ABSTRACT
Covalent organic frameworks (COFs) are promising electrocatalyst platforms owing to their designability, porosity, and stability. Recently, COFs with various chemical structures are developed as efficient electrochemical CO2 reduction catalysts. However, controlling the morphology of COF catalysts remains a challenge, which can limit their electrocatalytic performance. Especially, while porphyrin COFs show promising catalytic properties, their particle size is mostly large and uncontrolled because of the severe aggregation of crystallites. In this work, a new synthetic methodology for rationally downsized COF catalyst particles is reported, where a tritylated amine is employed as a novel protected precursor for COF synthesis. Trityl protection provides high solubility to a porphyrin precursor, while its deprotection proceeds in situ under typical COF synthesis conditions. Subsequent homogeneous nucleation and colloidal growth yield smaller COF particles than a conventional synthesis, owing to suppressed crystallite aggregation. The downsized COF particles exhibit superior catalytic performance in electrochemical CO2 reduction, with higher CO production rate and faradaic efficiency compared to conventional COF particles. The improved performance is attributed to the higher contact area with a conductive agent. This study reveals particle size as an important factor for the evaluation of COF electrocatalysts and provides a strategy to control it.
ABSTRACT
Although stereochemical control of carbon centers is a well-established technique in modern synthetic chemistry, that of tetrahedral metal complexes with a stereogenic metal center remains difficult due to the dynamic nature of their coordination bonds. Here we report the synthesis of a tetrahedral d8 high-spin chiral-at-nickel(II) complex composed exclusively of achiral ligands and the supramolecular control of its temperature-dependent spontaneous resolution in crystals. Under certain conditions, complex molecules with the same absolute configuration of the nickel(II) center grow into conglomerate crystals with a helically arranged structure due to intermolecular hydrogen bonding. This process is highly spontaneous, does not require any chiral sources, and is closely related to the origin of homochirality in biological systems. The obtained enantiopure nickel(II) complex will be a new type of redox-active chiral source for asymmetric synthetic chemistry.
ABSTRACT
Single-atom catalysts dispersed on support materials show excellent heterogeneous catalytic properties that can be tuned using the interactions between the single atoms and the support. Such interactions depend on whether the single atoms are located on the surface or within the interior of the support. However, little is known about immobilising single atoms on the surface or within the interior of supports deliberately and selectively. Herein, such location-selective placement of single atoms is achieved through the choice of metal complex precursor, solvent, and workup procedure. Using CdSe nanoplatelets as a support, a cis-[PtCl2(SO(CH3)2)2] precursor in an aprotic solvent exclusively attaches single Pt atoms on the surface of the support. In contrast, a [PtCl4]2- precursor in a protic solvent followed by amine treatment places 60% of the single Pt atoms inside the support by cation substitution. The surface-adsorbed single Pt atoms show higher stability in photocatalytic hydrogen evolution than the substituted ones, and the preclusion of substitution as internal Pt maximises the activity. Thus, this study provides a viable strategy for the structurally precise synthesis and design of single-atom catalysts.
Subject(s)
Amines , Nanoparticles , Ions , Catalysis , SolventsABSTRACT
Diquafosol tetrasodium (DQS), a purinergic P2Y2 receptor agonist, stimulates secretion of both water and mucins from the conjunctiva into tears. Hence, DQS-containing eye drops have been approved as a therapeutic option for dry eye disease in some Asian countries, including Japan. Recent clinical reports state that instilling DQS-containing eye drops significantly increases the lipid layer thickness in tears. Therefore, we examined this compound's direct actions on holocrine lipid-secreting meibomian gland cells and their function. Isolated meibomian gland cells (meibocytes) were procured from rabbits and cultivated in serum-free culture medium. Differentiated meibocytes with pioglitazone were used for the subsequent experiments. Intracellular Ca2+ signalling of the cells was dramatically elevated with DQS addition in a dose-dependent manner. This DQS-induced elevation was almost completely cancelled by the coexistence of the selective P2Y2 receptor antagonist AR-C118925XX. DQS treatment also facilitated total cholesterol (TC) release from cells into the medium. This effect of DQS on TC was suppressed significantly by the intracellular Ca2+ chelator BAPTA-AM as well as by AR-C118925XX. DNA fragmentation analysis revealed that DQS may have enhanced the apoptotic DNA fragmentation caused spontaneously by cells. Thus, DQS could stimulate meibocytes to release lipids through the P2Y2 receptor and possibly facilitate holocrine cell maturation.
