RESUMEN
OBJECTIVES: Extracellular matrix components play a significant role in maintaining tissue integrity and pathological processes of the temporomandibular joint (TMJ). This study aimed to evaluate the influence of a soft diet on the mRNA expression of proteoglycans and glycosaminoglycans (GAGs) linked to proteoglycan core proteins in rat TMJ discs. METHODS: Thirty 4-week-old male Wistar rats were assigned to one of two groups: a control group fed a regular pellet diet and a soft diet group fed a powdered diet for 4 weeks. The mRNA expression levels of 12 proteoglycans in TMJ discs were evaluated using real-time polymerase chain reaction (PCR). In addition, histomorphometric and biochemical analyses were performed to evaluate the thickness and deoxyribonucleic acid (DNA), GAG, and water content of the TMJ discs. RESULTS: The TMJ disc thickness in the anterior, intermediate, and posterior bands decreased significantly in the soft diet group. The GAG content decreased significantly in the soft-diet group, whereas no significant differences in DNA content or water content ratio were observed between the groups. Real-time PCR indicated that the expression levels of aggrecan, versican, biglycan, decorin, fibromodulin, lumican, and chondroadherin decreased in the soft diet group. The expression levels of all versican isoforms decreased in the soft diet group. CONCLUSIONS: These results indicate that the biomechanical environment of the TMJ caused by a soft diet is closely related to the expression of proteoglycans in TMJ discs, which may eventually increase the fragility of the TMJ discs.
Asunto(s)
Proteoglicanos , Ratas Wistar , Disco de la Articulación Temporomandibular , Animales , Proteoglicanos/metabolismo , Proteoglicanos/genética , Ratas , Masculino , Disco de la Articulación Temporomandibular/metabolismo , Disco de la Articulación Temporomandibular/patología , Reacción en Cadena en Tiempo Real de la Polimerasa , Dieta/efectos adversos , Glicosaminoglicanos/metabolismo , ARN Mensajero/metabolismo , ARN Mensajero/genéticaRESUMEN
Eye drops, including solutions and suspensions, are essential dosage forms to treat ophthalmic diseases, with poorly water-soluble drugs typically formulated as ophthalmic suspensions. In addition to low bioavailability, suspensions exhibit limited efficacy, safety, and usability due to the presence of drug particles. Improving bioavailability can reduce the drug concentrations and the risk of problems associated with suspended drug particles. However, practical penetration enhancers capable of improving bioavailability remain elusive. Herein, we focused on penetratin (PNT), a cell-penetrating peptide (CPP) that promotes active cellular transport related to macromolecule uptake, such as micropinocytosis. According to the in vitro corneal uptake study using a reconstructed human corneal epithelial tissue model, LabCyte CORNEA-MODEL24, PNT enhanced the uptake of Fluoresbrite® YG carboxylate polystyrene microspheres without covalent binding. In an ex vivo porcine eye model, the addition of 10 µM PNT to rebamipide ophthalmic suspension markedly improved the corneal uptake of rebamipide; however, the addition of 100 µM PNT was ineffective due to potentially increased particle size by aggregation. This article provides basic information on the application of PNT as a penetration enhancer in ophthalmic suspensions, including the in vitro and ex vivo studies mentioned above, as well as the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity assay and storage stability at different pH values.
