo , Coroa de Proteína/imunologia , Vírus Sincicial Respiratório Humano/patogenicidade , Doença de Alzheimer/imunologia , Doença de Alzheimer/patologia , Doença de Alzheimer/virologia , Animais , Líquido da Lavagem Broncoalveolar/virologia , Linhagem Celular Tumoral , Cercopithecus aethiops , Modelos Animais de Doenças , Feminino , Voluntários Saudáveis , Herpes Simples/sangue , Herpes Simples/imunologia , Herpes Simples/patologia , Herpesvirus Humano 1/imunologia , Humanos , Masculino , Camundongos , Camundongos Transgênicos , Agregados Proteicos/imunologia , Infecções por Vírus Respiratório Sincicial/imunologia , Infecções por Vírus Respiratório Sincicial/patologia , Infecções por Vírus Respiratório Sincicial/virologia , Vírus Sincicial Respiratório Humano/imunologia , Células Vero
13.
Food Chem ; 292: 217-226, 2019 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-31054668

RESUMO

Heat-stable peptidases released in refrigerated raw milk by psychrotrophic bacteria are responsible for UHT milk gelation. K-casein-derived caseinomacropeptides, identified by mass spectrometry, were constantly detected in gelled milk by capillary electrophoresis. Strains of Pseudomonas fluorescens, Ps. poae and Chryseobacterium joostei, selected among aprX-positive strains from raw milk, were incubated in milk up to 6 days at 4 °C before sterilization (98 °C/4 min). Samples were then stored at 25 or 40 °C, visually observed for gelation, and analysed for presence of caseinomacropeptides throughout 90 days of storage. Depending on cold pre-incubation time, caseinomacropeptides accumulated well before gelation onset in milk stored at 25 °C. Caseinomacropeptides were successively degraded, especially in milk stored at 40 °C, due to extensive proteolysis, and an abundant sediment developed instead of a gel. The caseinomacropeptides are here presented as an early indicator of UHT milk gelation and a mechanism explaining this phenomenon is proposed.


Assuntos
Caseínas/metabolismo , Géis/química , Leite/metabolismo , Pseudomonas fluorescens/fisiologia , Animais , Caseínas/análise , Caseínas/química , Cromatografia Líquida de Alta Pressão , Leite/microbiologia , Fragmentos de Peptídeos/análise , Fragmentos de Peptídeos/metabolismo , Peptídeo Hidrolases/metabolismo , Proteólise , Pseudomonas fluorescens/isolamento & purificação , Espectrometria de Massas em Tandem , Temperatura Ambiente
14.
Nanoscale ; 11(18): 9185-9193, 2019 May 09.
Artigo em Inglês | MEDLINE | ID: mdl-31038146

RESUMO

Recent advances in nanotechnology have developed a lot of opportunities for biological applications. In this work, multifunctional MoS2/AuNR nanocomposites with unique high NIR absorption were designed via combining MoS2 nanosheets and gold nanorods (AuNRs). The nanocomposites were synthesized through electrostatic self-assembly and showed high stability and good biocompatibility. Then they were used to modulate the aggregation of amyloid-ß peptides, destabilize mature fibrils under NIR irradiation, and eliminate Aß-induced ROS against neurotoxicity. The inhibition and destabilization effects were confirmed by Thioflavin T (ThT) fluorescence assay and transmission electron microscopy (TEM). Cell viability assay and ROS assay revealed that MoS2/AuNR nanocomposites could alleviate Aß-induced oxidative stress and cell toxicity. More importantly, both MoS2 nanosheets and AuNRs can be used as NIR photothermal agents, MoS2/AuNR nanocomposites have enhanced ability of disrupting Aß fibrils and improved cell viability by generating local heat under low power NIR irradiation. Our results provide new insights into the design of new multifunctional systems for the treatment of amyloid-related diseases.