Subject(s)
Cholesterol/metabolism , Meibomian Glands/metabolism , Ophthalmic Solutions/pharmacology , Polyphosphates/pharmacology , Receptors, Purinergic P2Y2/metabolism , Uracil Nucleotides/pharmacology , Animals , Cells, Cultured , Dry Eye Syndromes/pathology , Meibomian Glands/cytology , Purinergic P2Y Receptor Agonists/pharmacology , RNA, Messenger/genetics , Rabbits , Receptors, Purinergic P2Y2/genetics , Tears/chemistryABSTRACT
Chiral metal complexes show promise as asymmetric catalysts and optical materials. Chiral-at-metal complexes composed of achiral ligands have expanded the versatility and applicability of chiral metal complexes, especially for octahedral and half-sandwich complexes. However, Werner-type tetrahedral complexes with a stereogenic metal centre are rarely used as chiral-at-metal complexes because they are too labile to ensure the absolute configuration of the metal centre. Here we report the asymmetric construction of a tetrahedral chiral-at-zinc complex with high configurational stability, using an unsymmetric tridentate ligand. Coordination/substitution of a chiral auxiliary ligand on zinc followed by crystallisation yields an enantiopure chiral-only-at-zinc complex (> 99% ee). The enantiomer excess remains very high at 99% ee even after heating at 70 °C in benzene for one week. With this configurationally stable zinc complex of the tridentate ligand, the remaining one labile site on the zinc can be used for a highly selective asymmetric oxa-Diels-Alder reaction (98% yield, 87% ee) without substantial racemisation.
ABSTRACT
Despite efficient and specific in vitro knockdown, more reliable and convenient methods for in vivo knockdown of target genes remain to be developed particularly for retinal research. Using commercially available and chemically modified siRNA so-called Accell siRNA, we established a novel in vivo gene silencing approach in the rat retina. siRNA designed for knockdown of the house keeping gene Gapdh or four retinal cell type-specific genes (Nefl, Pvalb, Rho and Opn1sw) was injected into the vitreous body, and their retinal mRNA levels were quantified using real-time PCR. Intravitreal injection of siRNA for Gapdh resulted in approximately 40-70% reduction in its retinal mRNA levels, which lasted throughout a 9-day study period. Furthermore, all the selected retinal specific genes were efficiently down-regulated by 60-90% following intravitreal injection, suggesting injected siRNA penetrated into major retinal cell types. These findings were consistent with uniform distribution of a fluorescence-labeled siRNA injected into the vitreous body. Interestingly, gene silencing of Grin1, a core subunit of NMDA receptor, was accompanied by significant prevention from NMDA-induced retinal ganglion cell death. Thus, we provide single intravitreal injection of Accell siRNA as a versatile technique for robust and sustainable in vivo retinal gene silencing to characterize their biological functions under physiological and pathophysiological conditions.
