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1.
Clin Cancer Res ; 30(8): 1595-1606, 2024 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-38593226

RESUMO

PURPOSE: CD137 is a T- and NK-cell costimulatory receptor involved in consolidating immunologic responses. The potent CD137 agonist urelumab has shown clinical promise as a cancer immunotherapeutic but development has been hampered by on-target off-tumor toxicities. A CD137 agonist targeted to the prostate-specific membrane antigen (PSMA), frequently and highly expressed on castration-resistant metastatic prostate cancer (mCRPC) tumor cells, could bring effective immunotherapy to this immunologically challenging to address disease. EXPERIMENTAL DESIGN: We designed and manufactured CB307, a novel half-life extended bispecific costimulatory Humabody VH therapeutic to elicit CD137 agonism exclusively in a PSMA-high tumor microenvironment (TME). The functional activity of CB307 was assessed in cell-based assays and in syngeneic mouse antitumor pharmacology studies. Nonclinical toxicology and toxicokinetic properties of CB307 were assessed in a good laboratory practice (GLP) compliant study in cynomolgus macaques. RESULTS: CB307 provides effective CD137 agonism in a PSMA-dependent manner, with antitumor activity both in vitro and in vivo, and additional activity when combined with checkpoint inhibitors. A validated novel PSMA/CD137 IHC assay demonstrated a higher prevalence of CD137-positive cells in the PSMA-expressing human mCRPC TME with respect to primary lesions. CB307 did not show substantial toxicity in nonhuman primates and exhibited a plasma half-life supporting weekly clinical administration. CONCLUSIONS: CB307 is a first-in-class immunotherapeutic that triggers potent PSMA-dependent T-cell activation, thereby alleviating toxicologic concerns against unrestricted CD137 agonism.


Assuntos
Neoplasias de Próstata Resistentes à Castração , Masculino , Humanos , Camundongos , Animais , Neoplasias de Próstata Resistentes à Castração/tratamento farmacológico , Neoplasias de Próstata Resistentes à Castração/patologia , Imunoterapia/métodos , Microambiente Tumoral
2.
RSC Adv ; 13(15): 9781-9787, 2023 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-36998516

RESUMO

Bisphenol A (BPA) is a common chemical used in plastic production. BPA, which has the potential to be poisonous to plants, has lately emerged as a serious environmental concern owing to its extensive usage and release patterns. Prior study has only looked at how BPA affects plants up to a certain stage in their growth. The precise mechanism of toxicity, penetration of BPA, and damage to internal root tissues remains unknown. Therefore, the goal of this study was to examine the hypothesized mechanism for BPA-induced root cells by studying the effects of bisphenol A (BPA) on the ultrastructure and function of root tip cells of soybean plants. We looked at plant changes in root cell tissues after BPA exposure. Further, the biological characteristics that responded to BPA stress were investigated, and the accumulation of BPA in the root, stem, and leaf of the soybean plant was systematically investigated by using FTIR and SEM analysis. The uptake of BPA is a key internal factor that contributes to changes in biological characteristics. Our findings provide insight into how BPA could alter plant root growth, which might contribute new knowledge toward a better scientific appraisal of the possible dangers of BPA exposure for plants.

3.
Environ Pollut ; 267: 115498, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32905901

RESUMO

Industrial dye effluents, which are a major wastage component that enter the natural environment, pose a significant health risk to human and aquatic life. Therefore, the effective removal of dye effluents is a major concern. Against this backdrop, in this study, a low-cost, earth-abundant, and ecofriendly ɤ-Fe2O3-PPy nanocomposite was prepared employing the conventional hydrothermal method. The morphology, functional groups, and elemental composition of ɤ-Fe2O3-PPy were characterized by XRD, SEM, XPS, and FTIR studies. Under optimized conditions, the prepared novel ɤ-Fe2O3-PPy nanocomposite showed a high methylene blue (MB) adsorption capacity of 464 mg/g, which is significantly higher than that of existing adsorbents such as CNTs and polymer-modified CNTs. The adsorption parameters such as pH, adsorbent dosage, and ionic strength were optimized to enhance the MB adsorption capacity. The adsorption results revealed that MB is adsorbed onto the adsorbent surface via electrostatic interactions, hydrogen bonding, and chemical binding interactions. In terms of practical application, the adsorbent's adsorption-desorption ability in conjunction with magnetic separation was investigated; the prepared ɤ-Fe2O3-PPy nanocomposite exhibited excellent adsorption and desorption efficiencies over more than seven adsorption-desorption cycles.