Asunto(s)
Péptidos de Penetración Celular , Córnea , Soluciones Oftálmicas , Suspensiones , Animales , Péptidos de Penetración Celular/química , Péptidos de Penetración Celular/administración & dosificación , Soluciones Oftálmicas/administración & dosificación , Humanos , Córnea/metabolismo , Córnea/efectos de los fármacos , Porcinos , Quinolonas/administración & dosificación , Quinolonas/farmacocinética , Quinolonas/química , Administración Oftálmica , Disponibilidad Biológica , Epitelio Corneal/efectos de los fármacos , Epitelio Corneal/metabolismo , Tamaño de la Partícula , Alanina/análogos & derivadosRESUMEN
Ocular tissues function as biological barriers that hinder drug delivery, depending on the target tissue and route of administration, and must be overcome to achieve the desired therapeutic effect. Penetration enhancers have long been investigated to improve corneal drug penetration via eye drop instillation; however, further development is warranted owing to potential safety concerns. In the present study, we focused on cell-penetrating peptides (CPPs) as a penetration enhancer to address the requirements and explored CPP candidates suitable for corneal drug delivery. Using a reconstructed human corneal epithelial tissue model, LabCyte CORNEA-MODEL24 as an alternative to animal testing that is expected to have higher reproducibility than extracted eyeballs and octa-arginine (R8) as a representative model CPP with simple structure, we investigated the enhancement of 6-carboxyfluorescein (6-FAM) uptake by fluorescence imaging and the potential of eye irritation by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Also, surface plasmon resonance (SPR) evaluated the interaction between R8 and model compounds, suggesting that the stronger interaction could facilitate the corneal uptake of compounds. A comparative screening study of corneal uptake using various CPPs showed that the CPPs other than R8 also have the potential to enhance the corneal uptake of 6-FAM. In particular, penetratin (PNT) showed stronger fluorescence intensity. Through these findings, this manuscript provides beneficial information for the development of a novel corneal penetration enhancer with CPPs. In the future, it is expected that the basic findings with R8 will be verified to be applicable to other CPPs for development as penetration enhancers for eye drop formulation.
Asunto(s)
Péptidos de Penetración Celular , Animales , Humanos , Péptidos de Penetración Celular/química , Soluciones Oftálmicas , Reproducibilidad de los Resultados , Córnea , Sistemas de Liberación de MedicamentosRESUMEN
When orthodontic forces are applied to teeth, bone remodeling, which consists of bone resorption and bone formation, occurs around the teeth. Transient receptor potential vanilloid 2 (TRPV2) is a cation channel expressed in various cell types that responds to various stimuli, including mechanical stress, and involved in calcium oscillations during the early stages of osteoclast differentiation. However, in vivo expression of TRPV2 in osteoclasts has not yet been reported, and temporo-spatial expression of TRPV2 during osteoclast differentiation is unclear. In this study, we examined the TRPV2 expression during experimental tooth movement and assessed the effect of TRPV2 on osteoclast differentiation. TRPV2 was detected on day 1 after experimental tooth movement on the compression side, and the number of TRPV2-expressing cells significantly increased on day 7. These TRPV2-expressing cells had a single, or multiple nuclei and were positive for TRAP activity. Consistent with these in vivo findings, in vitro experiments using RAW264.7 osteoclast progenitor cells showed that TRPV2 mRNA was increased at the early stage of osteoclast differentiation and maintained until the late stage. Furthermore, a TRPV2 channel selective antagonist significantly inhibited osteoclast differentiation. These findings suggest that TRPV2 may have a regulatory role in osteoclast differentiation during orthodontic tooth movement.
Asunto(s)
Resorción Ósea , Osteoclastos , Animales , Ratas , Remodelación Ósea , Diferenciación Celular , Técnicas de Movimiento DentalRESUMEN
We report an 83-year-old man with myasthenia gravis (MG) who developed respiratory depression after spinal anesthesia for transurethral laser enucleation of the prostate. He became less responsive after complained of dyspnea, with a decrease of SpO2 to 83% approximately 13 min after intrathecal administration of 0.5% isobaric bupivacaine 3 ml. With a diagnosis of exacerbation of MG, hydrocortisone 100 mg was administered, following which both consciousness and spontaneous respiration rapidly improved. Cold sense was observed below the C4 dermatome. We provided general anesthesia without using muscle relaxants until disappearance of the effect of spinal anesthesia. Surgery completed uneventfully and confirmed wearing off the local anesthetics effect. He was discharged without respiratory problems on postoperative 3 day.