Subject(s)
Gene Silencing/drug effects , Genetic Therapy , RNA, Small Interfering/pharmacology , Receptors, N-Methyl-D-Aspartate/genetics , Retinal Diseases/therapy , Animals , Genes, Essential/genetics , Humans , Intravitreal Injections , RNA, Double-Stranded/genetics , RNA, Double-Stranded/pharmacology , Rats , Receptors, N-Methyl-D-Aspartate/antagonists & inhibitors , Retina/pathology , Retinal Diseases/genetics , Retinal Diseases/pathology , Retinal Ganglion Cells/drug effects , Retinal Ganglion Cells/pathology , Vitreous Body/drug effectsABSTRACT
Self-assembled metal-organic architectures have great potential to undergo major structural changes into different architectures. Such molecular transformation is widely applicable to responsive systems like drug delivery and allosteric catalysis. A great number of metal-organic architectures responsive to a specific stimulus have been reported so far. However, interconversion between a pair of distinct metal-organic structures in response to multiple stimuli is rarely reported despite its high versatility. Herein we report multi-stimuli-responsive interconversion between a bowl-shaped and a capsule-shaped self-assembled ZnII complexes, [ZnII4L3X6] and [ZnII4L4], respectively, which were found to form in equilibrium from porphyrin-based ligand L and ZnII ions with different stoichiometry. Specifically, this interconversion was induced by four distinct external stimuli: exogenous ligands, Brønsted base/acid, solvents, and guest molecules. The mechanisms of the interconversion system are discussed in detail focusing on the species included in the equilibria. Thus, these findings would provide a helpful clue to design principles for multi-stimuli-responsive systems with functional versatility.
ABSTRACT
Hetero-multinuclear metal complexes are a promising class of compounds applicable to photoluminescence, magnetism, and catalysis. In this work, we have developed a synthetic method for hetero-tetranuclear metal complexes by combining advantages of site-selective redox switching and transmetalation. First, a homo-tetranuclear CoII4 complex was converted to a mixed-valence CoIIICoII3 complex by site-selective oxidation, which was then transmetalated from CoII to NiII to form a heterometallic CoIIINiII3 complex. Finally, a CoIINiII3 complex was synthesized by metal-selective reduction on the CoIII site. The basic structural frameworks of the main products in the whole process starting from the CoII4 complex are isostructural. Notably, the CoIINiII3 complex was not accessible by direct mixing of ligand, CoII, and NiII. This method would provide an alternative strategy for highly selective synthesis of hetero-multinuclear metal complexes.
ABSTRACT
Fibrodysplasia ossificans progressiva (FOP) is a rare autosomal dominant disorder characterized by congenital malformation of the great toes and by progressive heterotopic bone formation in muscle tissue. Recently, a mutation involving a single amino acid substitution in a bone morphogenetic protein (BMP) type I receptor, ALK2, was identified in patients with FOP. We report here that the identical mutation, R206H, was observed in 19 Japanese patients with sporadic FOP. This mutant receptor, ALK2(R206H), activates BMP signaling without ligand binding. Moreover, expression of Smad1 and Smad5 was up-regulated in response to muscular injury. ALK2(R206H) with Smad1 or Smad5 induced osteoblastic differentiation that could be inhibited by Smad7 or dorsomorphin. Taken together, these findings suggest that the heterotopic bone formation in FOP may be induced by a constitutively activated BMP receptor signaling through Smad1 or Smad5. Gene transfer of Smad7 or inhibition of type I receptors with dorsomorphin may represent strategies for blocking the activity induced by ALK2(R206H) in FOP.