Assuntos
Peptídeos beta-Amiloides/metabolismo , Raios Infravermelhos , Nanocompostos/química , Fragmentos de Peptídeos/metabolismo , Amiloide/química , Amiloide/toxicidade , Peptídeos beta-Amiloides/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Dissulfetos/química , Ouro/química , Humanos , Molibdênio/química , Nanocompostos/toxicidade , Nanotubos/química , Fragmentos de Peptídeos/química , Espécies Reativas de Oxigênio/metabolismo , Espectroscopia de Infravermelho com Transformada de Fourier
15.
Inorg Chem ; 58(11): 7488-7498, 2019 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-31083932

RESUMO

It was shown that His3 of human copper transporter 1 (hCtr1) prompts the ATCUN-like Cu(II) coordination for model peptides of the hCtr1 N-terminus. Its high Cu(II) affinity is a potential driving force for the transfer of Cu(II) from extracellular Cu(II) carriers to hCtr1. Having a sequence similar to that of hCtr1, hCtr2 has been proposed as another human copper transporter. However, the N-terminal domain of hCtr2 is much shorter than that of hCtr1, with different copper binding motifs at its N-terminus. Employing a model peptide of the hCtr2 N-terminus, MAMHF-am, we demonstrated that His4 provides a unique pattern of Cu(II) complexes, involving Met sulfurs in their Cu(II) coordination sphere. The affinity of Cu(II) for MAMHF-am is a few orders of magnitude lower than that reported for the hCtr1 model peptides at the extracellular pH of 7.4, suggesting a maximal complementary role of Cu(II) binding to hCtr2 in the import of copper from the extracellular space to the cytoplasm. On the other hand, the ability of the hCtr2 model peptide to capture Cu(II) from amino acids and short peptides (potential degradation products of proteins) at pH 5.0 and the known predominant lysosomal localization of hCtr2 support an important potential role of the Cu(II)-hCtr2 interaction in the recovery of copper from lysosomes.


Assuntos
Proteínas de Transporte de Cátions/química , Cobre/metabolismo , Espaço Extracelular/química , Lisossomos/química , Fragmentos de Peptídeos/metabolismo , Sequência de Aminoácidos , Espaço Extracelular/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Lisossomos/metabolismo , Modelos Moleculares , Fragmentos de Peptídeos/química , Ligação Proteica , Conformação Proteica
16.
Nat Commun ; 10(1): 2240, 2019 05 20.
Artigo em Inglês | MEDLINE | ID: mdl-31110178

RESUMO

The beta-site APP cleaving enzyme 1 (BACE1) is known primarily for its initial cleavage of the amyloid precursor protein (APP), which ultimately leads to the generation of Aß peptides. Here, we provide evidence that altered BACE1 levels and activity impact the degradation of Aß40 and Aß42 into a common Aß34 intermediate. Using human cerebrospinal fluid (CSF) samples from the Amsterdam Dementia Cohort, we show that Aß34 is elevated in individuals with mild cognitive impairment who later progressed to dementia. Furthermore, Aß34 levels correlate with the overall Aß clearance rates in amyloid positive individuals. Using CSF samples from the PREVENT-AD cohort (cognitively normal individuals at risk for Alzheimer's disease), we further demonstrate that the Aß34/Aß42 ratio, representing Aß degradation and cortical deposition, associates with pre-clinical markers of neurodegeneration. We propose that Aß34 represents a marker of amyloid clearance and may be helpful for the characterization of Aß turnover in clinical samples.


Assuntos
Doença de Alzheimer/patologia , Secretases da Proteína Precursora do Amiloide/metabolismo , Peptídeos beta-Amiloides/metabolismo , Ácido Aspártico Endopeptidases/metabolismo , Disfunção Cognitiva/patologia , Fragmentos de Peptídeos/metabolismo , Idoso , Doença de Alzheimer/líquido cefalorraquidiano , Secretases da Proteína Precursora do Amiloide/genética , Peptídeos beta-Amiloides/líquido cefalorraquidiano , Animais , Ácido Aspártico Endopeptidases/genética , Biomarcadores/líquido cefalorraquidiano , Biomarcadores/metabolismo , Encéfalo/patologia , Linhagem Celular Tumoral , Disfunção Cognitiva/líquido cefalorraquidiano , Estudos de Coortes , Progressão da Doença , Feminino , Humanos , Masculino , Camundongos , Camundongos Transgênicos , Pessoa de Meia-Idade , Fragmentos de Peptídeos/líquido cefalorraquidiano , Proteólise , Ratos , Ratos Sprague-Dawley
17.
Nat Methods ; 16(6): 509-518, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31133760