Asunto(s)
Anestesia Raquidea , Miastenia Gravis , Anciano de 80 o más Años , Anestesia Raquidea/efectos adversos , Anestésicos Locales , Bupivacaína/efectos adversos , Humanos , Masculino , Miastenia Gravis/complicaciones , EsteroidesRESUMEN
The olfactory systems of insects are fundamental to all aspects of their behaviour, and insect olfactory receptor neurons (ORNs) exhibit exquisite specificity and sensitivity to a wide range of environmental cues. In Drosophila melanogaster, ORN responses are determined by three different receptor families, the odorant (Or), ionotropic-like (IR) and gustatory (Gr) receptors. However, the precise mechanisms of signalling by these different receptor families are not fully understood. Here we report the unexpected finding that the type 4 P-type ATPase phospholipid transporter dATP8B, the homologue of a protein associated with intrahepatic cholestasis and hearing loss in humans, is crucial for Drosophila olfactory responses. Mutations in dATP8B severely attenuate sensitivity of odorant detection specifically in Or-expressing ORNs, but do not affect responses mediated by IR or Gr receptors. Accordingly, we find dATP8B to be expressed in ORNs and localised to the dendritic membrane of the olfactory neurons where signal transduction occurs. Localisation of Or proteins to the dendrites is unaffected in dATP8B mutants, as is dendrite morphology, suggesting instead that dATP8B is critical for Or signalling. As dATP8B is a member of the phospholipid flippase family of ATPases, which function to determine asymmetry in phospholipid composition between the outer and inner leaflets of plasma membranes, our findings suggest a requirement for phospholipid asymmetry in the signalling of a specific family of chemoreceptor proteins.
Asunto(s)
Proteínas de Drosophila/genética , Neuronas Receptoras Olfatorias/metabolismo , Proteínas de Transferencia de Fosfolípidos/genética , Receptores Odorantes/genética , Olfato/genética , Animales , Células Quimiorreceptoras/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster , Neuronas Receptoras Olfatorias/fisiología , Proteínas de Transferencia de Fosfolípidos/metabolismo , Receptores Odorantes/metabolismo , Transducción de SeñalRESUMEN
We previously reported that compound 1, having a similar conformation to PTK787 (2) by forming a pseudo ring structure with an intramolecular non-bonded S-O interaction, exhibited a potent inhibitory activity against VEGFR2 tyrosine kinase (KDR). Applying the ideas of pseudo ring formations, we have designed three types of novel indole carboxamide derivatives 5-7 with an intramolecular hydrogen bonding or non-bonded S-O interaction. We describe the design and synthesis of 5-7, and also discuss the relationships of their KDR inhibitory activity and conformations that were stabilized by their intramolecular non-bonded interactions.
Asunto(s)
Inhibidores Enzimáticos/química , Indoles/química , Receptor 2 de Factores de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Inhibidores Enzimáticos/farmacología , Indoles/farmacología , Modelos Moleculares , Estructura MolecularRESUMEN
Optimization of compounds 5 and 6 led to the discovery of VEGF inhibitor 10g which reduced CYP inhibition. It was highly active in vitro (VEGF induced HUVEC proliferation assay) and showed efficacies in three disease models in vivo (cancer, RA, and AMD).
Asunto(s)
Inhibidores de la Angiogénesis/química , Piridinas/química , Factor A de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Inhibidores de la Angiogénesis/síntesis química , Inhibidores de la Angiogénesis/uso terapéutico , Animales , Artritis Reumatoide/tratamiento farmacológico , Células Cultivadas , Inhibidores Enzimáticos del Citocromo P-450 , Sistema Enzimático del Citocromo P-450/metabolismo , Humanos , Ratones , Neoplasias/tratamiento farmacológico , Neovascularización Patológica/tratamiento farmacológico , Trasplante Heterólogo , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
Optimization from compound 4a, having intramolecular S-O nonbonded interaction, led to discover compounds 4m and 4n. They were highly active in vitro (VEGF induced HUVEC proliferation assay) and showed efficacies in three disease models in vivo (cancer, RA, AMD).
Asunto(s)
Anilidas/química , Piridinas/química , Factor A de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Anilidas/síntesis química , Anilidas/uso terapéutico , Animales , Artritis Experimental/tratamiento farmacológico , Línea Celular , Neovascularización Coroidal/tratamiento farmacológico , Modelos Animales de Enfermedad , Células Endoteliales/citología , Humanos , Ratones , Neoplasias/tratamiento farmacológico , Piridinas/síntesis química , Piridinas/uso terapéutico , Ratas , Relación Estructura-Actividad , Factor A de Crecimiento Endotelial Vascular/metabolismo , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
We designed and synthesized a novel 1,4-benzoxazin-3-one derivative 4 which would have inhibitory activities against tyrosine kinases. They could be synthesized easily from various carboxylic acids 10 and commercially available amines using TFP resin without purification. In this article, we will report the design and synthesis of a novel 1,4-benzoxazin-3-one chemical library 4 and the inhibitory activities against KDR and ABL which are closely related to chronic diseases such as cancer.