Subject(s)
Activin Receptors, Type I/metabolism , Bone Morphogenetic Protein Receptors, Type I/metabolism , Cell Differentiation , Matrix Metalloproteinases, Secreted/metabolism , Myositis Ossificans/metabolism , Osteoblasts/metabolism , Osteogenesis , Signal Transduction , Smad1 Protein/metabolism , Smad5 Protein/metabolism , Activin Receptors, Type I/genetics , Amino Acid Substitution , Animals , Bone Morphogenetic Protein Receptors, Type I/genetics , Cell Line , Female , Humans , Male , Matrix Metalloproteinases, Secreted/genetics , Mice , Mutation, Missense , Myositis Ossificans/genetics , Myositis Ossificans/pathology , Osteoblasts/pathology , Pyrazoles/pharmacology , Pyrimidines/pharmacology , Smad1 Protein/genetics , Smad5 Protein/genetics , Smad7 Protein/genetics , Smad7 Protein/metabolismABSTRACT
Rats with neonatal ventral hippocampal lesions (NVHL) have been studied as a neurodevelopmental animal model of schizophrenia. NVHL rats exhibit postpubertal emergence of hyperresponsiveness to stress, suggesting increased mesolimbic dopamine (DA) activity. However, previous studies have not yielded clear evidence of this. Disturbances in the gamma-amino-butyric acid (GABA)-ergic system as well as the dopaminergic system are thought to be present in schizophrenia. To determine whether GABA(A) receptors play a role in the abnormal postpubertal behavior in NVHL rats, we compared changes in expression of mRNA of GABA(A) receptor subunits and in [(35)S] t-butylbicyclophosphorothionate ([(35)S] TBPS) binding in the prepubertal and postpubertal periods. Male pups were lesioned with ibotenic acid at postnatal day 7 (PD 7), and in situ hybridization and quantitative autoradiography were then performed. In NVHL rats, alpha1 subunit mRNA expression in prefrontal cortex was decreased at PD 35 (prepubertal period; by 21.7%), but increased at PD 56 (postpubertal period; by 21.4%) when compared with sham controls. beta2 subunit mRNA expression was increased in PFC in the postpubertal period (by 24.3%). beta3 subunit mRNA expression was increased in the caudate-putamen in the postpubertal period (by 37.2%). [(35)S] TBPS binding was increased in PFC only in the postpubertal period (by 17.7%). These findings suggest that dysfunction of the GABAergic system exists in NVHL rats. Furthermore, developmental and regional changes in GABA(A) receptor expression appear to occur in compensation for the attenuation of GABAergic system activity in NVHL rats.
Subject(s)
Corpus Striatum/metabolism , Hippocampus/physiopathology , Neuronal Plasticity/physiology , Prefrontal Cortex/metabolism , Receptors, GABA-A/genetics , gamma-Aminobutyric Acid/metabolism , Adaptation, Physiological/physiology , Aging/metabolism , Animals , Animals, Newborn , Binding, Competitive/physiology , Bridged Bicyclo Compounds, Heterocyclic/metabolism , Bridged Bicyclo Compounds, Heterocyclic/pharmacokinetics , Corpus Striatum/growth & development , Corpus Striatum/physiopathology , Denervation , Disease Models, Animal , Hippocampus/injuries , Male , Neural Pathways/growth & development , Neural Pathways/metabolism , Neural Pathways/physiopathology , Prefrontal Cortex/growth & development , Prefrontal Cortex/physiopathology , Protein Subunits/genetics , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , Receptors, GABA-A/metabolism , Schizophrenia/metabolism , Schizophrenia/physiopathology , Up-Regulation/physiologyABSTRACT
This study was undertaken to determine whether human oral keratinocyte stem cells characteristically express higher levels of the low-affinity neurotrophin receptor p75 and to elucidate the function of p75 in oral keratinocytes. Examination of their expression patterns and cell-cycling status in vivo showed that p75 was exclusively expressed in the basal cell layer of both the tips of the papillae and the deep rete ridges. These immunostaining patterns suggest a cluster organization; most p75(+) cells did not actively cycle in vivo. Cell sorting showed that cells in the p75(+) subset were smaller and possessed higher in vitro proliferative capacity and clonal growth potential than the p75(-) subset. Clonal analysis revealed that holoclone-type (stem cell compartment), meroclone-type (intermediate compartment), and paraclone-type (transient amplifying cell compartment) cells, previously identified in skin and the ocular surface, were present in human oral mucosal epithelium. Holoclone-type cells showed stronger p75 expression at both the mRNA and protein level than did meroclone- and paraclone-type cells. Among the several neurotrophins, nerve growth factor (NGF) and neurotrophin-3 stimulated p75(+) oral keratinocyte cell proliferation, and only NGF protected them from apoptosis. Our in vivo and in vitro findings indicate that p75 is a potential marker of oral keratinocyte stem/progenitor cells and that some neurotrophin/p75 signaling affects cell growth and survival.