Assuntos
Nanocompostos , Poluentes Químicos da Água , Purificação da Água , Adsorção , Humanos , Cinética , Águas Residuárias , Poluentes Químicos da Água/análise
4.
J Alzheimers Dis ; 77(1): 1-14, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32741816

RESUMO

Alzheimer's disease (AD) is the most common progressive neurodegenerative disease worldwide, but its cause remains unclear. Although a few drugs can provide temporary and partial relief of symptoms in some patients, no curative treatment is available. Therefore, attention has been focused on research using stem cells to treat AD. Among stem cells, mesenchymal stem cells (MSCs) have been used to treat the related pathologies in animal models of AD, and other neurodegenerative disease. This review describes latest research trends on the use of MSC-based therapies in AD and its action of mechanism. MSCs have several beneficial effects. They would be specified as the reduction of neuroinflammation, the elimination of amyloid-ß, neurofibrillary tangles, and abnormal protein degradation, the promotion of autophagy-associated and blood-brain barrier recoveries, the upregulation of acetylcholine levels, improved cognition, and the recovery of mitochondrial transport. Therefore, this review describes the latest research trends in MSC-based therapy for AD by demonstrating the importance of MSC-based therapy and understanding of its mechanisms in AD and discusses the limitations and perspectives of stem cell therapy in AD.


Assuntos
Doença de Alzheimer/metabolismo , Doença de Alzheimer/terapia , Transplante de Células-Tronco Mesenquimais/tendências , Células-Tronco Mesenquimais/metabolismo , Doença de Alzheimer/genética , Peptídeos beta-Amiloides/genética , Peptídeos beta-Amiloides/metabolismo , Precursor de Proteína beta-Amiloide/genética , Precursor de Proteína beta-Amiloide/metabolismo , Animais , Previsões , Humanos , Transplante de Células-Tronco Mesenquimais/métodos , Emaranhados Neurofibrilares/genética , Emaranhados Neurofibrilares/metabolismo
5.
Molecules ; 25(12)2020 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-32580297

RESUMO

Kirsten rat sarcoma viral oncogene homolog (KRAS)-driven colorectal cancer (CRC) is notorious to target with drugs and has shown ineffective treatment response. The seeds of Pharbitis nil, also known as morning glory, have been used as traditional medicine in East Asia. We focused on whether Pharbitis nil seeds have a suppressive effect on mutated KRAS-driven CRC as well as reserving muscle cell functions during CRC progression. Seeds of Pharbitis nil (Pharbitis semen) were separated by chromatography and the active compound of Pharbitis semen (PN) was purified by HPLC. The compound PN efficiently suppressed the proliferation of mutated KRAS-driven CRC cells and their clonogenic potentials in a concentration-dependent manner. It also induced apoptosis of SW480 human colon cancer cells and cell cycle arrest at the G2/M phase. The CRC related pathways, including RAS/ERK and AKT/mTOR, were assessed and PN reduced the phosphorylation of AKT and mTOR. Furthermore, PN preserved muscle cell proliferation and myotube formation in cancer conditioned media. In summary, PN significantly suppressed mutated KRAS-driven cell growth and reserved muscle cell function. Based on the current study, PN could be considered as a promising starting point for the development of a nature-derived drug against KRAS-mutated CRC progression.


Assuntos
Proliferação de Células/efeitos dos fármacos , Neoplasias Colorretais/tratamento farmacológico , Ipomoea nil/química , Proteínas Proto-Oncogênicas p21(ras)/genética , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Cromatografia Líquida de Alta Pressão , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Humanos , Células Musculares/efeitos dos fármacos , Células Musculares/patologia , Mutação/efeitos dos fármacos , Sementes/química
6.
Foods ; 9(3)2020 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-32106409

RESUMO

An analytical method to measure solubilized orthophosphate ions (HPO42- and PO43- ) from the water-insoluble food additives calcium phosphate dibasic (DCP) and calcium phosphate tribasic (TCP) in processed foods was optimized by comparing ion chromatography (IC) coupled with DS6 conductivity detector (Cond.) and high-performance liquid chromatography (HPLC) with Evaporative light scattering detector (ELSD) methods. The ion-pairing HPLC method could analyze calcium and phosphate ions successively. However, this method exhibited low reproducibility after approximately 48 hours of measurements. The IC method was established as an effective method of measuring orthophosphate ions with high reproducibility using distilled water and KOH solution as the mobile phase with a Dionex column. Matrix-based limit of detections (LOD) and limit of quantifications (LOQ) for snacks and cereals were estimated in the range of 0.01-0.91 µg/mL and 0.21-2.74 µg/mL, respectively. In inter-day and intra-day tests, the calculated precision (%RSD) and accuracy (recovery %) ranged from 0.5% to 6.6% and 82% to 117%, respectively, in both food samples. The levels of DCP or TCP could be analyzed in various positive food samples, and the developed IC method demonstrated good applicability in the analysis of DCP and TCP in collected processed foods.

7.
Nutrients ; 11(1)2018 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-30577618

RESUMO

Cordyceps militaris is a commonly used medicinal mushroom containing various therapeutic effects such as anti-inflammatory, anti-allergic, and anti-cancer activities. This study examined whether Cordyceps militaris on germinated soybeans (GSC) has a suppressive effect on a v-ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS)-driven colorectal cancer which is notorious for its un-druggable features and the ineffectiveness of conventional therapies against it. GSC extract was prepared and its proximate composition and amino acids were analyzed. The suppressive effects were investigated with the KRAS-driven colorectal cancer cell-line, SW480. SW480 proliferation, clonogenic potential, apoptosis, and the RAS/extracellular signal-regulated kinase (ERK) pathway under the GSC treatment were analyzed by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay, flow cytometry, and Western blot, respectively. An in vivo experiment with the SW480 xenograft mouse model was performed. As a result, GSC suppressed cell proliferation by inducing the apoptosis of KRAS-driven colorectal cancer cells and inhibited clonogenic capabilities. The decrease of KRAS and ERK phosphorylation was detected by Western blot. Tumor growth was significantly suppressed when GSC was introduced to the tumor-xenograft mouse model. In conclusion, GSC suppressed KRAS-driven colorectal cancer growth both in vitro and in vivo, and can be used as an alternative or simultaneous approach in colorectal cancer therapy.


Assuntos
Neoplasias Colorretais/tratamento farmacológico , Cordyceps/química , Inibidores Enzimáticos/farmacologia , Glycine max , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Proteínas Proto-Oncogênicas p21(ras)/fisiologia , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Neoplasias Colorretais/patologia , Neoplasias Colorretais/prevenção & controle , Cordyceps/crescimento & desenvolvimento , MAP Quinases Reguladas por Sinal Extracelular/antagonistas & inibidores , Germinação , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Proteínas Proto-Oncogênicas p21(ras)/antagonistas & inibidores , Ensaios Antitumorais Modelo de Xenoenxerto
8.
Int J Mol Sci ; 19(11)2018 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-30405049

RESUMO

Cordyceps militaris is a medicinal mushroom used to treat immune-related diseases in East Asia. We investigated the anti-inflammatory effect of the extract of C. militaris grown on germinated Rhynchosia nulubilis (GRC) fermented with Pediococcus pentosaceus ON89A isolated from onion (GRC-ON89A) in vivo as well as in vitro. The anti-inflammatory effect of GRC-ON89A was investigated in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The total polyphenol content (TPC) and total flavonoid content (TFC) in the GRC-ON89A ethanol extract were significantly increased compared to that in GRC. GRC-ON89A hexane fraction (GRC-ON89A-Hex) inhibited the release of nitric oxide (NO) compared to that of the LPS-treated control without cytotoxicity in LPS-stimulated RAW 264.7 macrophages. GRC-ON89A-Hex decreased the inducible NO synthase (iNOS), cyclooxygenase 2 (COX2), and tumor necrosis factor (TNF)-α mRNA expression in LPS-stimulated RAW 264.7 macrophages. In addition, pre-treatment with GRC-ON89A-Hex significantly inhibited LPS-stimulated phosphorylation of mitogen-activated protein kinases (MAPKs) and nuclear factor (NF)-κB. To induce allergic contact dermatitis (ACD), 1-fluoro-2, 4-dinitrofluorobenzene (DNFB) was applied to the surface of the right ears of C57BL/6N mice. GRC-ON89A reduced the ear swelling and thickness in DNFB-induced ACD mice. This study demonstrates the potential usefulness of GRC-ON89A as an anti-inflammatory dietary supplement or drug.


Assuntos
Anti-Inflamatórios/uso terapêutico , Cordyceps/química , Dermatite de Contato/tratamento farmacológico , Fermentação , Inflamação/tratamento farmacológico , Pediococcus pentosaceus/metabolismo , Adenosina/análise , Animais , Anti-Inflamatórios/farmacologia , Desoxiadenosinas/análise , Dermatite de Contato/complicações , Dermatite de Contato/patologia , Regulação para Baixo , Flavonoides/análise , Proteínas I-kappa B/metabolismo , Inflamação/complicações , Inflamação/patologia , Mediadores da Inflamação/metabolismo , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Lipopolissacarídeos , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismo , Fosforilação/efeitos dos fármacos , Polifenóis/análise , Células RAW 264.7 , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fator de Transcrição RelA/metabolismo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo
9.
Prog Mol Biol Transl Sci ; 159: 157-176, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30340786

RESUMO

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated system (Cas) is comprised of repetitive bases followed by short fragments of DNA from a previously invading organism that provide immunity to the most prokaryotic organisms. An RNA-dependent spacer is required for CRISPR/Cas9 to recognize the target DNA. Delivery of the CRISPR/Cas9-guide RNA (gRNA) complex to any cell results in modification of the target sequence. The CRISPR/Cas9-mediated genome editing technique is currently in the spotlight and has several research interests, including molecular medicine and agriculture. There are several factors that hinder the delivery of this complex, such as the large size of the plasmid or high dosage of the chemical agent. There are several methods available to deliver CRISPR/Cas9 and its components to the target cells. It includes viral, non-viral and physical methods to deliver plasmid or ribonucleoprotein (RNP) of CRISPR components. But in vivo CRISPR/Cas9 delivery remains challenging to the researchers due to insertional mutagenesis, targeted delivery, immunogenicity, and off-targets. However, studies suggesting that the CRISPR/Cas9-RNP delivery can overcome these hurdles. Here, we review the various methods for delivery of CRISPR/Cas9 and gRNA to several cell lines, highlighting the limitations of each approach, and suggest possible alternative methods.


Assuntos
Sistemas CRISPR-Cas/genética , Técnicas de Transferência de Genes , Edição de Genes , Vetores Genéticos/metabolismo , Humanos , Células-Tronco/metabolismo
10.
Mol Biotechnol ; 60(4): 329-338, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29516417

RESUMO

Programmable nucleases including zinc finger nucleases, transcription activator-like effector nucleases, and clustered regularly interspaced short palindrome repeats (CRISPR)/CRISPR-associated protein have tremendous potential biological and therapeutic applications as novel genome editing tools. These nucleases enable precise modification of the gene of interest by disruption, insertion, or correction. The application of genome editing technology to pluripotent stem cells or hematopoietic stem cells has the potential to remarkably advance the contribution of this technology to life sciences. Specifically, disease models can be generated and effective therapeutics can be developed with great efficiency and speed. Here we review the characteristics and mechanisms of each programmable nuclease. In addition, we review the applications of these nucleases to stem cells for disease therapies and summarize key studies of interest.


