RESUMEN
Introducing component-selective polymer chains onto the surface of a particle is an effective approach to improve the compatibilization efficiency of a particle-based compatibilizer. In this study, two particles with different kinds of component-selective polymer chains that have the same length and similar density but different graft locations were synthesized and their compatibilization effects were comparatively investigated. It was found that compared with the particle with homogeneous PMMA and PP grafts (R-P), the particle with a hemisphere of poly(methyl methacrylate) (PMMA) grafts and other hemisphere of polypropylene (PP) chains (J-P) showed a better compatibilization effect under equal loadings, although both particles exhibited high efficiency. The better compatibilization effect of particles with Janus grafts may be attributed to the stronger entanglements between grafted polymer chains and selective individual components. This work suggests that optimizing the graft location of a particle is an effective strategy for improving its compatibilization efficiency and helpful for the design of advanced particle compatibilizers.
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A facile covalent assembly strategy is proposed for the preparation of superhydrophobic COF-stabilized MXene separation membranes. Ultra-high separation fluxes of up to 54 280 L m-2 h-1 and 643 200 L m-2 h-1 bar-1 are obtained for emulsified water-in-oil mixtures by adopting gravity and external pressure, respectively. Moreover, the challenges of easy swelling and oxidation properties of MXene have been effectively overcome via the COF-stabilized mechanism.
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Selective hydrogenation of alkynes to obtain alkenes is a key reaction in petrochemical and fine chemical industries. However, the development of stable and highly selective catalysts with uniformly dispersed active sites is still immensely challenging for the semi-hydrogenation of alkynes. In this study, N-doped porous carbon nanospheres (NPCNs) were synthesized by the nanoemulsion self-assembly and subsequently carbonization method. Ultrafine PdCu bimetallic nanoparticles (NPs) were uniformly dispersed and immobilized on NPCNs. The obtained PdCu/NPCNs catalyst exhibited an open framework and abundant active sites originating from ultrafine PdCu NPs. In the semi-hydrogenation of alkynes, the PdCu/NPCNs catalyst exhibited a remarkable performance and stability, outperforming most of the classical catalysts. The excellent performance was related to the introduction of a secondary metal Cu, which can regulate the electronic state of Pd active sites to further enhance the hydrogenation activity and selectivity. Hence, the facile approach reported herein may be useful for constructing highly dispersed bimetallic NP-based catalysts for selective hydrogenation of alkynes in the petrochemical industry.
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Highly transparent, durable, flexible and smooth coatings with excellent anti-fouling properties have broad applications on cars, windows, and touch screens. However, the coexistence of these multi-functions is difficult to achieve in a single coating material. Here, a coating is developed with excellent performance of high transparency (98.8%), anti-fouling, high hardness (8H), and flexibility simultaneously (TAHF coating). In the material design, methyl etherified melamine formaldehyde resin, hexamethylene diisocyanate trimer, and mono-aminopropyl terminated polydimethylsiloxane (NH2-PDMS) were used as a polymer matrix to provide surface hardness, a cross-linker was used to provide toughness, and omniphobic groups from NH2-PDMS were used to provide anti-fouling performance. The TAHF coating has excellent liquid repellence even after six months of outdoor exposure, 260 h of UV light exposure, and 1500 wear and 2000 bending cycles, and its chemical shielding performance is superior to that of a commercial anti-corrosive coating. This strategy would provide a new route for the design of multifunctional anti-fouling coatings for practical applications.
RESUMEN
Chemoselective hydrogenation of unsaturated organic compounds is a significant research topic in the catalysis field. Herein, a sulfur-doped ordered mesoporous carbon (SMC) material was prepared to anchor ultrafine platinum (Pt) clusters for the chemoselective hydrogenation of halogenated nitroarenes. The confinement effect of the ordered pores and the strong metal-support interaction caused by Pt clusters and sulfur atoms, efficiently suppress the aggregation and regulate the electronic states of the ultrafine Pt clusters. Thus, the hydrogenation of parachloronitrobenzene (p-CNB) shows high selectivity catalyzed by the ultrafine Pt clusters with electron-rich states. Meanwhile, the catalytic performance of the hydrogenation reaction catalyzed by Pt/SMC is capable of being maintained after at least 5 cycles, and the catalytic universality can also be applied to different halogenated nitroarenes hydrogenation. Therefore, this study may promote the research into the construction of noble metal-based catalysts for chemoselective hydrogenation reactions in green and sustainable chemical processes.