RESUMO

In mass-spectrometry-based proteomics, the identification and quantification of peptides and proteins heavily rely on sequence database searching or spectral library matching. The lack of accurate predictive models for fragment ion intensities impairs the realization of the full potential of these approaches. Here, we extended the ProteomeTools synthetic peptide library to 550,000 tryptic peptides and 21 million high-quality tandem mass spectra. We trained a deep neural network, termed Prosit, resulting in chromatographic retention time and fragment ion intensity predictions that exceed the quality of the experimental data. Integrating Prosit into database search pipelines led to more identifications at >10× lower false discovery rates. We show the general applicability of Prosit by predicting spectra for proteases other than trypsin, generating spectral libraries for data-independent acquisition and improving the analysis of metaproteomes. Prosit is integrated into ProteomicsDB, allowing search result re-scoring and custom spectral library generation for any organism on the basis of peptide sequence alone.


Assuntos
Aprendizado Profundo , Redes Neurais (Computação) , Fragmentos de Peptídeos/análise , Biblioteca de Peptídeos , Proteoma/análise , Software , Espectrometria de Massas em Tandem/métodos , Animais , Caenorhabditis elegans/metabolismo , Bases de Dados de Proteínas , Drosophila melanogaster/metabolismo , Células HEK293 , Humanos , Fragmentos de Peptídeos/metabolismo , Proteoma/metabolismo , Saccharomyces cerevisiae/metabolismo
18.
Nat Methods ; 16(6): 519-525, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31133761

RESUMO

Peptide fragmentation spectra are routinely predicted in the interpretation of mass-spectrometry-based proteomics data. However, the generation of fragment ions has not been understood well enough for scientists to estimate fragment ion intensities accurately. Here, we demonstrate that machine learning can predict peptide fragmentation patterns in mass spectrometers with accuracy within the uncertainty of measurement. Moreover, analysis of our models reveals that peptide fragmentation depends on long-range interactions within a peptide sequence. We illustrate the utility of our models by applying them to the analysis of both data-dependent and data-independent acquisition datasets. In the former case, we observe a q-value-dependent increase in the total number of peptide identifications. In the latter case, we confirm that the use of predicted tandem mass spectrometry spectra is nearly equivalent to the use of spectra from experimental libraries.


Assuntos
Biomarcadores/sangue , Análise de Dados , Fragmentos de Peptídeos/análise , Biblioteca de Peptídeos , Proteoma/análise , Software , Espectrometria de Massas em Tandem/métodos , Algoritmos , Sequência de Aminoácidos , Bases de Dados de Proteínas , Células HeLa , Humanos , Fragmentos de Peptídeos/metabolismo , Proteoma/metabolismo
19.
APMIS ; 127(7): 515-528, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31009118