Subject(s)
Keratinocytes/cytology , Receptor, Nerve Growth Factor/physiology , Stem Cells/cytology , Stem Cells/physiology , Apoptosis , Cell Culture Techniques , Cell Cycle , Cell Fractionation , Clone Cells , Colony-Forming Units Assay , Flow Cytometry , Gene Expression Regulation , Humans , Keratinocytes/physiology , Mouth Mucosa/cytology , Mouth Mucosa/physiology , Receptor, Nerve Growth Factor/deficiency , Receptor, Nerve Growth Factor/genetics , Reference Values , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Repeated administration of phencyclidine (PCP) induces behavioral sensitization to dopaminergic neural transmission. This phenomenon has been implicated in the pathophysiology of schizophrenia. Recently, GABAergic agonists have been shown to reduce behavioral activity induced by enhanced dopaminergic neural transmission, which is mediated by the GABA(A)/benzodiazepine (BZD) receptor complex. Thus, to investigate the role of BZD receptors during induction and expression of behavioral sensitization in PCP-sensitized animals, the effects of both single and repeated PCP administration on BZD receptors in rat brain were examined using in vitro quantitative autoradiography. Repeated PCP administration failed to significantly alter levels of [3H]flunitrazepam (FNZ) binding in any of the regions examined. However, significant increases in levels of [3H]FNZ binding were found in the nucleus accumbens and ventral pallidum 1 h after single administration of PCP. These results suggest that BZD binding sites may not play important roles in the development of PCP-induced sensitization at several sites of GABA(A)/BZD receptor complex, while changes in GABA function in the nucleus accumbens differ from other areas following single administration of N-methyl-D-aspartate (NMDA) antagonist.
Subject(s)
Brain/metabolism , Excitatory Amino Acid Antagonists/pharmacology , Flunitrazepam/metabolism , GABA Modulators/metabolism , Phencyclidine/pharmacology , Animals , Autoradiography , Behavior, Animal/drug effects , Brain/drug effects , Excitatory Amino Acid Antagonists/administration & dosage , Image Processing, Computer-Assisted , Limbic System/drug effects , Limbic System/metabolism , Male , Phencyclidine/administration & dosage , Rats , Rats, Wistar , Stereotyped Behavior/drug effectsABSTRACT
PURPOSE: Immunohistochemically, all cells in the porcine corneal epithelium, from the superficial to the basal layer, express integrin beta1. Flow cytometric study has shown that they comprise integrin beta1 high-expressing (beta1 2+) and low-expressing (beta1+) populations. This study was undertaken to determine their proliferation characteristics. METHODS: Epithelial cells from porcine corneas were sorted and labeled with anti-integrin beta1 antibody and a fluorescent-dye-conjugated secondary antibody. The fluorescent intensity of labeled cells was analyzed and beta1 2+ and beta1+ cells were cultured in an adhesive-coated culture plate. RESULTS: Flow cytometry demonstrated that the epithelial cells comprised two distinct populations with a similar ratio throughout the cornea. Whereas beta1 2+ cells attached and grew to confluence in the plate, beta1+ attached only transiently to the plate and exhibited minimal growth. CONCLUSIONS: The data indicate that the porcine cornea contains two distinct populations of epithelial cells, one exhibiting high and the other low integrin beta1 expression. The observation that beta1 2+ cells had greater growth potential suggests that they may represent an enriched population of transit-amplifying cells.