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The goals of a genital herpes vaccine are to prevent painful genital lesions and reduce or eliminate subclinical infection that risks transmission to partners and newborns. We evaluated a trivalent glycoprotein vaccine containing herpes simplex virus type 2 (HSV-2) entry molecule glycoprotein D (gD2) and two immune evasion molecules: glycoprotein C (gC2), which binds complement C3b, and glycoprotein E (gE2), which blocks immunoglobulin G (IgG) Fc activities. The trivalent vaccine was administered as baculovirus proteins with CpG and alum, or the identical amino acids were expressed using nucleoside-modified mRNA in lipid nanoparticles (LNPs). Both formulations completely prevented genital lesions in mice and guinea pigs. Differences emerged when evaluating subclinical infection. The trivalent protein vaccine prevented dorsal root ganglia infection, and day 2 and 4 vaginal cultures were negative in 23 of 30 (73%) mice compared with 63 of 64 (98%) in the mRNA group (P = 0.0012). In guinea pigs, 5 of 10 (50%) animals in the trivalent subunit protein group had vaginal shedding of HSV-2 DNA on 19 of 210 (9%) days compared with 2 of 10 (20%) animals in the mRNA group that shed HSV-2 DNA on 5 of 210 (2%) days (P = 0.0052). The trivalent mRNA vaccine was superior to trivalent proteins in stimulating ELISA IgG antibodies, neutralizing antibodies, antibodies that bind to crucial gD2 epitopes involved in entry and cell-to-cell spread, CD4+ T cell responses, and T follicular helper and germinal center B cell responses. The trivalent nucleoside-modified mRNA-LNP vaccine is a promising candidate for human trials.
Asunto(s)
Herpes Genital/inmunología , Nucleósidos/inmunología , ARN Mensajero/inmunología , Proteínas del Envoltorio Viral/inmunología , Animales , Cobayas , ARN Mensajero/biosíntesis , Proteínas del Envoltorio Viral/biosíntesisRESUMEN
The direct oxidation of alcohols to esters through a green and cost-effective strategy is a fascinating chemical synthesis route. In this study, an environmentally friendly N-doped porous carbon encapsulated Co-based nano-catalyst was prepared via a simple carbonization procedure, utilizing renewable chitosan, accessible dicyandiamide and low-cost Co(OAc)2 as co-precursors. The obtained Co@NC-2-T catalysts were successfully used in selective oxidation of aromatic alcohols with methanol to esters under atmospheric reaction conditions. The Co@NC-2-900 catalyst (added with 2 g dicyandiamide and pyrolyzed at 900 °C) shows optimal activity and applicability and can also be reused at least six times in the oxidative esterification of aromatic alcohols with excellent stability. The presence of superoxide anion radicals in the current catalytic system was detected by the EPR method, and a possible mechanism of alcohol oxidation to ester was proposed on this basis. Thus, this study provides a facile, eco-friendly, and highly efficient catalytic system for oxidative esterification of alcohols.
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Alcoholes/química , Carbono/química , Cobalto/química , Nitrógeno/química , Catálisis , Esterificación , Oxidación-Reducción , PorosidadRESUMEN
The fabrication of ultrafine noble metal nanoparticle (NMNP)-based catalysts is significant for heterogeneous catalysis due to their excellent performance for organic transformations. In this study, N-doped micro-mesoporous hollow carbon nanospheres (HCN) with a nitrogen content of â¼3.5â¯wt% are easily prepared by simple carbonization. Then, ultrafine Pd NPs are immobilized on HCN, affording Pd/HCN as a dual-function catalyst which exhibits superior catalytic activity for the selective oxidation of 5-hydroxymethylfurfural (HMF) and hydrogenation of nitroarenes, under mild reaction conditions. The doping of HCN with nitrogen is beneficial for the high dispersion, anchoring, and particle size control of ultrafine Pd NPs, leading to the maximum utilization of Pd atoms and further enhancing the catalytic activity of Pd/HCN. In addition, the obtained Pd/HCN catalyst exhibits excellent reusability and stability. Hence, this study demonstrates the prospect of developing ultrafine NMNP-supported catalysts with high performance for organic transformations.