Asunto(s)
Benzoxazinas/síntesis química , Proteínas Tirosina Quinasas/antagonistas & inhibidores , Aminas , Benzoxazinas/farmacología , Ácidos Carboxílicos , Diseño de Fármacos , Proteínas Proto-Oncogénicas c-abl/antagonistas & inhibidores , Bibliotecas de Moléculas Pequeñas/síntesis química , Receptor 2 de Factores de Crecimiento Endotelial Vascular/antagonistas & inhibidoresRESUMEN
We found 4-pyridylmethylthio derivative 1 to be very effective in using antiangiogenesis activity to prevent proliferation of HUVECs (Human Umbilical Vein Endothelial Cells), which was induced by vascular endothelial growth factor (VEGF). Compound 1 was equally effective in inhibiting VEGF receptor2 tyrosine kinase (KDR, IC(50)=26nM). We deduced that the inhibition was the result of binding the catalytic domain of VEGF receptor2 tyrosine kinase in a similar fashion to both phthalazine derivative PTK787 2 and anthranylamide derivative AAL993 3. In this report, we will describe the conformational analyses, from ab initio MO calculation and X-ray crystallographic analyses, of compound 1 and the analogs, which include non-active 9, all in comparison with 2 and 3. The conformation-activity relationships suggest that a nonbonded intramolecular interaction between the sulfur and the carbonyl oxygen of 1 was very important in inhibiting KDR.
Asunto(s)
Inhibidores de la Angiogénesis/farmacología , Endotelio Vascular/efectos de los fármacos , Piridinas/farmacología , Venas Umbilicales/efectos de los fármacos , Factor A de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Algoritmos , Inhibidores de la Angiogénesis/síntesis química , Dominio Catalítico , Cristalografía por Rayos X , Relación Dosis-Respuesta a Droga , Endotelio Vascular/citología , Humanos , Concentración 50 Inhibidora , Conformación Molecular , Ftalazinas/farmacología , Piridinas/síntesis química , Relación Estructura-Actividad , Venas Umbilicales/citología , Receptor 2 de Factores de Crecimiento Endotelial Vascular/antagonistas & inhibidoresRESUMEN
EAT/mcl-1 (EAT), a bcl-2 related anti-apoptotic gene, is up-regulated at the early stage of differentiation of human embryonal carcinoma cells; cells which serve as a model for early embryogenesis. We generated transgenic mice for the human EAT gene driven by the EF1 alpha promoter in order to elucidate its functional role in vivo. Histologically, these mice exhibited hyperplasia of Langerhans islet cells; pancreatic cell regions composed of both insulin- and glucagon-producing cells. Furthermore, Bax and Bag-1 -- possible heterodimeric partners for EAT in the anti-apoptotic process -- were up-regulated in islets isolated from the EAT transgenic mice. The insulin tolerance test exhibited no significant difference between the EAT transgenic mice and non-transgenic mice, indicating that islet cell hyperplasia was not due to insulin resistance. In conclusion, EAT transgenic mice exhibit hyperplasia of pancreatic beta cells. EAT may inhibit apoptosis of beta cells, allowing these cells to circumvent the process of apoptosis until the adult stage.
Asunto(s)
Hiperplasia/etiología , Islotes Pancreáticos/patología , Proteínas de Neoplasias/fisiología , Proteínas Proto-Oncogénicas c-bcl-2 , Adenoma de Células de los Islotes Pancreáticos/patología , Animales , Apoptosis , Tamaño de la Célula , Genes bcl-2 , Humanos , Hiperplasia/patología , Insulina/metabolismo , Secreción de Insulina , Islotes Pancreáticos/metabolismo , Hígado/patología , Ratones , Ratones Transgénicos , Proteína 1 de la Secuencia de Leucemia de Células Mieloides , Proteínas de Neoplasias/genéticaRESUMEN
Electrothermal vaporization with two filaments was studied for low-pressure helium-ICP-MS. A 10-microl volume of sample was dried and vaporized, and then introduced to the ICP. With this technique, four elements were simultaneously determined with the acceptable precisions (RSD ca. 10%). The detection limit based on 3sigma of background signals approached the fractional ppb range. The proposed method was successfully applied to the analysis of the certified reference material for human hair.