RESUMO
BACKGROUND: Oxidative stress contributes to retina ganglion cells (RGCs) loss in variety of ocular diseases, including ocular trauma, ocular vein occlusion, and glaucoma. Scavenging the excessed reactive oxygen species (ROS) in retinal neurovascular unit could be beneficial to RGCs survival. In this study, a polydopamine (PDA)-based nanoplatform is developed to protect RGCs. RESULTS: The PDA nanoparticles efficiently eliminate multi-types of ROS, protect endothelia and neuronal cells from oxidative damage, and inhibit microglia activation in retinas. In an optic nerve crush (ONC) model, single intravitreal injection of PDA nanoparticles could significantly attenuate RGCs loss via eliminating ROS in retinas, reducing the inflammatory response and maintaining barrier function of retinal vascular endothelia. Comparative transcriptome analysis of the retina implied that PDA nanoparticles improve RGCs survival probably by altering the expression of genes involved in inflammation and ROS production. Importantly, as a versatile drug carrier, PDA nanoparticles could deliver brimonidine (a neuroprotection drug) to synergistically attenuate RGCs loss and promote axon regeneration, thus restore visual function. CONCLUSIONS: The PDA nanoparticle-based therapeutic nanoplatform displayed excellent performance in ROS elimination, providing a promising probability for treating retinal degeneration diseases.
Assuntos
Indóis/uso terapêutico , Nanopartículas/química , Traumatismos do Nervo Óptico/patologia , Polímeros/uso terapêutico , Degeneração Retiniana/tratamento farmacológico , Animais , Tartarato de Brimonidina/química , Tartarato de Brimonidina/farmacologia , Tartarato de Brimonidina/uso terapêutico , Sobrevivência Celular/efeitos dos fármacos , Modelos Animais de Doenças , Células Endoteliais da Veia Umbilical Humana , Humanos , Peróxido de Hidrogênio/farmacologia , Indóis/química , Indóis/farmacologia , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismo , Polímeros/química , Polímeros/farmacologia , Espécies Reativas de Oxigênio/química , Retina/efeitos dos fármacos , Retina/fisiologia , Degeneração Retiniana/patologia , Células Ganglionares da Retina/citologia , Células Ganglionares da Retina/metabolismo , Transcriptoma/efeitos dos fármacosRESUMO
A general strategy to tune the achiral main chain polymers into helical nanoassemblies was proposed based on the co-gelation approach. As an example, two achiral main chain polymers, PCz8 and PSi8, were selected, and their co-assembly with an amphiphilic l- or d-glutamide gelator was investigated. Although the polymers could not form gels individually, they could form co-gels with the gelator and the resultant gels exhibited macroscopic supramolecular chirality, which could be confirmed by CD spectra and SEM observations. Moreover, the supramolecular chirality can be memorized even after the gelator molecules were removed. Remarkably, either the gelator-containing or gelator-free chiral polymer assemblies showed circularly polarized luminescence (CPL), which is usually inherent to intrinsic chiral polymers. It was suggested that during the co-gelation, the chirality of the gelator was transferred to and memorized by the achiral polymers. The approach seems to be general and we provided the first example to tune the achiral polymers into helical assemblies through the co-gelation.
Assuntos
Géis/química , Nanoestruturas/química , Polímeros/química , Glutamina/análogos & derivados , IsomerismoRESUMO
Our aim is to investigate the cytobiological effects of autologous platelet-rich fibrin (PRF) on dental pulp stem cells (DPSCs) and to explore the ectopic and orthotopic possibilities of dental pulp revascularization and pulp-dentin complex regeneration along the root canal cavities of the tooth by using a novel tissue-engineered transplant composed of cell-sheet fragments of DPSCs and PRF granules. Canine DPSCs were isolated and characterized by assaying their colony-forming ability and by determining their cell surface markers and osteogenic/adipogenic differentiation potential. The biological effects of autologous PRF on DPSCs, including cell proliferation, alkaline phosphatase (ALP) activity and odonto-/osteogenic gene expression, were then investigated and quantified. A novel transplant consisting of cell-sheet fragments of DPSCs and PRF granules was adopted to regenerate pulp-dentin-like tissues in the root canal, both subcutaneously in nude mice and in the roots of canines. PRF promoted the proliferation of DPSCs in a dose- and time-dependent manner and induced the differentiation of DPSCs to odonto-/osteoblastic fates by increasing the expression of the Alp, Dspp, Dmp1 and Bsp genes. Transplantation of the DPSC/PRF construct led both to a favorable regeneration of homogeneous and compact pulp-like tissues with abundantly distributed blood capillaries and to the deposition of regenerated dentin along the intracanal walls at 8 weeks post-operation. Thus, the application of DPSC/PRF tissue constructs might serve as a potential therapy in regenerative endodontics for pulp revitalization or revascularization.