RESUMO

The aim of this study was to examine the changes in hemostasis parameters in endocarditis and thromboembolic events in nonfatal methicillin-sensitive Staphylococcus aureus bacteremia (MS-SAB) - a topic not evaluated previously. In total, 155 patients were recruited and were categorized according to the presence of endocarditis or thromboembolic events with gender-age adjusted controls. Patients who deceased within 90 days or patients not chosen as controls were excluded. SAB management was supervised by an infectious disease specialist. Patients with endocarditis (N = 21), compared to controls (N = 21), presented lower antithrombin III at day 4 (p < 0.05), elevated antithrombin III at day 90 (p < 0.01), prolonged activated partial thromboplastin time at days 4 and 10 (p < 0.05), and enhanced thrombin-antithrombin complex at day 4 (p < 0.01). Thromboembolic events (N = 8), compared to controls (N = 34), significantly increased thrombin-antithrombin complex at day 4 (p < 0.05). In receiver operating characteristic analysis, the changes in these hemostasis parameters at day 4 predicted endocarditis and thromboembolic events (p < 0.05). No differences in hemoglobin, thrombocyte, prothrombin fragment, thrombin time, factor VIII, D-dimer or fibrinogen levels were observed between cases and controls. The results suggest that nonfatal MS-SAB patients present marginal hemostasis parameter changes that, however, may have predictability for endocarditis or thromboembolic events. Larger studies are needed to further assess the connection of hemostasis to complications in SAB.


Assuntos
Bacteriemia/complicações , Endocardite Bacteriana/etiologia , Hemostasia/fisiologia , Infecções Estafilocócicas/complicações , Tromboembolia/etiologia , Antitrombina III/metabolismo , Bacteriemia/metabolismo , Plaquetas/metabolismo , Plaquetas/fisiologia , Fator VIII/metabolismo , Feminino , Hemoglobinas/metabolismo , Humanos , Masculino , Meticilina/farmacologia , Pessoa de Meia-Idade , Fragmentos de Peptídeos/metabolismo , Peptídeo Hidrolases/metabolismo , Estudos Prospectivos , Infecções Estafilocócicas/metabolismo , Staphylococcus aureus/patogenicidade , Tempo de Trombina/métodos , Tromboembolia/metabolismo
20.
Philos Trans A Math Phys Eng Sci ; 377(2147): 20180422, 2019 Jun 17.
Artigo em Inglês | MEDLINE | ID: mdl-31030650

RESUMO

Structure-guided drug discovery emerged in the 1970s and 1980s, stimulated by the three-dimensional structures of protein targets that became available, mainly through X-ray crystal structure analysis, assisted by the development of synchrotron radiation sources. Structures of known drugs or inhibitors were used to guide the development of leads. The growth of high-throughput screening during the late 1980s and the early 1990s in the pharmaceutical industry of chemical libraries of hundreds of thousands of compounds of molecular weight of approximately 500 Da was impressive but still explored only a tiny fraction of the chemical space of the predicted 1040 drug-like compounds. The use of fragments with molecular weights less than 300 Da in drug discovery not only decreased the chemical space needing exploration but also increased promiscuity in binding targets. Here we discuss advances in X-ray fragment screening and the challenge of identifying sites where fragments not only bind but can be chemically elaborated while retaining their positions and binding modes. We first describe the analysis of fragment binding using conventional X-ray difference Fourier techniques, with Mycobacterium abscessus SAICAR synthetase (PurC) as an example. We observe that all fragments occupy positions predicted by computational hotspot mapping. We compare this with fragment screening at Diamond Synchrotron Light Source XChem facility using PanDDA software, which identifies many more fragment hits, only some of which bind to the predicted hotspots. Many low occupancy sites identified may not support elaboration to give adequate ligand affinity, although they will likely be useful in drug discovery as 'warm spots' for guiding elaboration of fragments bound at hotspots. We discuss implications of these observations for fragment screening at the synchrotron sources. This article is part of the theme issue 'Fifty years of synchrotron science: achievements and opportunities'.


Assuntos
Descoberta de Drogas/história , Síncrotrons/história , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Sítios de Ligação , Descoberta de Drogas/métodos , Descoberta de Drogas/tendências , Ensaios de Triagem em Larga Escala/história , Ensaios de Triagem em Larga Escala/métodos , Ensaios de Triagem em Larga Escala/tendências , História do Século XX , História do Século XXI , Humanos , Modelos Moleculares , Mycobacterium abscessus/efeitos dos fármacos , Mycobacterium abscessus/metabolismo , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/metabolismo , Peptídeo Sintases/química , Peptídeo Sintases/metabolismo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA