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1.
Nutrients ; 16(2)2024 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-38276557

RESUMO

Panax ginseng Meyer and Inula japonica Thunb. are well established in traditional medicine and are known for their therapeutic properties in managing a range of ailments such as diabetes, asthma, and cancer. Although P. ginseng and I. japonica can alleviate pulmonary fibrosis (PF), the anti-fibrosis effect on PF by the combination of two herbal medicines remains unexplored. Therefore, this study explores this combined effect. In conditions that were not cytotoxic, MRC-5 cells underwent treatment using the formula combining P. ginseng and I. japonica (ISE081), followed by stimulation with transforming growth factor (TGF)-ß1, to explore the fibroblast-to-myofibroblast transition (FMT). After harvesting the cells, mRNA levels and protein expressions associated with inflammation and FMT-related markers were determined to evaluate the antiinflammation activities and antifibrosis effect of ISE081. Additionally, the anti-migratory effects of ISE081 were validated through a wound-healing assay. ISE081 remarkably reduced the mRNA levels of interleukin (IL)-6, IL-8, α-smooth muscle actin (SMA), and TGF-ß1 in MRC-5 cells and suppressed the α-SMA and fibronectin expressions, respectively. Furthermore, ISE081 inhibited Smad2/3 phosphorylation and wound migration of MRC-5 cells. Under the same conditions, comparing those of ISE081, P. ginseng did not affect the expression of α-SMA, fibronectin, and Smad2/3 phosphorylation, whereas I. japonica significantly inhibited them but with cytotoxicity. The results indicate that the synergistic application of P. ginseng and I. japonica enhances the anti-fibrotic properties in pulmonary fibroblasts and concurrently diminishes toxicity. Therefore, ISE081 has the potential as a prevention and treatment herbal medicine for PF.


Assuntos
Inula , Panax , Fibrose Pulmonar , Humanos , Inula/metabolismo , Fibronectinas/genética , Fibronectinas/metabolismo , Panax/metabolismo , Fibrose , Fibrose Pulmonar/metabolismo , Fibroblastos , Fator de Crescimento Transformador beta1/metabolismo , RNA Mensageiro/metabolismo
2.
Sensors (Basel) ; 20(18)2020 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-32942708

RESUMO

Graphene oxide (GO)/peptide complexes as a promising disease biomarker analysis platform have been used to detect proteolytic activity by observing the turn-on signal of the quenched fluorescence upon the release of peptide fragments. However, the purification steps are often cumbersome during surface modification of nano-/micro-sized GO. In addition, it is still challenging to incorporate the specific peptides into GO with proper orientation using conventional immobilization methods based on pre-synthesized peptides. Here, we demonstrate a robust magnetic GO (MGO) fluorescence resonance energy transfer (FRET) platform based on in situ sequence-specific peptide synthesis of MGO. The magnetization of GO was achieved by co-precipitation of an iron precursor solution. Magnetic purification/isolation enabled efficient incorporation of amino-polyethylene glycol spacers and subsequent solid-phase peptide synthesis of MGO to ensure the oriented immobilization of the peptide, which was evaluated by mass spectrometry after photocleavage. The FRET peptide MGO responded to proteases such as trypsin, thrombin, and ß-secretase in a concentration-dependent manner. Particularly, ß-secretase, as an important Alzheimer's disease marker, was assayed down to 0.125 ng/mL. Overall, the MGO platform is applicable to the detection of other proteases by using various peptide substrates, with a potential to be used in an automated synthesis system operating in a high throughput configuration.


Assuntos
Transferência Ressonante de Energia de Fluorescência , Grafite , Peptídeo Hidrolases , Peptídeos/síntese química , Óxidos
3.
Biomacromolecules ; 19(12): 4534-4541, 2018 12 10.
Artigo em Inglês | MEDLINE | ID: mdl-30475587

RESUMO

Peptides are suitable candidates for templates in the fabrication of various metal nanoparticles (NPs) because of their metal-binding ability and templating effect, which impart physicochemical properties to the produced nanoparticles. Peptide-binding gold nanoparticles (AuNPs) show high catalytic activity that permits their application in oxidation or reduction reactions. Herein, we prepared morphology-controllable AuNPs stabilized by self-assembled tyrosine-rich peptides (YC7) by varying the pH and YC7 peptide/Au3+ concentration ratio in 2-( N-morpholino)ethanesulfonic acid (MES) buffer solution. The catalytic activities of the YC7 peptide-stabilized AuNPs (YC7@AuNPs) were tested for 4-nitrophenol (4-NP) reduction, and kinetic analysis was performed to calculate the apparent rate constants and activation energies. The relatively low activation energy of the YC7@AuNPs could be explained by the hypothesis that the tyrosine-moiety of YC7 enriches the electron density of Au metal.


Assuntos
Cinética , Nanopartículas Metálicas/química , Nitrofenóis/química , Peptídeos/química , Catálise , Ouro/química , Oxirredução , Tamanho da Partícula , Tirosina/química
4.
J Nanosci Nanotechnol ; 17(2): 1083-091, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29676553

RESUMO

This paper proposes Fiber-Optic Localized Surface Plasmon Resonance (FO LSPR) sensor combined with a micro fluidic channel, which enables continuous supply of fluid for bio-reaction. The proposed method prevents degradation of the sensing performance due to changes in measurement conditions. The feasibility of the FO LSPR sensor with a micro fluidic channel was demonstrated by computational fluid dynamics (CFD) simulation. Also, the proposed method was assessed by measuring the output intensity of the FO LSPR sensor at various refractive index solutions. Finally, a prostate-specific antigen (PSA) immunoassay was performed to evaluate the possibility of the fabricated sensor system as a biosensor.

5.
J Control Release ; 237: 177-184, 2016 09 10.
Artigo em Inglês | MEDLINE | ID: mdl-27349354

RESUMO

A disulfide-bridged cyclic RGD peptide, named iRGD (internalizing RGD, c(CRGDK/RGPD/EC)), is known to facilitate tumor targeting as well as tissue penetration. After the RGD motif-induced targeting on αv integrins expressed near tumor tissue, iRGD encounters proteolytic cleavage to expose the CendR motif that promotes penetration into cancer cells via the interaction with neuropilin-1. Based on these proteolytic cleavage and internalization mechanism, we designed an iRGD-based monolithic imaging probe that integrates multiple functions (cancer-specific targeting, internalization and fluorescence activation) within a small peptide framework. To provide the capability of activatable fluorescence signaling, we conjugated a fluorescent dye to the N-terminal of iRGD, which was linked to the internalizing sequence (CendR motif), and a quencher to the opposite C-terminal. It turned out that fluorescence activation of the dye/quencher-conjugated monolithic peptide probe requires dual (reductive and proteolytic) cleavages on both disulfide and amide bond of iRGD peptide. Furthermore, the cleavage of the iRGD peptide leading to fluorescence recovery was indeed operative depending on the tumor-related angiogenic receptors (αvß3 integrin and neuropilin-1) in vitro as well as in vivo. Compared to an 'always fluorescent' iRGD control probe without quencher conjugation, the dye/quencher-conjugated activatable monolithic peptide probe visualized tumor regions more precisely with lower background noise after intravenous injection, owing to the multifunctional responses specific to tumor microenvironment. All these results, along with minimal in vitro and in vivo toxicity profiles, suggest potential of the iRGD-based activatable monolithic peptide probe as a promising imaging agent for precise tumor diagnosis.


Assuntos
Corantes Fluorescentes/metabolismo , Integrina alfaVbeta3/metabolismo , Neoplasias/diagnóstico por imagem , Neuropilina-1/metabolismo , Oligopeptídeos/metabolismo , Imagem Óptica/métodos , Animais , Linhagem Celular Tumoral , Sistemas de Liberação de Medicamentos , Fluorescência , Corantes Fluorescentes/análise , Humanos , Camundongos , Neoplasias/metabolismo , Oligopeptídeos/análise
6.
Biosci Biotechnol Biochem ; 79(8): 1362-5, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25744337

RESUMO

The immuno-modulating activities of seaweed (Hizikia fusiforme) extracts on murine macrophage and splenocyte were studied in vitro. Polysaccharide (HFP) exhibited the potential macrophage stimulating effects than water extract (HFW) such as NO production and enhanced pro-inflammatory cytokines on the Raw 264.7 cells and splenocytes. From the mono-sugar composition, HFP-associated fucose based on HFP of H. fusiforme acts as immune modulator.


Assuntos
Fatores Imunológicos/administração & dosagem , Macrófagos/efeitos dos fármacos , Extratos Vegetais/administração & dosagem , Polissacarídeos/administração & dosagem , Animais , Camundongos , Phaeophyceae/química , Extratos Vegetais/química , Extratos Vegetais/imunologia , Polissacarídeos/química , Polissacarídeos/imunologia
7.
J Biomed Mater Res A ; 103(1): 365-70, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24665040

RESUMO

Astrocytes are known to serve as scaffolding cells that shape the brain. The physical properties of astrocytes, such as stiffness, are important for their scaffolding function. These properties may be altered in certain pathological conditions, such as in brain cancer. However, actual stiffness of astrocytes is not yet well understood. Here, we report that the astrocyte stiffness is positively correlated with the density of cytoskeletal proteins, such as actin filaments, microtubules, and intermediate filaments. The value of the stiffness of astrocytes as measured by atomic force microscopy (AFM) increases 38-fold in five-week-old rats compared to postnatal-day zero pups. Using multicolor confocal microscopy, we found that the complexity of cytoskeletal proteins, such as actin filaments, microtubules, and intermediate filaments, increase as the animal gets older. Our findings indicate that the change of stiffness positively correlates with the maturation of cytoskeletal proteins, and suggest that AFM can be useful as an analytical and diagnostic tool for neuroscience.


Assuntos
Astrócitos/citologia , Citoesqueleto/metabolismo , Nanotecnologia , Animais , Animais Recém-Nascidos , Imunofluorescência , Microscopia de Força Atômica , Ratos , Ratos Sprague-Dawley
8.
Sensors (Basel) ; 14(12): 22199-207, 2014 Nov 25.
Artigo em Inglês | MEDLINE | ID: mdl-25429407

RESUMO

The ability to measure pressure and force is essential in biomedical applications such as minimally invasive surgery (MIS) and palpation for detecting cancer cysts. Here, we report a force sensor for measuring a shear and normal force by combining an arrayed piezoelectric sensors layer with a precut glass top plate connected by four stress concentrating legs. We designed and fabricated a thin film piezoelectric force sensor and proposed an enhanced sensing tool to be used for analyzing gentle touches without the external voltage source used in FET sensors. Both the linear sensor response from 3 kPa to 30 kPa and the exact signal responses from the moving direction illustrate the strong feasibility of the described thin film miniaturized piezoelectric force sensor.


Assuntos
Biomimética/instrumentação , Manometria/instrumentação , Membranas Artificiais , Sistemas Microeletromecânicos/instrumentação , Tato , Transdutores de Pressão , Miniaturização , Pressão , Estresse Mecânico
9.
Phytother Res ; 28(12): 1893-6, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25042112

RESUMO

In the course of this experiment on the anti-inflammatory effect of ginsenosides, protopanaxdiol ginsenosides have shown inhibition activities in inflammatory responses: NF-κB, COX-2, and iNOS were induced by TNF-α. The responses of this experiment were evaluated by NF-κB-luciferase assay and RT-PCR experiment of COX-2 and iNOS genes. The NF-κB expressions were inhibited by ginsenosides Rd, Rg5 , Rz1 , and Rk1 in a dose-dependent manner. The IC50 values were 3.47, 0.61, 0.63, and 0.75 µM, respectively. Particularly, ginsenosides Rg5 , Rz1 , and Rk1 as converted ginsenosides from primary protopanaxdiol ginsenosidess significantly inhibited COX-2 and iNOS gene expression. These inhibition levels were similar to sulfasalazine as reference material.


Assuntos
Anti-Inflamatórios/farmacologia , Ciclo-Oxigenase 2/metabolismo , Ginsenosídeos/farmacologia , NF-kappa B/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Relação Dose-Resposta a Droga , Expressão Gênica , Células Hep G2 , Humanos , Concentração Inibidora 50 , Fator de Necrose Tumoral alfa/metabolismo
10.
Arch Pharm Res ; 37(4): 467-72, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24014307

RESUMO

A new neolignan, linderin A (1), together with six known lignans, (+)-xanthoxyol (2), pluviatilol (3), actiforin (4), (+)-syringaresinol (5), (+)-(7S,8R,8'R)-acuminatolide (6), and (+)-9'-O-trans-feruloyl-5,5'-dimethoxylariciresinol (7) were isolated from the stems of Lindera obtusiloba Blume (Lauraceae). Their chemical structures were elucidated by a combination of spectroscopic analysis and chemical reaction, and the absolute configuration of 1 was determined by Mosher's esterification. The effect of compounds 1-7 on tumor necrosis factor (TNF)-α, interleukin(IL)-6, and their inhibitory activity of histamine release were examined using human mast cells. Among the lignans tested, compounds 1, 3, 4, 6, and 7 inhibited release of histamine from mast cells. Especially, compounds 1 and 4 suppressed the gene expressions of pro-inflammatory cytokines, TNF-α, and IL-6 in human mast cells. Our findings suggest that the lignan constituents in L. obtusiloba may contribute to its anti-allergic inflammatory effects.


Assuntos
Antialérgicos/isolamento & purificação , Antialérgicos/farmacologia , Lignanas/isolamento & purificação , Lignanas/farmacologia , Lindera/química , Antialérgicos/química , Liberação de Histamina/efeitos dos fármacos , Humanos , Interleucina-6/metabolismo , Lignanas/química , Mastócitos/efeitos dos fármacos , Mastócitos/metabolismo , Extratos Vegetais/farmacologia , Caules de Planta/química , Relação Estrutura-Atividade , Fator de Necrose Tumoral alfa/metabolismo
11.
J Ethnopharmacol ; 150(3): 875-85, 2013 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-24140584

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Ganoderma lucidum (Fr.) Karst. (Ganodermataceae) is a mushroom which is used as a traditional remedy in the treatment of human diseases such as hepatitis, liver disorders, hypercholesterolemia, arthritis, bronchitis and tumorigenic diseases. This study targets the evaluation of hepatoprotective activity of ganodermanontriol, a sterol isolated from Ganoderma lucidum, and the investigation of its mechanism of action in Hepa1c1c7 and murine liver cells upon tert-butyl hydroperoxide (t-BHP)-induced inflammation. t-BHP was utilized to stimulate an anti-inflammatory reaction in the hepatic cell lines and murine hepatic tissue examined. Western blot and reverse transcription-quantitative polymerase chain reaction (RT-PCR) were used to estimate the expression of ganodermanontriol (GDT)-induced proteins, including heme oxidase-1 (HO-1) and mitogen-activated protein kinases (MAPKs) as well as the corresponding mRNA. Luciferase assays were conducted to evaluate the interaction between NF-E2-related factor-2 (Nrf-2), the antioxidant response element (ARE), and the promoter region of the HO-1 gene and subsequent gene expression. Biochemical markers for hepatotoxicity were monitored to assess whether GDT protected the cells from the t-BHP-mediated oxidative stimuli. RESULTS: GDT induced HO-1 expression via the activation of Nrf-2 nuclear translocation and the subsequent transcription of the HO-1 gene in vitro and in vivo, which seemed to be regulated by phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) and p38 signaling pathways. GDT exhibited in vitro and in vivo hepatoprotective activity as determined by the lowered levels of hepatic enzymes and malondialdehydes and the elevated glutathione levels. CONCLUSIONS: This study validates the ethnopharmacological application of Ganoderma lucidum as a treatment for hepatic disorders. GDT induced in vitro and in vivo anti-inflammatory activity in t-BHP-damaged hepatic cells through the expression of HO-1, and in which PI3K/Akt and p38 kinases are involved. Our study motivates further research in the exploration of potent hepatoprotective agents from Ganoderma lucidum.


Assuntos
Lanosterol/análogos & derivados , Estresse Oxidativo/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Alanina Transaminase/metabolismo , Animais , Aspartato Aminotransferases/metabolismo , Linhagem Celular Tumoral , Doença Hepática Induzida por Substâncias e Drogas/tratamento farmacológico , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Frutas , Ganoderma , Glutationa/metabolismo , Heme Oxigenase-1/genética , Heme Oxigenase-1/metabolismo , Lanosterol/farmacologia , Lanosterol/uso terapêutico , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Endogâmicos ICR , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Extratos Vegetais , Substâncias Protetoras/uso terapêutico , Proteínas Proto-Oncogênicas c-akt/metabolismo , terc-Butil Hidroperóxido
12.
Arch Plast Surg ; 40(1): 52-6, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23362480

RESUMO

BACKGROUND: Measuring grip and pinch strength is an important part of hand injury evaluation. Currently, there are no standardized values of normal grip and pinch strength among the Korean population, and lack of such data prevents objective evaluation of post-surgical recovery in strength. This study was designed to establish the normal values of grip and pinch strength among the healthy Korean population and to identify any dependent variables affecting grip and pinch strength. METHODS: A cross-sectional study was carried out. The inclusion criterion was being a healthy Korean person without a previous history of hand trauma. The grip strength was measured using a Jamar dynamometer. Pulp and key pinch strength were measured with a hydraulic pinch gauge. Intra-individual and inter-individual variations in these variables were analyzed in a standardized statistical manner. RESULTS: There were a total of 336 healthy participants between 13 and 77 years of age. As would be expected in any given population, the mean grip and pinch strength was greater in the right hand than the left. Male participants (137) showed mean strengths greater than female participants (199) when adjusted for age. Among the male participants, anthropometric variables correlated positively with grip strength, but no such correlations were identifiable in female participants in a statistically significant way. CONCLUSIONS: Objective measurements of hand strength are an important component of hand injury evaluation, and population-specific normative data are essential for clinical and research purposes. This study reports updated normative hand strengths of the South Korean population in the 21st century.

13.
J Org Chem ; 77(20): 9156-62, 2012 Oct 19.
Artigo em Inglês | MEDLINE | ID: mdl-23009711

RESUMO

A simple, mild, and inexpensive biphasic functionalization approach is attempted for preparing an ideal core-shell-type resin. The core-shell-type architecture was constructed by coupling Fmoc-OSu to the amino groups on the shell layer of an aminomethyl polystyrene (AM PS) resin. The shell layer thickness of the resin could be easily controlled under mild conditions, which was characterized by confocal laser scanning microscopy (CLSM). The efficiency of core-shell-type resin for solid-phase peptide synthesis (SPPS) was demonstrated by the synthesis of various peptides and compared with commercially available noncore-shell-type resins such as AM PS and poly(ethylene glycol)-based resins. The core-shell-type resin provided effective performance during the synthesis of hydrophobic peptide sequences, a disulfide-bridged cyclic peptide, and a difficult PNA sequence. Furthermore, a highly aggregative peptide fragment, MoPrP 105-125, was synthesized more efficiently on the core-shell-type resin under microwave conditions than AM PS and ChemMatrix resins.


Assuntos
Peptídeos/síntese química , Poliestirenos/química , Resinas Sintéticas/química , Microscopia Confocal , Peptídeos/química , Polietilenoglicóis/química
14.
Clin Orthop Surg ; 4(2): 134-8, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22662299

RESUMO

BACKGROUND: Not much is known regarding avulsion fractures of the calcaneal tuberosity. We propose a modified classification scheme that presents the four types of calcaneal avulsion fracture as described by surgical and magnetic resonance imaging (MRI) findings, and evaluation of their specific features. METHODS: Out of 764 cases of calcaneal fractures, we examined 20 cases (2.6%) that involved the tuberosity of the calcaneus. Each case was classified depending on the avulsed fracture patterns as follows; type I is a 'simple extra-articular avulsion' fracture, type II is the 'beak' fracture, type III is an infrabursal avulsion fracture from the middle third of the posterior tuberosity, and finally in type IV there is the 'beak', but a small triangular fragment is separated from the upper border of the tuberosity. We examined the features of each avulsed type according to several criteria including patient age, gender, anatomical variances of the Achilles tendon, the fibers involved and the mechanism of injury. RESULTS: The type I fracture (8/20 cases) was the most common and likely to occur in elderly women. However, in other types, they were more common in relatively younger male patients. Type I were usually caused due to an accidental trip causing a fall by the patient. However, the dominant cause of type II (5/20 cases) fractures a direct blow or hit directly to the bone. Type III (4/20 cases) and IV (3/20 cases) fractures were likely to occur due to falling. All fibers within the Achilles tendon are involved in both type I and II fractures. However, only the superficial fibers are involved in type III fractures, whereas the deep fibers are involved in type IV fractures. CONCLUSIONS: The avulsion patterns of the calcaneal tuberosity fractures are the result of several factors including the bony density level, the mechanism of injury and the fibers of the Achilles tendon that transmit the force. Accurate diagnosis of type III and IV is dependant on MRI technology to confirm the specific location of the injury and provide proper patient treatment therapeutics.


Assuntos
Calcâneo/lesões , Fraturas Ósseas/classificação , Adolescente , Adulto , Fatores Etários , Idoso , Calcâneo/diagnóstico por imagem , Calcâneo/patologia , Criança , Feminino , Fraturas Ósseas/diagnóstico por imagem , Fraturas Ósseas/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Radiografia , Estatísticas não Paramétricas
15.
J Control Release ; 162(1): 218-24, 2012 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-22732476

RESUMO

The success of radioimmunotherapy for solid tumors remains elusive due to poor biodistribution and insufficient tumor accumulation, in part, due to the unique tumor microenvironment resulting in heterogeneous tumor antibody distribution. Pulsed high intensity focused ultrasound (pulsed-HIFU) has previously been shown to increase the accumulation of (111)In labeled B3 antibody (recognizes Lewis(y) antigen). The objective of this study was to investigate the tumor penetration and therapeutic efficacy of pulsed-HIFU exposures combined with (90)Y labeled B3 mAb in an A431 solid tumor model. The ability of pulsed-HIFU (1 M Hz, spatial averaged temporal peak intensity=2685 W cm(-2); pulse repetition frequency=1 Hz; duty cycle=5%) to improve the tumor penetration and therapeutic efficacy of (90)Y labeled B3 mAb ((90)Y-B3) was evaluated in Le(y)-positive A431 tumors. Antibody penetration from the tumor surface and blood vessel surface was evaluated with fluorescently labeled B3, epi-fluorescent microscopy, and custom image analysis. Tumor size was monitored to determine treatment efficacy, indicated by survival, following various treatments with pulsed-HIFU and/or (90)Y-B3. The pulsed-HIFU exposures did not affect the vascular parameters including microvascular density, vascular size, and vascular architecture; although 1.6-fold more antibody was delivered to the solid tumors when combined with pulsed-HIFU. The distribution and penetration of the antibodies were significantly improved (p-value<0.05) when combined with pulsed-HIFU, only in the tumor periphery. Pretreatment with pulsed-HIFU significantly improved (p-value<0.05) survival over control treatments.


Assuntos
Anticorpos Monoclonais/administração & dosagem , Anticorpos Monoclonais/uso terapêutico , Imunoconjugados/administração & dosagem , Imunoconjugados/uso terapêutico , Neoplasias/terapia , Animais , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/farmacocinética , Linhagem Celular Tumoral , Humanos , Imunoconjugados/imunologia , Imunoconjugados/farmacocinética , Camundongos , Camundongos Nus , Neoplasias/imunologia , Neoplasias/patologia , Radioimunoterapia/métodos , Transplante Heterólogo , Terapia por Ultrassom/métodos , Radioisótopos de Ítrio/administração & dosagem , Radioisótopos de Ítrio/imunologia , Radioisótopos de Ítrio/farmacocinética , Radioisótopos de Ítrio/uso terapêutico
16.
Nucl Med Biol ; 39(4): 472-83, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22172384

RESUMO

INTRODUCTION: This study was undertaken to investigate the effect of paclitaxel and bevacizumab on the therapeutic efficacy of (90)Y-labeled B3 monoclonal antibody, directed against Le(y) antigen, for the treatment of Le(y)-positive A431 tumors implanted subcutaneously in the right hind flank of nude mice. METHODS: When the tumor size reached ~200 mm(3), the mice received a single dose of intravenous (iv) (90)Y-labeled B3 (60 µCi/150 µg or 100 µCi/150 µg B3), intraperitoneal paclitaxel (40 mg/kg) or iv bevacizumab (5 mg/kg) for monotherapy. To investigate the effect of combined therapies on survival, the mice were treated with two or three agents in the following combinations: (90)Y-B3 on day 0 and paclitaxel on day 1; bevacizumab on -1 day and (90)Y-B3 on day 0; bevacizumab on -1 day and paclitaxel on day 1; bevacizumab, (90)Y-B3 and paclitaxel each at 1-day intervals. The mice with no treatment were used as a control. The tumor volume at 1000 mm(3) was used as a surrogate end point of survival. RESULTS: Compared to control animals, paclitaxel delayed tumor growth with a significantly longer median survival time (P<.001), whereas bevacizumab alone showed a less pronounced effect on a median survival time (P=.18). (90)Y-B3 increased the median survival time in a dose-dependent manner (P<.05). The combined therapy of bevacizumab with paclitaxel produced a trend toward an increase of the median survival time compared to paclitaxel alone (P=.06), whereas bevacizumab combined with (90)Y-B3 showed a statistically insignificant increase in the median survival time compared to (90)Y-B3 alone (P=.25). The tumor sizes of all animals in these groups reached the surrogate end point of survival by day 35. In contrast, the combined therapy involving (90)Y-B3 with paclitaxel showed a striking synergistic effect in shrinking tumors and prolonging the survival time (P<.001); on day 120, three of nine mice (33%) and six of six mice (100%) were alive without tumor when treated with 60 µCi (90)Y-B3 and 100 µCi (90)Y-B3, respectively. The addition of bevacizumab treatment 1 day before the combined therapy of 60 µCi (90)Y-B3 with paclitaxel did not produce a statistically significant increase in survival when compared to the (90)Y-B3 with paclitaxel (P>.10). Fluorescence microscopy analysis indicated that paclitaxel increased, whereas bevacizumab decreased, the accumulation and penetration of Alexa Fluor 647-B3 into tumor microenvironment compared to the control (P<.05). CONCLUSION: Our findings on the paclitaxel effect support a hypothesis that the increased tumor accumulation and penetration of (90)Y-B3 as well as the high radiosensitization of tumor cells by paclitaxel may be the major factors responsible for the synergistic effect of the combined therapy involving (90)Y-B3 with paclitaxel.


Assuntos
Anticorpos Monoclonais Humanizados/farmacologia , Paclitaxel/farmacologia , Radioimunoterapia/métodos , Animais , Anticorpos Monoclonais Humanizados/uso terapêutico , Bevacizumab , Carcinoma de Células Escamosas/irrigação sanguínea , Carcinoma de Células Escamosas/tratamento farmacológico , Carcinoma de Células Escamosas/patologia , Carcinoma de Células Escamosas/radioterapia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/efeitos da radiação , Terapia Combinada , Sinergismo Farmacológico , Humanos , Camundongos , Microscopia de Fluorescência , Microvasos/efeitos dos fármacos , Microvasos/metabolismo , Microvasos/efeitos da radiação , Paclitaxel/uso terapêutico , Ensaios Antitumorais Modelo de Xenoenxerto , Radioisótopos de Ítrio/uso terapêutico
17.
Nucl Med Biol ; 38(8): 1119-27, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21741258

RESUMO

INTRODUCTION: Radiolabeling of a monoclonal antibody (mAb) with a metallic radionuclide requires the conjugation of a bifunctional chelator to the mAb. The conjugation, however, can alter the physical and immunological properties of the mAb, consequently affecting its tumor-targeting pharmacokinetics. In this study, we investigated the effect of the amount of 2-(p-isothiocyanatobenzyl)-cyclohexyl-diethylenetriamine-pentaacetic acid (CHX-A″) conjugated to MORAb-009, a mAb directed against mesothelin, and the effect of MORAb dose on the biodistribution of (111)In-labeled MORAb-009. METHODS: We used nude mice bearing the A431/K5 tumor as a mesothelin-positive tumor model and the A431 tumor as a mesothelin-negative control. To find the optimal level of CHX-A″ conjugation, CHX-A″-MORAb-009 conjugates with 2.4, 3.5 and 5.5 CHX-A″ molecules were investigated. To investigate the effect of injected MORAb-009 dose on neutralizing the shed mesothelin in the circulation, biodistribution studies were performed after the intravenous co-injection of (111)In-labeled MORAb-009 (2.4 CHX-A″/MORAb-009) with three different doses: 0.2, 2 and 30 µg of MORAb-009. RESULTS: The tumor uptake in A431/K5 tumor was four times higher than that in A431 tumor, indicating that the tumor uptake in A431/K5 was mesothelin mediated. The conjugate with 5.5 CHX-A″ showed a lower isoelectric point (pI) and lower immunoreactivity (IR) than the 2.4 CHX-A″ conjugate. These differences were reflected in the biodistribution of the (111)In label. The (111)In-labeled MORAb-009 conjugated with 2.4 CHX-A″ produced higher tumor uptake and lower liver and spleen uptakes than the 5.5 CHX-A″ conjugate. The biodistribution studies also revealed that the tumor uptake was significantly affected by the injected MORAb-009 dose and tumor size. The 30-µg dose produced higher tumor uptake than the 0.2- and 2-µg doses, whereas the 30-µg dose produced lower liver and spleen uptakes than the 0.2-µg dose. CONCLUSION: This study demonstrates that the number of chelate conjugation and the injected dose are two important parameters to achieve high tumor and low non-target organ uptake of (111)In-labeled MORAb-009. This study also suggests that the injected dose of mAb could be individualized based on the tumor size or the blood level of shed antigen in a patient to achieve the ideal tumor-to-organ radioactivity ratios.


Assuntos
Anticorpos Monoclonais/farmacocinética , Quelantes/farmacocinética , Proteínas Ligadas por GPI/metabolismo , Radioisótopos de Índio/farmacocinética , Isotiocianatos/farmacocinética , Neoplasias Experimentais/metabolismo , Ácido Pentético/análogos & derivados , Animais , Anticorpos Monoclonais/administração & dosagem , Relação Dose-Resposta à Radiação , Fígado/metabolismo , Mesotelina , Camundongos , Camundongos Nus , Ácido Pentético/farmacocinética , Baço/metabolismo , Distribuição Tecidual
18.
Lab Chip ; 9(18): 2683-90, 2009 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-19704984