Assuntos
Plaquetas/metabolismo , Polpa Dentária/irrigação sanguínea , Polpa Dentária/citologia , Fibrina/metabolismo , Transplante de Células-Tronco , Células-Tronco/citologia , Engenharia Tecidual/métodos , Animais , Materiais Biocompatíveis/metabolismo , Células Cultivadas , Cães , Humanos , Masculino , Camundongos Nus , Odontogênese , Osteogênese , Regeneração , Células-Tronco/metabolismoRESUMO
OBJECTIVE: To explore correlations of 3 novel radiographic parameters with myelopathy induced by posterior atlantoaxial dislocation (PAAD) secondary to os odontoideum (OO) and assess their reproducibility. METHODS: Of the 51 patients with OO with PAAD enrolled in this study, 28 developed PAAD-induced myelopathy (myelopathy group), and the other 23 patients had no myelopathy (control group). Neurologic function was evaluated by the neurologic function rating system and the Japanese Orthopaedic Association score system. Three novel radiographic parameters (OP [median sagittal diameter of the spinal canal from the posteroinferior edge of the ossicle to the anterosuperior edge of the spinous process of C2]/C4 SAC [space available for spinal cord] ratio, C1 posterior inclination angle, and posterior dislocation index) were measured by lateral cervical dynamic radiography. Their correlations with neurologic function were analyzed, and their reproducibility was assessed by the intraclass correlation coefficient (ICC). In addition, receiver operating characteristic curve analysis was performed. RESULTS: A significant correlation was observed between the OP/C4 SAC ratio and the neurologic function (P < 0.01), and between the C1 posterior inclination angle and the neurologic function (P < 0.01). Furthermore, their interobserver and intraobserver reliability was excellent (ICC ≥ 0.912). Receiver operating characteristic curve analysis showed that the optimal threshold value relating to myelopathy of the OP/C4 SAC ratio and C1 posterior inclination angle was 0.93 and 20°, respectively. CONCLUSIONS: The OP/C4 SAC ratio and the C1 posterior inclination angle seem to be 2 effective and objective radiographic parameters for relating myelopathy in patients with OO with PAAD. When the OP/C4 SAC ratio is <0.93 and/or the C1 posterior inclination angle is >20°, the risk of developing myelopathy should be highly suspected in patients with OO with PAAD.
RESUMO
Incurable implant-related infection may cause catastrophic consequences due to the existence of a biofilm that resists the infiltration of host immune cells and antibiotics. Innovative approaches inspired by nanomedicine, e.g., engineering innovative multifunctional bionic coating systems on the surface of implants, are becoming increasingly attractive. Herein, 2D black phosphorus nanosheets (BPs) were loaded onto a hydroxyapatite (HA)-coated metal implant to construct a BPs@HA composite coating. With its photothermal conversion effect and in situ biomineralization, the BPs@HA coating shows excellent performances in ablating the bacterial biofilm and accelerating fracture healing, which were verified through both in vitro and in vivo studies. Moreover, differentially expressed genes of bone formation and bone mesenchymal stem cells (BMSCs) regulated by the BPs@HA coating were identified using absolute quantitative transcriptome sequencing followed by the screening of gene differential expressions. A functional enrichment analysis reveals that the expression of core markers related to BMSC differentiation and bone formation could be effectively regulated by BPs through a metabolism-related pathway. This work not only illustrates the great potential in clinical application of the BPs@HA composite coating to eliminate bacteria and accelerate bone fracture healing but also contributes to an understanding of the underlying molecular mechanism of osteogenesis physiological function regulation based on an analysis of absolute quantitative transcriptome sequencing.