Assuntos
Edição de Genes/métodos , Transplante de Células-Tronco/métodos , Células-Tronco/citologia , Sistemas CRISPR-Cas , Endonucleases/metabolismo , Engenharia Genética , Humanos , Nucleases dos Efetores Semelhantes a Ativadores de Transcrição/metabolismo
11.
J Nanosci Nanotechnol ; 18(2): 1232-1236, 2018 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-29448563

RESUMO

In this work, we present the fabrication and characterization of a 2-chloroethyl ethyl sulfide (2-CEES) gas sensor based on ZnO nanoparticles (NPs) synthesized by a hydrothermal method. We confirmed that synthesized ZnO NPs adopt a polycrystalline phase. Partially aggregated ZnO-NPs revealed spherical or ellipsoidal nanocrystalline particles in a size range of 30-50 nm, as observed by field-emission scanning electron microscopy (FE-SEM). The maximum response of the ZnO NPs was 15 at 1 ppm 2-CEES concentration, and a low detection limit of 0.4 ppm was observed at an optimal operating temperature of 250 °C. The lowest response time was 6 s in 20 ppm at 250 °C. The linearity response with correlation coefficient (R2) was 0.9887 at 2-CEE concentrations of 0.4-1 ppm at the operating temperature of 250 °C. The enhanced sensing performance and a decrease in the operating temperature were attributed to a high specific surface area and more active sites in the ZnO NPs after exposure to 2-CEES.

12.
Genome Res ; 2018 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-29326299

RESUMO

KRAS is the most frequently mutated oncogene in human tumors, and its activating mutations represent important therapeutic targets. The combination of Cas9 and guide RNA from the CRISPR-Cas system recognizes a specific DNA sequence and makes a double-strand break, which enables editing of the relevant genes. Here, we harnessed CRISPR to specifically target mutant KRAS alleles in cancer cells. We screened guide RNAs using a reporter system and validated them in cancer cells after lentiviral delivery of Cas9 and guide RNA. The survival, proliferation, and tumorigenicity of cancer cells in vitro and the growth of tumors in vivo were determined after delivery of Cas9 and guide RNA. We identified guide RNAs that efficiently target mutant KRAS without significant alterations of the wild-type allele. Doxycycline-inducible expression of this guide RNA in KRAS-mutant cancer cells transduced with a lentiviral vector encoding Cas9 disrupted the mutant KRAS gene, leading to inhibition of cancer cell proliferation both in vitro and in vivo. Intra-tumoral injection of lentivirus and adeno-associated virus expressing Cas9 and sgRNA suppressed tumor growth in vivo, albeit incompletely, in immunodeficient mice. Expression of Cas9 and the guide RNA in cells containing wild-type KRAS did not alter cell survival or proliferation either in vitro and in vivo. Our study provides a proof-of-concept that CRISPR can be utilized to target driver mutations of cancers in vitro and in vivo.