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The design of lower-cost alternative heterogeneous catalysts for the hydrogenation of halogenated nitrobenzenes using green method to synthesize the corresponding anilines is highly desirable. In this study, Ag was incorporated into the Co-MOFs during the growing process (Co-Ag(n)-MOFs), and then followed the carbothermal reduction process without any additional procedures, we synthesized a series of Co-Ag(n)@NCs. The self-supported catalysts exhibited excellent and stable catalytic performances for the chemoselective hydrogenation of halogenated nitrobenzenes without obvious dehalogenation. The Co-Ag bimetallic alloy nanoparticles were well-dispersed and protected from aggregation and leaching by the porous nitrogen doped carbon. Besides, either hydrazine hydrate (N2H4·H2O, generating byproducts N2 and H2O) or H2 could be used as green reducing agent with excellent selectivity towards synthesizing the corresponding anilines. And when the Co/Ag content ratio was approximate 1:1, the Co-Ag(1:1)@NC showed the best catalytic performance. Moreover, the Co-Ag(1:1)@NC could be efficiently recovered by using an external magnetic force and reused without obvious decrease of catalytic activity. Thus, such highly efficient, inexpensive, stable and magnetically recyclable catalysts could show great potentials in practical applications for many important reactions.
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A herpes simplex virus 2 (HSV-2) glycoprotein E deletion mutant (gE2-del virus) was evaluated as a replication-competent, attenuated live virus vaccine candidate. The gE2-del virus is defective in epithelial cell-to-axon spread and in anterograde transport from the neuron cell body to the axon terminus. In BALB/c and SCID mice, the gE2-del virus caused no death or disease after vaginal, intravascular, or intramuscular inoculation and was 5 orders of magnitude less virulent than wild-type virus when inoculated directly into the brain. No infectious gE2-del virus was recovered from dorsal root ganglia (DRG) after multiple routes of inoculation; however, gE2-del DNA was detected by PCR in lumbosacral DRG at a low copy number in some mice. Importantly, no recurrent vaginal shedding of gE2-del DNA was detected in immunized guinea pigs. Intramuscular immunization outperformed subcutaneous immunization in all parameters evaluated, although individual differences were not significant, and two intramuscular immunizations were more protective than one. Immunized animals had reduced vaginal disease, vaginal titers, DRG infection, recurrent genital lesions, and recurrent vaginal shedding of HSV-2 DNA; however, protection was incomplete. A combined modality immunization using live virus and HSV-2 glycoprotein C and D subunit antigens in guinea pigs did not totally eliminate recurrent lesions or recurrent vaginal shedding of HSV-2 DNA. The gE2-del virus used as an immunotherapeutic vaccine in previously HSV-2-infected guinea pigs greatly reduced the frequency of recurrent genital lesions. Therefore, the gE2-del virus is safe, other than when injected at high titer into the brain, and is efficacious as a prophylactic and immunotherapeutic vaccine.
Asunto(s)
Eliminación de Gen , Vacunas contra el Virus del Herpes Simple/inmunología , Herpesvirus Humano 2/genética , Herpesvirus Humano 2/inmunología , Neuronas/virología , Proteínas del Envoltorio Viral/genética , Proteínas del Envoltorio Viral/inmunología , Animales , Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/inmunología , Línea Celular , Chlorocebus aethiops , ADN Viral , Femenino , Ganglios Espinales/virología , Cobayas , Herpes Genital/mortalidad , Herpes Genital/prevención & control , Herpes Genital/terapia , Herpes Simple/mortalidad , Herpes Simple/prevención & control , Herpes Simple/terapia , Vacunas contra el Virus del Herpes Simple/administración & dosificación , Vacunas contra el Virus del Herpes Simple/efectos adversos , Ratones , Ratones Endogámicos BALB C , Ratones SCID , Médula Espinal/virología , Vacunas Atenuadas/administración & dosificación , Vacunas Atenuadas/efectos adversos , Vacunas Atenuadas/inmunologíaRESUMEN
Attempts to develop a vaccine to prevent genital herpes simplex virus 2 (HSV-2) disease have been only marginally successful, suggesting that novel strategies are needed. Immunization with HSV-2 glycoprotein C (gC-2) and gD-2 was evaluated in mice and guinea pigs to determine whether adding gC-2 to a gD-2 subunit vaccine would improve protection by producing antibodies that block gC-2 immune evasion from complement. Antibodies produced by gC-2 immunization blocked the interaction between gC-2 and complement C3b, and passive transfer of gC-2 antibody protected complement-intact mice but not C3 knockout mice against HSV-2 challenge, indicating that gC-2 antibody is effective, at least in part, because it prevents HSV-2 evasion from complement. Immunization with gC-2 also produced neutralizing antibodies that were active in the absence of complement; however, the neutralizing titers were higher when complement was present, with the highest titers in animals immunized with both antigens. Animals immunized with the gC-2-plus-gD-2 combination had robust CD4+ T-cell responses to each immunogen. Multiple disease parameters were evaluated in mice and guinea pigs immunized with gC-2 alone, gD-2 alone, or both antigens. In general, gD-2 outperformed gC-2; however, the gC-2-plus-gD-2 combination outperformed gD-2 alone, particularly in protecting dorsal root ganglia in mice and reducing recurrent vaginal shedding of HSV-2 DNA in guinea pigs. Therefore, the gC-2 subunit antigen enhances a gD-2 subunit vaccine by stimulating a CD4+ T-cell response, by producing neutralizing antibodies that are effective in the absence and presence of complement, and by blocking immune evasion domains that inhibit complement activation.
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Ganglios Espinales/inmunología , Herpes Genital/prevención & control , Vacuna contra el Herpes Zóster/inmunología , Herpes Zóster/prevención & control , Herpesvirus Humano 2/inmunología , Proteínas del Envoltorio Viral/inmunología , Animales , Anticuerpos Neutralizantes/sangre , Anticuerpos Antivirales/sangre , Linfocitos T CD4-Positivos/inmunología , Femenino , Cobayas , Herpes Genital/inmunología , Herpes Zóster/inmunología , Vacuna contra el Herpes Zóster/administración & dosificación , Inmunización/métodos , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Prevención Secundaria , Vagina/virología , Esparcimiento de VirusRESUMEN
Herpes simplex virus 1 (HSV-1) glycoprotein E (gE) mediates cell-to-cell spread and functions as an IgG Fc receptor (FcγR) that blocks the Fc domain of antibody targeting the virus or infected cell. Efforts to assess the functions of the HSV-1 FcγR in vivo have been hampered by difficulties in preparing an FcγR-negative strain that is relatively intact for spread. Here we report the FcγR and spread phenotypes of NS-gE264, which is a mutant strain that has four amino acids inserted after gE residue 264. The virus is defective in IgG Fc binding yet causes zosteriform disease in the mouse flank model that is only minimally reduced compared with wild-type and the rescue strains. The presence of zosteriform disease suggests that NS-gE264 spread functions are well maintained. The HSV-1 FcγR binds the Fc domain of human, but not murine IgG; therefore, to assess FcγR functions in vivo, mice were passively immunized with human IgG antibody to HSV. When antibody was inoculated intraperitoneally 20 h prior to infection or shortly after virus reached the dorsal root ganglia, disease severity was significantly reduced in mice infected with NS-gE264, but not in mice infected with wild-type or rescue virus. Studies of C3 knockout mice and natural killer cell-depleted mice demonstrated that the HSV-1 FcγR blocked both IgG Fc-mediated complement activation and antibody-dependent cellular cytotoxicity. Therefore, the HSV-1 FcγR promotes immune evasion from IgG Fc-mediated activities and likely contributes to virulence at times when antibody is present, such as during recurrent infections.
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Anticuerpos Antivirales/inmunología , Citotoxicidad Celular Dependiente de Anticuerpos , Activación de Complemento , Herpesvirus Humano 1/inmunología , Herpesvirus Humano 1/patogenicidad , Evasión Inmune , Proteínas del Envoltorio Viral/inmunología , Animales , Complemento C3/deficiencia , Complemento C3/inmunología , Modelos Animales de Enfermedad , Herpes Zóster/patología , Herpes Zóster/virología , Humanos , Células Asesinas Naturales/inmunología , Procedimientos de Reducción del Leucocitos , Ratones , Ratones Endogámicos BALB C , Índice de Severidad de la Enfermedad , Proteínas del Envoltorio Viral/genética , Proteínas del Envoltorio Viral/metabolismo , Factores de Virulencia/genética , Factores de Virulencia/inmunología , Factores de Virulencia/metabolismoRESUMEN
The HSV-2 lifecycle involves virus spread in a circuit from the inoculation site to dorsal root ganglia and return. We evaluated the role of gE-2 in the virus lifecycle by deleting amino acids 124-495 (gE2-del virus). In the mouse retina infection model, gE2-del virus does not spread to nuclei in the brain, indicating a defect in anterograde (pre-synaptic to post-synaptic neurons) and retrograde (post-synaptic to pre-synaptic neurons) spread. Infection of neuronal cells in vitro demonstrates that gE-2 is required for targeting viral proteins from neuron cell bodies into axons, and for efficient virus spread from epithelial cells to axons. The mouse flank model confirms that gE2-del virus is defective in spread from epithelial cells to neurons. Therefore, we defined two steps in the virus lifecycle that involve gE-2, including efficient spread from epithelial cells to axons and targeting viral components from neuron cell bodies into axons.
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Axones/metabolismo , Células Epiteliales/virología , Herpesvirus Humano 2/patogenicidad , Neuronas/virología , Proteínas del Envoltorio Viral/metabolismo , Proteínas Virales/metabolismo , Animales , Células Cultivadas , Chlorocebus aethiops , Modelos Animales de Enfermedad , Femenino , Herpes Simple/fisiopatología , Herpes Simple/virología , Herpesvirus Humano 2/genética , Herpesvirus Humano 2/metabolismo , Humanos , Queratinocitos/virología , Ratones , Ratones Endogámicos BALB C , Neuronas/metabolismo , Ratas , Ratas Sprague-Dawley , Retina/virología , Eliminación de Secuencia , Vagina/virología , Células Vero , Proteínas del Envoltorio Viral/química , Proteínas del Envoltorio Viral/genética , Proteínas Virales/genéticaRESUMEN
Herpes simplex virus type 1 (HSV-1) produces oral lesions, encephalitis, keratitis, and severe infections in the immunocompromised host. HSV-1 is almost as common as HSV-2 in causing first episodes of genital herpes, a disease that is associated with an increased risk of human immunodeficiency virus acquisition and transmission. No approved vaccines are currently available to protect against HSV-1 or HSV-2 infection. We developed a novel HSV vaccine strategy that uses a replication-competent strain of HSV-1, NS-gEnull, which has a defect in anterograde and retrograde directional spread and cell-to-cell spread. Following scratch inoculation on the mouse flank, NS-gEnull replicated at the site of inoculation without causing disease. Importantly, the vaccine strain was not isolated from dorsal root ganglia (DRG). We used the flank model to challenge vaccinated mice and demonstrated that NS-gEnull was highly protective against wild-type HSV-1. The challenge virus replicated to low titers at the site of inoculation; therefore, the vaccine strain did not provide sterilizing immunity. Nevertheless, challenge by HSV-1 or HSV-2 resulted in less-severe disease at the inoculation site, and vaccinated mice were totally protected against zosteriform disease and death. After HSV-1 challenge, latent virus was recovered by DRG explant cocultures from <10% of vaccinated mice compared with 100% of mock-vaccinated mice. The vaccine provided protection against disease and death after intravaginal challenge and markedly lowered the titers of the challenge virus in the vagina. Therefore, the HSV-1 gEnull strain is an excellent candidate for further vaccine development.
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Virus Defectuosos/inmunología , Vacunas contra el Virus del Herpes Simple/inmunología , Herpesvirus Humano 1/inmunología , Herpesvirus Humano 1/fisiología , Replicación Viral/fisiología , Adolescente , Animales , Niño , Femenino , Vacunas contra el Virus del Herpes Simple/efectos adversos , Herpesvirus Humano 1/aislamiento & purificación , Humanos , Inmunización Secundaria/métodos , Ratones , Ratones Endogámicos BALB C , Vacunas Atenuadas/inmunologíaRESUMEN
Herpes simplex virus type 1 (HSV-1) glycoprotein E (gE) promotes cell-to-cell spread at basolateral surfaces of epithelial cells, but its activity in neurons is less clear. We used the mouse retina infection model and neuronal cell cultures to define the spread phenotype of gE mutant viruses. Wild-type (WT) and gE-null (NS-gEnull) viruses both infected retina ganglion cell neurons; however, NS-gEnull viral antigens failed to reach the optic nerve, which indicates a defect in axonal localization. We evaluated two Fc receptor-negative gE mutant viruses containing four amino acid inserts in the gE ectodomain. One mutant virus failed to spread from the retina into the optic nerve, while the other spread normally. Therefore, the gE ectodomain is involved in axonal localization, and the Fc receptor and neuronal spread are mediated by overlapping but distinct gE domains. In the retina infection model, virus can travel to the brain via the optic nerve from presynaptic to postsynaptic neurons (anterograde direction) or via nerves that innervate the iris and ciliary body from postsynaptic to presynaptic neurons (retrograde direction). WT virus infected the brain by anterograde and retrograde routes, whereas NS-gEnull virus failed to travel by either pathway. The site of the defect in retrograde spread remains to be determined; however, infection of rat superior cervical ganglia neurons in vitro indicates that gE is required to target virion components to the axon initial segment. The requirement for gE in axonal targeting and retrograde spread highlights intriguing similarities and differences between HSV-1 and pseudorabies virus gE.