Assuntos
Consolidação da Fratura , Fósforo , Fósforo/farmacologia , Durapatita/farmacologia , Osteogênese , Biofilmes , Aceleração , Antibacterianos/farmacologia , Materiais Revestidos Biocompatíveis/farmacologia , Titânio/farmacologiaRESUMO
Foot-and-mouth disease (FMD) is characterized by a pronounced lymphopenia that is associated with immune suppression. However, the mechanisms leading to lymphopenia remain unclear. In this study, the number of total CD4+, CD8+ T cells, B cells, and NK cells in the peripheral blood were dramatically reduced in C57BL/6 mice infected with foot-and-mouth disease virus (FMDV) serotype O, and it was noted that mice with severe clinical symptoms had expressively lower lymphocyte counts than mice with mild or without clinical symptoms, indicating that lymphopenia was associated with disease severity. A further analysis revealed that lymphocyte apoptosis and trafficking occurred after FMDV infection. In addition, coinhibitory molecules were upregulated in the expression of CD4+ and CD8+ T cells from FMDV-infected mice, including CTLA-4, LAG-3, 2B4, and TIGIT. Interestingly, the elevated IL-10 in the serum was correlated with the appearance of lymphopenia during FMDV infection but not IL-6, IL-2, IL-17, IL-18, IL-1ß, TNF-α, IFN-α/ß, TGF-ß, and CXCL1. Knocking out IL-10 (IL-10-/-) mice or blocking IL-10/IL-10R signaling in vivo was able to prevent lymphopenia via downregulating apoptosis, trafficking, and the coinhibitory expression of lymphocytes in the peripheral blood, which contribute to enhance the survival of mice infected with FMDV. Our findings support that blocking IL-10/IL-10R signaling may represent a novel therapeutic approach for FMD.
Assuntos
Vírus da Febre Aftosa/imunologia , Febre Aftosa/virologia , Interleucina-10/imunologia , Linfopenia/virologia , Animais , Apoptose , Linfócitos B/imunologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Citocinas/imunologia , Interleucina-10/genética , Células Matadoras Naturais/imunologia , Contagem de Linfócitos , Linfopenia/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Transdução de SinaisRESUMO
Bone defects remains a challenge for surgeons. Bone graft scaffold can fill the defect and enhance the bone regeneration. Demineralized bone matrix (DBM) is an allogeneic bone graft substitute, which can only be used as a filling material rather than a structural bone graft. Coating of the scaffolds with nanoscale DBM may enhance the osteoinductivity or osteoconductivity. Herein the lyophilization method is presented to coat the nano-DBM on surface of the porous polycaprolactone (PCL)/ß-tricalcium phosphate (ß-TCP) scaffolds fabricated by 3D printing technology. The morphology, elastic modulus, in vitro cell biocompatibility, and in vivo performance are investigated. Scanning electron microscope (SEM) shows DBM particle clusters with size of 200-500 nm are observed on scaffolds fibers after coating. MC3T3-E1 cells on nano-DBM coated PCL/ß-TCP scaffold show better activity than on PCL/ß-TCP scaffold. In vivo tests show better infiltration of new bone tissue in nano-DBM coated PCL/ß-TCP scaffold than PCL/ß-TCP scaffold via the interface. These results show the presence of nano-DBM coating on PCL/ß-TCP scaffold could enhance the attachment, proliferation, and viability of cells and benefit for the new bone formation surrounding and deep inside the scaffolds. Nano-DBM could potentially be used as a new kind of biomaterial for bone defect treatment.