Subject(s)
Epithelium, Corneal/cytology , Epithelium, Corneal/metabolism , Integrin beta1/metabolism , Animals , Cell Culture Techniques , Cell Division , Cell Separation , Flow Cytometry , Fluorescent Antibody Technique, Indirect , Stem Cells , SwineABSTRACT
Human beta-defensins (hBDs), which are mainly expressed in epithelial tissues, may contribute to infection-protective mechanisms in the ocular surface. We examined hBD1 and hBD2 expression in the ocular surface by RT-PCR and immunohistochemistry and studied the effects of immunosuppressive agents on their expression in human corneal epithelial cells in vitro. mRNA expression of hBD1 and hBD2 was confirmed in corneal epithelial cells. While the hBD1 peptide was immunohistochemically detected in corneal, limbal, and conjunctival epithelium, the level of expression was stronger in limbal- and conjunctival- than in corneal epithelium. Very weak expression of the hBD2 peptide was detected only in corneal epithelium. After 48-h culture of human corneal epithelial cells in the presence of 10(-5) M dexamethasone or 1 mg l(-1) cyclosporin A, total RNA was extracted and hBD1 and hBD2 mRNA expressions compared using semi-quantitative RT-PCR and introduced amplified fragment length polymorphism (iAFLP) assay. The two methods yielded almost identical results. hBD1 mRNA expression was not changed by dexamethasone but was down-regulated by cyclosporin A. hBD2 mRNA expression was up-regulated by dexamethasone and down-regulated by cyclosporin A. Our findings suggest that hBD1 and hBD2 are regulated by different mechanisms and that hBD1 may contribute to infection-protective mechanisms in the relatively immunosuppressed status induced by dexamethasone.
Subject(s)
Cyclosporine/pharmacology , Dexamethasone/pharmacology , Epithelium, Corneal/drug effects , Immunosuppressive Agents/pharmacology , beta-Defensins/metabolism , Adult , Cells, Cultured , Epithelium, Corneal/metabolism , Gene Expression Regulation/drug effects , Humans , Male , Polymorphism, Genetic , RNA, Messenger/genetics , Reverse Transcriptase Polymerase Chain Reaction , beta-Defensins/geneticsABSTRACT
PURPOSE: To reexamine whether the alpha5 chain of type IV (alpha5[IV]) collagen, thought to be absent, is in fact present in human amniotic membrane. METHODS: Cryosections of human amniotic membrane obtained at Cesarean section were immunohistochemically examined for the presence of alpha5(IV), with or without inclusion of the denaturing step. Amniotic membrane was digested with collagenase to release the noncollagenous NC1 domain from the alpha-chain. The NC1 domain of alpha5(IV) was then assayed on Western blot analysis. Identical experiments were performed with human corneas and conjunctivae obtained from an American eye bank. RESULTS: The basement membrane of denatured samples of amniotic membrane and cornea stained positive for alpha5(IV). Without the denaturing step, only corneal samples were positive. With or without denaturing, conjunctival epithelium did not stain. Western blot analysis detected NC1 domains of alpha5(IV) in amniotic membrane and corneal samples. CONCLUSIONS: The basement membrane of amniotic membrane resembles that of corneal epithelium but not conjunctiva. Amniotic membrane may be an excellent substrate for corneal epithelial cells.
Subject(s)
Amnion/metabolism , Collagen Type IV/metabolism , Epithelium, Corneal/metabolism , Basement Membrane/metabolism , Blotting, Western , Conjunctiva/metabolism , Female , Fluorescent Antibody Technique, Indirect , HumansABSTRACT
Rabbit immunoglobulin G (IgG) binds specifically to the human corneal epithelium. We investigated this phenomenon to identify the binding material in the tissue. Sections of human cornea were immunostained with negative-control grade rabbit IgG, F(ab')(2) (the antigen recognition fragment of IgG yielded by pepsin digestion), and Fc fragments. Human corneal epithelium was homogenized in a buffer containing Triton X-100 and the water-insoluble residue was subjected to two-dimensional gel electrophoresis. Western blotting was performed on the gels to detect materials that bound with rabbit IgG Fc fragments. Spots in the gel that bound with the Fc fragments were collected and analysed by peptide mass fingerprinting (PMF). Using an antibody against a binding candidate, keratin 5 (suggested by PMF analysis), we checked the occurrence of this phenomenon. Immunohistochemistry showed that rabbit IgG bound to corneal epithelium via the Fc region but not F(ab')(2). Western blots using rabbit Fc fragments showed that some protein spots in the gel bound to the fragments. All of the binding materials were basic and had molecular weights of approximately 55 kDa. PMF analysis showed that the binding material was keratin 5; all binding materials on the transferred membrane were recognized by an anti-keratin 5 antibody. On corneal sections, anti-keratin 5 produced staining similar to that of rabbit IgG. These findings suggest that rabbit IgG binds to keratin 5 in the human corneal epithelium via the Fc region of the molecule.