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Transporte Axonal , Cápside/metabolismo , Herpes Simple/virología , Simplexvirus/fisiología , Proteínas del Envoltorio Viral/metabolismo , Proteínas del Envoltorio Viral/fisiología , Proteínas Estructurales Virales/metabolismo , Animales , Encéfalo/virología , Células Cultivadas , Modelos Animales de Enfermedad , Ratones , Mutación , Nervio Óptico/virología , Ratas , Receptores Fc/genética , Retina/virología , Simplexvirus/metabolismo , Simplexvirus/patogenicidad , Ganglio Cervical Superior/virología , Proteínas del Envoltorio Viral/genética , Virulencia , Replicación ViralRESUMEN
Claudins are proteins that participate in epithelial barrier function and regulate paracellular permeability. By immunohistochemistry of adult rat lung sections, claudin-3, claudin-4, and claudin-5 were found to be co-expressed by type II alveolar epithelial cells. Claudin-3 and claudin-4 were also co-expressed by some alveolar epithelial cells adjacent to type II cells. In contrast, claudin-5 was expressed throughout the alveolus. Isolated primary rat alveolar epithelial cells in culture also expressed claudin-3, claudin-4, and claudin-5, but showed little claudin-1 and claudin-2 expression. Claudin expression by isolated cells at both the mRNA and protein level varied with time in culture. In particular, claudin-3 and claudin-5 co-localized and were distributed around the alveolar cell periphery, but claudin-4 expression was heterogeneous. We also found that paracellular permeability was increased when cultured alveolar epithelial cells were treated with a fatty acid amide, methanandamide. Methanandamide did not alter cell viability. Claudin-3, claudin-4, claudin-5, occludin, and zona occludens 1 remained localized to cell-cell contact sites at the plasma membrane in methanandamide-treated cells, suggesting that plasma membrane localization of these junction proteins is not sufficient for maintaining barrier function. However, methanandamide-treated cells showed a 12-fold increase in claudin-5 expression and a 2- to 3-fold increase in claudin-3, consistent with the notion that specific changes in claudin expression levels may correlate with changes in alveolar epithelial barrier function.
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Células Epiteliales/metabolismo , Proteínas de la Membrana/metabolismo , Alveolos Pulmonares/metabolismo , Transportadoras de Casetes de Unión a ATP/metabolismo , Animales , Ácidos Araquidónicos/farmacología , Permeabilidad de la Membrana Celular/efectos de los fármacos , Células Cultivadas , Claudina-1 , Claudina-3 , Claudina-4 , Claudina-5 , Claudinas , Células Epiteliales/efectos de los fármacos , Proteínas de la Membrana/efectos de los fármacos , Proteínas de la Membrana/genética , Microscopía Fluorescente , Alveolos Pulmonares/citología , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-Dawley , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
To define further the mechanisms of gap junction protein (connexin (Cx)) oligomerization without pharmacologic disruption, we have examined the transport and assembly of connexin constructs containing C-terminal di-lysine-based endoplasmic reticulum (ER) (HKKSL) or ER-Golgi intermediate compartment (AKKFF) targeting sequences. By immunofluorescence microscopy, Cx43-HKKSL transiently transfected into HeLa cells showed a predominantly ER localization, although Cx43-AKKFF was localized to the perinuclear region of the cell. Sucrose gradient analysis of Triton X-100-solubilized connexins showed that either Cx43-HKKSL or Cx43-AKKFF expressed alone by HeLa cells was maintained as an apparent monomer. In contrast to Cx43-HKKSL, Cx32-HKKSL was maintained in the ER as stable hexamers, consistent with the notion that Cx32 and Cx43 oligomerization occur in distinct intracellular compartments. Furthermore, Cx43-HKKSL and Cx43-AKKFF inhibited trafficking of Cx43 and Cx46 to the plasma membrane. The inhibitory effect was because of the formation of mixed oligomers between Cx43-HKKSL or Cx43-AKKF and wild type Cx43 or Cx46. Taken together, these results suggest that Cx43-HKKSL and Cx43-AKKFF recirculate through compartments where oligomerization occurs and may be maintained as apparent monomers by a putative Cx43-specific quality control mechanism.