Assuntos
Matriz Óssea/fisiologia , Regeneração Óssea/efeitos dos fármacos , Fosfatos de Cálcio/farmacologia , Nanopartículas/química , Poliésteres/farmacologia , Alicerces Teciduais/química , Animais , Matriz Óssea/efeitos dos fármacos , Linhagem Celular , Camundongos , Nanopartículas/ultraestrutura , Tamanho do Órgão/efeitos dos fármacos , Implantação de Prótese , Microtomografia por Raio-XRESUMO
Curcumin a component of turmeric, which is derived from Curcuma longa is used as a colouring agent and as a dietary spice for centuries. Extensive studies have been done on the anti-inflammatory activity of curcumin along with its molecular mechanism involving different signalling pathways. However, the physicochemical and biological properties such as poor solubility and rapid metabolism of curcumin have led to low bioavailability and hence limits its application. Current therapies for asthma such as bronchodilators and inhaled corticosteroids (ICS) are aimed at controlling disease symptoms and prevent asthma exacerbation. However, this approach requires lifetime therapy and is associated with a constellation of side effects. This creates a clear unmet medical need and there is an urgent demand for new and more-effective treatments. The present study is aimed to formulate liposomes containing curcumin and evaluate for its anti-inflammatory effects on lipopolysaccharide (LPS)-induced inflammation on BCi-NS1.1 cell line. Curcumin and salbutamol liposomes were formulated using lipid hydration method. The prepared liposomes were characterized in terms of particle size, zeta potential, encapsulation efficiency and in-vitro release profile. The liposomes were tested on BCI-NS1.1 cell line to evaluate its anti-inflammatory properties. The various pro-inflammatory markers studied were Interleukin-6 (IL-6), Interleukin-8 (IL-8), Interleukin-1ß (IL-1ß) and Tumour Necrosis Factor-a (TNF-a). Additionally, molecular mechanics simulations were used to elucidate the positioning, energy minimization, and aqueous dispersion of the liposomal architecture involving lecithin and curcumin. The prepared curcumin formulation showed an average size and zeta potential of 271.3 ± 3.06 nm and -61.0 mV, respectively. The drug encapsulation efficiency of liposomal curcumin is 81.1%. Both curcumin-loaded liposomes formulation (1 µg/mL, 5 µg/mL) resulted in significant (p < 0.05) reduction in the level of pro-inflammatory marker expression such as IL-6, IL-8, IL-1ß and TNF-a compared to positive control group. Liposomal curcumin with the dose of 1 µg/mL reduced the inflammatory markers more effectively compared to that of 5 µg/mL. Liposomal curcumin could be a promising intervention for asthma therapy showing their efficacy in suppressing the important pro-inflammatory markers involved in the pathogenesis of asthma.
Assuntos
Asma/tratamento farmacológico , Curcumina/uso terapêutico , Asma/patologia , Biomarcadores/metabolismo , Linhagem Celular , Simulação por Computador , Curcumina/química , Liberação Controlada de Fármacos , Humanos , Inflamação/patologia , Lipossomos , Modelos Moleculares , Tamanho da Partícula , Espectroscopia de Infravermelho com Transformada de Fourier , Eletricidade EstáticaRESUMO
Several vesicular systems loaded with curcumin have found their way in the therapeutic applications of several diseases, primarily acting through their immunological pathways. Such systems use particles at a nanoscale range, bringing about their intended use through a range of complex mechanisms. Apart from delivering drug substances into target tissues, these vesicular systems also effectively overcome problems like insolubility and unequal drug distribution. Several mechanisms are explored lately by different workers, and interest over vesicular curcumin has been renewed in the past decade. This commentary discusses several immunological targets in which curcumin is employed in a vesicular form.
Assuntos
Analgésicos/administração & dosagem , Anti-Inflamatórios/administração & dosagem , Antineoplásicos/administração & dosagem , Curcumina/administração & dosagem , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos , Humanos , Lipossomos/química , Nanopartículas/química , SolubilidadeRESUMO
OBJECTIVE: To explore the effect of long-term stress on the temporomandibular joint (TMJ) condyle and its possible underlying mechanism. METHODS: A 12-week, chronic unpredictable mild stress (CUMS) model was used to induce long-term psychological stress in rats. Rats were randomly divided into control group (CONT), chronic unpredictable mild stress group (CUMS) and chronic unpredictable mild stress with fluoxetine treatment group (CUMS + DT) (n = 30 per group). A 5 mg/kg dose of fluoxetine was intraperitoneally injected daily 0.5 h before stress. A sucrose preference test, plasma corticosterone test and open-field test were performed to verify the feasibility of the CUMS model. Histopathology was used to observe the pathological changes of condyle. The expression levels of inflammatory cytokines, matrix metalloproteases (MMPs) and extracellular matrix (ECM) were measured by real-time polymerase chain reaction, western blotting and immunohistochemistry. RESULTS: At 8 and 12 weeks after exposure to CUMS, the rats showed higher plasma corticosterone than the control rats. Additionally, for the open-field test, the rats exposed to CUMS spent more time in the centre zone and moved a shorter distance than the control and drug treatment rats. In addition, pathological changes in the condylar cartilage occurred in the 8-week CUMS subgroup and were more obvious in the 12-week CUMS subgroup. The CUMS caused an increase in the secretion of inflammatory cytokines, imbalanced expression of MMPs and tissue inhibitor of metalloproteinase-1 and accelerated degradation of ECM in condylar cartilage in a time-dependent manner. CONCLUSION: Osteoarthritis-like lesions can be caused by long-term CUMS in the mandibular condyles, which suggests that the imbalance in chondrocyte-secreted regulatory factors within the cartilage of the TMJ may play an important role in cartilage injury induced by psychological stress.