Subject(s)
Epithelium, Corneal/immunology , Immunoglobulin Fc Fragments/metabolism , Immunoglobulin G/metabolism , Keratins/metabolism , Animals , Electrophoresis, Gel, Two-Dimensional , Epithelium, Corneal/metabolism , Humans , In Vitro Techniques , Keratin-5 , Peptide Mapping , Protein Binding , RabbitsABSTRACT
PURPOSE: To determine the feasibility of using human amniotic membrane (AM) as a substrate for culturing oral epithelial cells and to investigate the possibility of using autologous cultivated oral epithelial cells in ocular surface reconstruction. METHODS: An ocular surface injury was created in one eye of each of eight adult albino rabbits by a lamellar keratectomy, and a conjunctival excision was performed, including and extending 5 mm outside the limbus. Oral mucosal biopsy specimens were obtained from these eight adult albino rabbits and cultivated for 3 weeks on a denuded AM carrier. The cultivated epithelium was examined by electron microscopy (EM) and immunohistochemically labeled for several keratins. At 3 to 4 weeks after the ocular surface injury, the conjunctivalized corneal surfaces of the eight rabbits were surgically reconstructed by transplanting the autologous cultivated oral epithelial cells on the AM carrier. RESULTS: The cultivated oral epithelial sheet had four to five layers of stratified, well-differentiated cells. EM revealed that the epithelial cells were very similar in appearance to those of normal corneal epithelium, had numerous desmosomal junctions, and were attached to a basement membrane with hemidesmosomes. Immunohistochemistry confirmed the presence of the keratin pair 4 and 13 and keratin-3 in the cultivated oral epithelial cells. Corneas that were grafted with the cultivated oral epithelial cells on an AM carrier were clear and were all epithelialized 10 days after surgery. CONCLUSIONS: Cultures of oral epithelial cells can be generated to confluence on AM expanded ex vivo from biopsy-derived oral mucosal tissue. Autologous transplantation was performed with these cultivated oral epithelial cells onto the ocular surfaces of keratectomized rabbit eyes. Autologous transplantation of cultivated oral epithelium is a feasible method for ocular surface reconstruction. The long-term outcome of such transplantation is not yet clear, and its feasibility in clinical use should be evaluated further.
Subject(s)
Amnion , Cell Culture Techniques/methods , Corneal Injuries , Epithelial Cells/transplantation , Eye Injuries/surgery , Mouth Mucosa/cytology , Animals , Cell Division , Disease Models, Animal , Epithelial Cells/metabolism , Epithelial Cells/ultrastructure , Feasibility Studies , Fluorescent Antibody Technique, Indirect , Humans , Keratins/metabolism , Rabbits , Transplantation, AutologousABSTRACT
To make clear the distinctive odorants of fish sauce, addition tests were carried out with respect to deodorized fish sauce samples. Volatiles from the headspace gas of nontreated fish sauce and deodorized fish sauce were characterized by gas chromatography-olfactometry and gas chromatography-mass spectrometry. Four compounds contributing to the distinctive odor of fish sauce were 2-methylpropanal, 2-methylbutanal, 2-ethylpyridine, and dimethyl trisulfide. Addition tests (quantitative descriptive analysis) with respect to the four odorants showed that contributors to the fishy aroma were 2-ethylpyridine and dimethyl trisulfide and that all four odorants contributed to the sweaty aroma. Furthermore, the fecal note of fish sauce was essential with 2-ethylpyridine and dimethyl trisulfide, and the rancid note was essential with all four odorants.