13.
ACS Biomater Sci Eng ; 4(12): 4163-4173, 2018 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-33418815

RESUMO

Orally administered antisense therapy has been introduced as an effective approach for treating cancer in the gastrointestinal tract. However, its practical application has been limited by the instability of oligonucleotides and their inefficient delivery. To overcome these problems, we synthesized size-dependent, oligonucleotide nanoparticle-patterned chitosan/phytic acid (ODN/CS/PA) capsules with protective shields via a three-step process of self-assembly, nanoparticle encapsulation, and shell formation. The multicompartmental capsule size and oligonucleotide nanoparticle-loading pattern were controlled by applying different potentials during the electrostatic extrusion process used for nanoparticle encapsulation. Over 95% of encapsulated oligonucleotides were protected from nuclease digestion (DNase I) and, depending on their size, showed 40-75% protection against simulated gastric fluid. Their controlled release from capsules correlated with the cellular delivery of released nanoparticles and the inhibition of protein expression in cancer cells. Specifically, large capsules showed approximately 32-fold greater delivery to cancer cells than nonencapsulated nanoparticles. We also confirmed delivery of oligonucleotide nanoparticles to the small intestine and colon of rats following oral administration. These findings demonstrate that the multicompartmental ODN/CS/PA capsules can facilitate efficient oral delivery of oligonucleotides for cancer treatment.

14.
Invest Ophthalmol Vis Sci ; 58(12): 5329-5335, 2017 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-29049735

RESUMO

Purpose: To characterize the clinical and biological properties of biodegradable collagen matrices (BCMs) for possible glaucoma drainage device implantation. Methods: A total of 68 refractory glaucoma eyes, followed up postoperatively for at least 6 months, were consecutively enrolled after retrospective chart review. The BCM-augmented Ahmed valve implantations (BAAVI) using our Ologen-6 and Ologen-7 valves were performed and compared with a conventional method. Complete surgical success was defined as an IOP of ≤21 mm Hg (IOP 1) or ≤17 mm Hg (IOP 2) without antiglaucoma medications. Qualified success was defined as an IOP ≤21 mm Hg with or without antiglaucoma medications. The biological properties of each BCM were assessed by enzymatic degradation rates via collagenase under ocular physiological conditions. Results: The mean ages and preoperative IOPs were similar for the groups. In the conventional, BAAVI with Ologen-6, and BAAVI with Ologen-7 groups, complete success rates with target IOP 1 were 29.2%, 40.0%, and 66.7%; those with target IOP 2 were 12.5%, 30.0%, and 45.8%; qualified success rates were 45.8%, 55.0%, and 75.0%, respectively. The enzymatic degradation rate of Ologen-7 was significantly slower than that of Ologen-6 (12.5 × 10-3 vs. 28.8 × 10-3). Conclusions: The surgical success rate was highest in the Ologen-7 BAAVI group, with the lowest dependency on postoperative antiglaucoma medication use compared with the conventional and Ologen-6 BAAVI groups. The clinical results correlated with the different biological and physicochemical properties based on the degree of enzymatic degradation and on the structural morphology.


Assuntos
Implantes Absorvíveis , Colágeno , Implantes para Drenagem de Glaucoma , Glaucoma/cirurgia , Glicosaminoglicanos , Feminino , Seguimentos , Glaucoma/fisiopatologia , Humanos , Pressão Intraocular/fisiologia , Masculino , Pessoa de Meia-Idade , Implantação de Prótese , Estudos Retrospectivos , Técnicas de Sutura , Tonometria Ocular , Resultado do Tratamento , Acuidade Visual/fisiologia , Cicatrização
15.
Int J Mol Sci ; 18(8)2017 Aug 11.
Artigo em Inglês | MEDLINE | ID: mdl-28800074

RESUMO

Colon cancer is one of the most common types of cancer, and it has recently become a leading cause of death worldwide. Among colon cancers, the v-ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS)-mutated form is notorious for its non-druggable features. Cetuximab, a monoclonal antibody that binds to the epidermal growth factor receptor, has been introduced as an antitumor therapy; however, secondary resistance and side effects significantly limit its effective use in these cancers. In this study, we prepared Phellinuslinteus on germinated brown rice (PBR) extracts to increase the sensitivity of KRAS-mutated colon cancers to cetuximab. The combined treatment of PBR extract and cetuximab suppressed SW480 cell viability/proliferation, with the cells exhibiting altered cellular morphology and clonogenic potential. AnnexinV-fluorescein isothiocyanate/propidium iodide-stained flow cytometry and Western blotting were performed, and PBR extract combined with cetuximab treatment increased apoptosis of the SW480 cells and suppressed their KRAS protein expression. The potential of PBR as a synergistic anticancer agent was further investigated in a tumor-xenografted mouse model. Tumor growth was significantly suppressed with PBR extract and cetuximab co-treatment. In conclusion, PBR increased the sensitivity of KRAS-mutated colon cancer cells to cetuximab, which indicates the potential use of PBR as a medical food against colon cancer.