RESUMO

We report two types of signal enhancement strategy derived from the origin of mechanical response, surface stress and mass, of the dynamic mode microcantilever for the detection of PSA at low picogram scales (low femtomolar concentration). The PSA detection at extremely low concentration levels is crucial to the early detection of relapses of prostate cancer after the radical prostatectomy and the detection of breast cancer in patient's serum. There is a clear need for the ultrasensitive detection of PSA via simple and rapid diagnostic tools. From the motives, to increase the sensitivity of the microcantilever, PSA polyclonal antibody (PSA pAb) as an additional surface stress inducer and PSA polyclonal antibody-conjugated silica nanoparticles (pAb-SiNPs) as a mass inducer have been applied to the PSA-captured microcantilevers. From two types of sandwich assay, we could confirm the sensitivity enhancement effects (2 approximately 4 times enhanced at the same concentrations) enough to detect PSA at low picogram levels (LOD of 1 pg/mL or below). Moreover, surface stress due to steric interactions between epitope-specific monoclonal antibodies was assessed to support a signal amplification strategy by stress inducer, and the reduction of signal enhancement due to stiffness increase by the mass inducer was studied to clarify the sensitivity enhancement of the microcantilever by mass inducer.


Assuntos
Antígeno Prostático Específico/análise , Anticorpos Monoclonais/química , Campos Eletromagnéticos , Humanos , Masculino , Nanopartículas , Nanotecnologia , Prostatectomia , Dióxido de Silício , Estresse Mecânico , Propriedades de Superfície
19.
Arch Pharm Res ; 32(5): 655-9, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-19471878

RESUMO

The antimicrobial effect of solvent extracts from the rhizome of a thick-stemmed wood fern (Dryopteris crassirhizoma) was evaluated and its phloroglucinol components, flavaspidic acids PB and AB. Flavaspidic acids PB and AB were isolated from the D. crassirhizoma rhizomes by methanol extraction, followed by silica gel and Sephadex LH-20 column chromatography. The chemical structures were characterized by spectral techniques, including ESI-MS, UV, (1)H- and (13)C-NMR spectrum analysis. When the antimicrobial activity of the extracts and compounds was tested by the paper disc method, the extracts and compounds were highly active against Gram-positive bacteria, such as methicillin-resistant Staphylococcus aureus KCTC 1928 (a MRSA bacterium), Streptococcus mutans and Bacillus subtilis. The extracts and compounds were not active against fungi and chlorella. Our study revealed that the antibacterial activity of samples from D. crassirhizoma was mainly related to the flavaspidic acids.


Assuntos
Antibacterianos/farmacologia , Butirofenonas/farmacologia , Dryopteris/química , Bactérias Gram-Positivas/efeitos dos fármacos , Antibacterianos/química , Antibacterianos/isolamento & purificação , Butirofenonas/química , Butirofenonas/isolamento & purificação , Cromatografia Líquida , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Bactérias Gram-Positivas/crescimento & desenvolvimento , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Rizoma , Espectrometria de Massas por Ionização por Electrospray , Espectrofotometria Ultravioleta
20.
Anal Chem ; 81(3): 1008-15, 2009 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-19117480

RESUMO

Immunoassays using nanomaterials have been rapidly developed for the analysis of multiple biomolecules. Highly sensitive and biocompatible surface enhanced Raman spectroscopy-active nanomaterials have been used for biomolecule analysis by many research groups in order to overcome intrinsic problems of conventional immunoassays. We used fluorescent surface-enhanced Raman spectroscopic dots (F-SERS dots) to detect biomolecules in this study. The F-SERS dots are composed of silver nanoparticle-embedded silica nanospheres, organic Raman tagging materials, and fluorescent dyes. The F-SERS dots demonstrated highly sensitive, selective, and multifunctional characteristics for multiplex targeting, tracking, and imaging of cellular and molecular events in the living organism. We successfully applied F-SERS dots for the detection of three cellular proteins, including CD34, Sca-1, and SP-C. These proteins are simultaneously expressed in bronchioalveolar stem cells (BASCs) in the murine lung. We analyzed the relative expression ratios of each protein in BASCs since external standards were used to evaluate SERS intensity in tissue. Quantitative comparisons of multiple protein expression in tissue were first attempted using SERS-encoded nanoprobes. Our results suggested that immunoassays using F-SERS dots offered significant increases in sensitivity and selectivity. Such immunoassays may serve as the primary next-generation labeling technologies for the simultaneous analysis of multiple biomolecules.


Assuntos
Corantes Fluorescentes/química , Imunoensaio/métodos , Nanopartículas/química , Alvéolos Pulmonares/citologia , Análise Espectral Raman/métodos , Células-Tronco/metabolismo , Animais , Anticorpos/química , Antígenos CD34/análise , Antígenos Ly/análise , Linhagem Celular Tumoral , Peptídeos e Proteínas de Sinalização Intercelular , Proteínas de Membrana/análise , Camundongos , Peptídeos/análise , Proteína C Associada a Surfactante Pulmonar
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