Assuntos
Cartilagem Articular/metabolismo , Matriz Extracelular/metabolismo , Côndilo Mandibular/metabolismo , Estresse Psicológico/metabolismo , Agrecanas/efeitos dos fármacos , Agrecanas/genética , Agrecanas/metabolismo , Animais , Comportamento Animal , Western Blotting , Cartilagem Articular/efeitos dos fármacos , Colágeno Tipo II/efeitos dos fármacos , Colágeno Tipo II/genética , Colágeno Tipo II/metabolismo , Corticosterona/metabolismo , Citocinas/metabolismo , Ensaio de Imunoadsorção Enzimática , Matriz Extracelular/efeitos dos fármacos , Fluoxetina/farmacologia , Imuno-Histoquímica , Masculino , Côndilo Mandibular/efeitos dos fármacos , Metaloproteinase 3 da Matriz/efeitos dos fármacos , Metaloproteinase 3 da Matriz/genética , Metaloproteinase 3 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/efeitos dos fármacos , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Inibidores Seletivos de Recaptação de Serotonina/farmacologia , Estresse Psicológico/genética , Articulação Temporomandibular , Inibidor Tecidual de Metaloproteinase-1/efeitos dos fármacos , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/metabolismoRESUMO
BACKGROUND: Vesicular systems like nanotechnology and liposomes are gaining tremendous attention lately in the field of respiratory diseases. These formulations enhance bioavailability of the drug candidate, which could be achieved through a novel drug delivery mechanism. Moreover, the therapeutic potential achieved through these systems is highly controllable over long durations of time providing better efficacy and patient compliance. OBJECTIVE: The objective of this paper is to review the recent literature on vesicular drug delivery systems containing curcumin. METHODS: We have collated and summarized various recent attempts made to develop different controlled release drug delivery systems containing curcumin which would be of great interest for herbal, formulation and biological scientists. There are several vesicular nanotechnological techniques involving curcumin which have been studied recently, targeting pulmonary diseases. RESULTS: Different vesicular systems containing curcumin are being studied for their therapeutic potential in different respiratory diseases. There has been a renewed interest in formulations containing curcumin recently, primarily owing to the broad spectrum therapeutic potential of this miracle substance. Various types of formulations, containing curcumin, targeting different bodily systems have recently emerged and, nevertheless, the search for newer frontiers with this drug goes on. CONCLUSION: This mini review, in this direction, tries to highlight the key research interventions employing vesicular systems of drug delivery with curcumin.
Assuntos
Curcumina/química , Sistemas de Liberação de Medicamentos/métodos , Transtornos Respiratórios/tratamento farmacológico , Asma/tratamento farmacológico , Asma/fisiopatologia , Curcumina/administração & dosagem , Curcumina/farmacologia , Preparações de Ação Retardada , Composição de Medicamentos/métodos , Humanos , Lipossomos/química , Nanomedicina/métodos , Nanopartículas/química , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Doença Pulmonar Obstrutiva Crônica/imunologiaRESUMO
Cellulose isolation, a promising way for lignocellulosic biomass utilization, is always restricted by the poor solubility of cellulose. In this paper, tetra-n-methylammonium hydroxide (TMAH) was confirmed to be capable of readily dissolving/regenerating cellulose without chemical modification at room temperature. Meanwhile, cellulose isolation from lignocellulosic biomass by initially dissolving the biomass in TMAH followed by cellulose precipitation was proposed, and the isolated substance with average cellulose purity of 92.1 ± 0.3% could be obtained throughout this process under the optimum conditions: temperature 52 °C, time 60 min, and loading ratio of TMAH/biomass (w/w) 7.2:1. Besides, high efficiency cellulose isolation (i.e. >70% cellulose purity) could be continuously remained during 4-round cycles by using the recycled TMAH solvent without distinct activity loss.