Assuntos
Antineoplásicos/uso terapêutico , Basidiomycota/química , Produtos Biológicos/uso terapêutico , Extratos Celulares/uso terapêutico , Cetuximab/uso terapêutico , Neoplasias do Colo/tratamento farmacológico , Animais , Antineoplásicos/administração & dosagem , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Basidiomycota/patogenicidade , Produtos Biológicos/administração & dosagem , Produtos Biológicos/farmacologia , Extratos Celulares/administração & dosagem , Extratos Celulares/farmacologia , Proliferação de Células/efeitos dos fármacos , Cetuximab/administração & dosagem , Cetuximab/farmacologia , Neoplasias do Colo/genética , Resistencia a Medicamentos Antineoplásicos , Sinergismo Farmacológico , Células HT29 , Humanos , Masculino , Camundongos , Camundongos Nus , Oryza/microbiologia , Proteínas Proto-Oncogênicas p21(ras)/genética
16.
Front Microbiol ; 8: 799, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28539917

RESUMO

Salting is one of the oldest food preservation techniques. However, salt is also the source of living halophilic microorganisms that may affect human health. In order to determine the microbial communities of commercial salts, an investigation were done using amplicon sequencing approach in four commercial salts: Ethiopian Afdera salt (EAS), Ethiopian rock salt (ERS), Korean Jangpan salt (KJS), and Korean Topan salt (KTS). Using domain-specific primers, a region of the 16S rRNA gene was amplified and sequenced using a Roche 454 instrument. The results indicated that these microbial communities contained 48.22-61.4% Bacteria, 37.72-51.26% Archaea, 0.51-0.86% Eukarya, and 0.005-0.009% unclassified reads. Among bacteria, the communities in these salts were dominated by the phyla Proteobacteria, Bacteroidetes, Actinobacteria, and Firmicutes. Of the archaea, 91.58% belonged to the class Halobacteria, whereas the remaining 7.58, 0.83, and 0.01% were Nanoarchaea, Methanobacteria, and Thermococci, respectively. This comparison of microbial diversity in salts from two countries showed the presence of many archaeal and bacterial genera that occurred in salt samples from one country but not the other. The bacterial genera Enterobacter and Halovibrio were found only in Korean and Ethiopian salts, respectively. This study indicated the occurrence and diversity of halophilic bacteria and archaea in commercial salts that could be important in the gastrointestinal tract after ingestion.

17.
Prev Nutr Food Sci ; 22(1): 50-55, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28401088

RESUMO

Raphanus sativus L. (RS) is a cruciferous vegetable that is widely consumed in Korea. The anticancer activity of leaves of RS (RSL) extract has been investigated; however, no studies focused on its anti-inflammatory effects. Therefore, the aim of the current study was to evaluate the anti-inflammatory effects of RSL extract. In brief, RSL powder was fractionated into n-hexane, chloroform, ethyl acetate, n-butanol, and water-soluble fractions. Lipopolysaccharide (LPS)-stimulated RAW264.7 cells were treated with each fraction for initial screening. It was found that the chloroform fraction significantly inhibited nitric oxide release in LPS-stimulated RAW264.7 cells with a half maximal inhibitory concentration value of 196 µg/mL. In addition, the mRNA and protein expression levels of inducible nitric oxide synthase, measured using reverse transcriptase-polymerase chain reaction and western blotting, respectively, were reduced in a concentration-dependent manner. Moreover, the inflammatory cyclooxygenase-2 enzyme expression decreased. Furthermore, the expression of nuclear factor-kappa B (NF-κB), the key regulator of the transcriptional activation of the inflammatory cytokine genes, was reduced by the RSL chloroform fraction. Therefore, the results of our study suggest that RSL exhibits anti-inflammatory effects in LPS-stimulated macrophages via NF-κB inactivation.

18.
Cell Mol Life Sci ; 74(18): 3335-3346, 2017 09.
Artigo em Inglês | MEDLINE | ID: mdl-28405721

RESUMO

Human pluripotent stem cells comprise induced pluripotent and embryonic stem cells, which have tremendous potential for biological and therapeutic applications. The development of efficient technologies for the targeted genome alteration of stem cells in disease models is a prerequisite for utilizing stem cells to their full potential. Genome editing of stem cells is possible with the help of synthetic nucleases that facilitate site-specific modification of a gene of interest. Recent advances in genome editing techniques have improved the efficiency and speed of the development of stem cells for human disease models. Zinc finger nucleases, transcription activator-like effector nucleases, and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated system are powerful tools for editing DNA at specific loci. Here, we discuss recent technological advances in genome editing with site-specific nucleases in human stem cells.


Assuntos
Edição de Genes , Células-Tronco/metabolismo , Proteína da Polipose Adenomatosa do Colo/genética , Proteína da Polipose Adenomatosa do Colo/metabolismo , Sistemas CRISPR-Cas/genética , Endodesoxirribonucleases/metabolismo , Genoma Humano , Humanos , Receptores CCR5/genética , Receptores CCR5/metabolismo , Células-Tronco/citologia , Nucleases dos Efetores Semelhantes a Ativadores de Transcrição/genética , Nucleases dos Efetores Semelhantes a Ativadores de Transcrição/metabolismo , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Proteínas ras/genética , Proteínas ras/metabolismo
19.
Biotechnol Prog ; 33(4): 1035-1045, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28440027

RESUMO

The CRISPR/Cas9 gene editing system was originally derived from the prokaryotic adaptive immune system mediated by Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated proteins (Cas). The system has been successfully applied to genome editing in eukaryotes and has contributed to remarkable advances in the life sciences, in areas ranging from agriculture to genetic disease therapies. For efficient editing and extending the influence of this system, proper delivery of its components is crucial. Both viral and nonviral delivery methods are reviewed here, along with the advantages and disadvantages of each. In addition, we review ex vivo and in vivo CRISPR/Cas9 applications for disease therapies. Related remarkable studies are highlighted and relevant startup companies and their drug development pipelines are described. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1035-1045, 2017.


Assuntos
Proteínas Associadas a CRISPR/genética , Sistemas CRISPR-Cas/genética , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas/genética , Técnicas de Transferência de Genes , Terapia Genética , Animais , Humanos
20.
Sensors (Basel) ; 17(1)2016 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-28042843

RESUMO

The effect of cleaning the surface of single-walled carbon nanotube (SWNT) networks by thermal and the O2 plasma treatments is presented in terms of NH3 gas sensing characteristics. The goal of this work is to determine the relationship between the physicochemical properties of the cleaned surface (including the chemical composition, crystal structure, hydrophilicity, and impurity content) and the sensitivity of the SWNT network films to NH3 gas. The SWNT networks are spray-deposited on pre-patterned Pt electrodes, and are further functionalized by heating on a programmable hot plate or by O2 plasma treatment in a laboratory-prepared plasma chamber. Cyclic voltammetry was employed to semi-quantitatively evaluate each surface state of various plasma-treated SWNT-based electrodes. The results show that O2 plasma treatment can more effectively modify the SWNT network surface than thermal cleaning, and can provide a better conductive network surface due to the larger number of carbonyl/carboxyl groups, enabling a faster electron transfer rate, even though both the thermal cleaning and the O2 plasma cleaning methods can eliminate the organic solvent residues from the network surface. The NH3 sensors based on the O2 plasma-treated SWNT network exhibit higher sensitivity, shorter response time, and better recovery of the initial resistance than those prepared employing the thermally-cleaned SWNT networks.

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