Assuntos
Celulose/química , Compostos de Amônio Quaternário/químicaRESUMO
The aim of this study was to construct a cell transplant method consisting of cell sheet fragments of periodontal ligament stem cells (PDLSCs) and platelet-rich fibrin (PRF) granules to enhance periodontal healing in avulsed tooth reimplantation. To test this concept in vitro, human PDLSCs were isolated and characterized by colony forming unit assay, cell surface marker characterizations, and their osteogenic/adipogenic differentiation potential. The biological effects of autologous PRF as a growth factor-enriched endogenous scaffold on human PDLSCs were then investigated and quantified for statistical analyses, including cell viability and proliferation, alkaline phosphatase (ALP) activity, and the gene expression of bone sialoprotein (BSP), osteocalcin (OCN), collagen I (Col-I), and cementum protein 23 (CP23). It was found that the PRF induced a significant and continuous stimulation of proliferation in human PDLSCs throughout the 7-day incubation period. Furthermore, the PRF suppressed the osteoblastic differentiation of PDLSCs by decreasing ALP activity and the gene expression of BSP and OCN while up-regulating the mRNA expression levels of Col-I and CP23 during the testing period. To assess the potential application of the PDLSCs/PRF construct in tooth reimplantation, 36 incisors were extracted from 6 dogs. The incisors then underwent 2 h of dry storage and were randomly divided into four groups receiving different strategies of reimplantation, where the avulsed teeth were reimplanted with the use of the autologous PDLSCs/PRF construct (cell sheet fragments in combination with PRF granules), with the use of autologous PDLSCs or PRF alone, or without adjuvant use of PRF or PDLSCs. Eight weeks post-reimplantation, the PDLSCs/PRF group achieved a more effective periodontal healing, characterized by the regeneration of PDL-like tissues and a reduction of ankylosis and inflammation, compared with the other testing groups. These overall results suggest that the PDLSCs/PRF construct may be a useful tool for alveolar surgery that has the potential to improve the clinical outcomes in future avulsed tooth reimplantations.
Assuntos
Plaquetas/química , Técnicas de Cultura de Células/métodos , Fibrina/farmacologia , Ligamento Periodontal/citologia , Células-Tronco/citologia , Avulsão Dentária/terapia , Reimplante Dentário , Adulto , Fosfatase Alcalina/metabolismo , Animais , Plaquetas/ultraestrutura , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Proliferação de Células/efeitos dos fármacos , Separação Celular , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/genética , Células Cultivadas , Cães , Fibrina/ultraestrutura , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Contagem de Plaquetas , Transplante de Células-Tronco , Células-Tronco/efeitos dos fármacos , Células-Tronco/enzimologia , Células-Tronco/ultraestrutura , Cicatrização/efeitos dos fármacos , Adulto JovemRESUMO
In the water-in-[Bmim][PF(6)] microemulsion stabilized by both AOT and Triton X-100, the lipase-catalyzed hydrolysis of 4-nitrophenyl butyrate (p-NPB) was investigated to evaluate the catalytic efficiency of lipase in this novel microemulsion. The structural parameters of the microemulsion and the conditions of the enzymatic reaction affect the catalytic activity of lipase, especially the concentration of Tris-HCl buffer. Under optimum conditions, the catalytic activity of lipase in the present microemulsion is much higher than that in H(2)O saturated [Bmim][PF(6)]. When the partitioning of the substrate in the microemulsion is taken into account, the catalytic efficiency of lipase in this novel microemulsion is 14.3 times that in H(2)O saturated [Bmim][PF(6)] due to the significant decrease of the Michaelis constant in the microemulsion. Due to the large interface, high water activity, and probably the activating effect of the imidazolium cation in the water pool, the present microemulsion is demonstrated to be a promising medium for the lipase-catalyzed hydrolytic reaction. To demonstrate an important biocatalytic application in the IL-based microemulsion, the lipase-catalyzed synthesis of the flavoring agent benzyl acetate via transesterification of vinyl acetate with benzyl alcohol was also studied in the medium. Due to the high dispersion of lipase, large interface and removal of the byproduct, a maximum yield of 94% was obtained, indicating that the novel microemulsion is really important and useful.
Assuntos
Biocatálise/efeitos dos fármacos , Imidazóis/farmacologia , Lipase/metabolismo , Octoxinol/farmacologia , Succinatos/farmacologia , Água/química , Absorção/efeitos dos fármacos , Compostos de Benzil/metabolismo , Soluções Tampão , Butiratos/metabolismo , Candida/enzimologia , Emulsões , Concentração de Íons de Hidrogênio/efeitos dos fármacos , Hidrólise/efeitos dos fármacos , Cinética , Análise Espectral , Tensão Superficial/efeitos dos fármacos , Tensoativos/farmacologia , Fatores de TempoRESUMO
OBJECTIVE: To identify enterovirus 71 (EV71) strains isolated from patients with hand, foot and mouth disease (HFMD) in Guangdong and Fujian provinces from 2000 to 2001 by using phylogenetic analysis. METHODS: All 25 samples were first tested for enteric viruses by RT-PCR using enterovirus specific primers EV-1 and EV-2, and then were identified for EV71 by RT-PCR using EV71 specific primers 159S and 162A. The amplicons of 485bp segment (part of the VP1 gene) were cloned into pGEM-T and sequenced. A phylogenetic tree was constructed by comparison of the sequences with other 12 EV71 strains isolated from China, Japan, Hungary, and the United States including the prototype BrCr. RESULTS: The positive rate of EV71 was about 20%. The sequence analysis showed that the new isolate (GZH2000) shared 94%-96% nucleotide identity with three strains isolated in 1998 and 2000, and 91% with a strain isolated in 1987 from Chinese mainland, but shared only 82%-84% homology with EV71 isolates studied abroad. CONCLUSIONS: EV71 is one of the important pathogens of HFMD in south China. The strains isolated from mainland were closely related with most isolates from Taiwan, but different from most EV71 strains reported abroad. The symptoms of EV71 infection in mainland were not as intensive as those described in Taiwan's outbreak.
Assuntos
Enterovirus/genética , Doença de Mão, Pé e Boca/virologia , Filogenia , Adolescente , Criança , Pré-Escolar , China , Enterovirus/isolamento & purificação , Humanos , Lactente , Análise de Sequência , Homologia de Sequência do Ácido NucleicoRESUMO
OBJECTIVE: To investigate the epidemiological characteristus of human caliciviruses (HuCVs) among children under 5 years of age with acute diarrhea and to estimate the disease burden in Lulong county. METHODS: HuCVs were detected by enzyme linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). Some PCR amplicons were cloned and sequenced. Phylogenetic tree was constructed for strain characterization. The rate of HuCVs-attributed hospitalization was estimated according to the positive rate of HuCVs detection in fecal specimens collected from hospitalized diarrhea patients. RESULTS: Between July 1999 and June 2001, 708 fecal specimens were collected, of which 393 rotavirus-negative and 5 rotavirus-positive specimens were detected for HuCVs. Thirty-one point six percentage of fecal specimens from patients with diarrhea was HuCVs positive. Among inpatients, HuCVs positive rate was 17.5%. HuCVs detection was mainly distributed in 3 - 17 mouth-old children, in winter. All 11 strains belonged to NLV GII in which 6 strains GII-3, 2 strains GII-4 and 3 strains GII-7, and they shared 55.1% - 100% nucleotide identity. NLV GII-4 and GII-7 were identified in 2000, while NLV GII-3 and GII-7 in 2001. The preliminary estimate of HuCVs-attributed hospitalization rate was 3.6 per thousand. CONCLUSION: Human caliciviruses with different genotypes circulated among children in Lulong county with GII NLVs were the prevalent strains. The disease burden of HuCVs was second to rotavirus.