RESUMO
Recombinant adenovirus vector systems have been used extensively in protein research and gene therapy. However, the construction and characterization of recombinant adenovirus is a tedious and time-consuming process. TIGIT is a recently discovered immunosuppressive molecule that plays an important role in maintaining immunological balance. The construction of recombinant adenovirus mediating TIGIT expression must be simplified to facilitate its use in the study of TIGIT. In this study, the TIGIT gene was combined with green fluorescent protein (GFP); the TIGIT-GFP gene was inserted into a gateway plasmid to construct a TIGIT-GFP adenovirus. HEK 293A cells were infected with the adenovirus, which was then purified and subjected to virus titering. TIGIT-GFP adenovirus was characterized by flow cytometry and immunofluorescence, and its expression in mouse liver was detected by infection through caudal vein injection. The results showed the successful construction of the TIGIT-GFP adenovirus (5 x 10(10) PFU/mL). Co-expression of TIGIT and GFP was identified in 293A and liver cells; synthesis and positioning of TIGIT-GFP was viewed under a fluorescence microscope. TIGIT-GFP was highly expressed on liver cells 1 day (25.53%) after infection and faded 3 days (11.36%) after injection. In conclusion, the fusion of TIGIT with GFP allows easy, rapid, and uncomplicated detection of TIGIT translation. The construction of a TIGIT-GFP adenovirus, mediating TIGIT expression in vitro and in vivo, lays the foundation for further research into TIGIT function and gene therapy. Moreover, the TIGIT-GFP adenovirus is a helpful tool for studying other proteins (which could replace the TIGIT gene).
Assuntos
Adenoviridae/genética , Vetores Genéticos/genética , Proteínas de Fluorescência Verde/genética , Receptores Imunológicos/genética , Proteínas Recombinantes de Fusão/genética , Animais , Linhagem Celular , Expressão Gênica , Ordem dos Genes , Vetores Genéticos/isolamento & purificação , Hepatócitos/metabolismo , Humanos , Camundongos , Transdução GenéticaRESUMO
OBJECTIVE: To explore the role of heat shock protein 20 (HSP20)-mediated cardiomyocyte exosomes in the cardiac function in mice with myocardial infarction via the activation of the protein kinase B (Akt) signaling pathway. MATERIALS AND METHODS: A total of 30 mice were enrolled to establish the model of myocardial infarction. Next, these mice were divided into three groups, namely Blank group (healthy mice), Model group (mouse models of myocardial infarction), and HSP20 group (mouse models of myocardial infarction transfected with lentivirus to overexpress HSP20). After that, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining assay was performed to detect myocardial apoptosis. Reactive oxygen species (ROS) accumulation in myocardial tissues was determined via dihydroethidium (DHE) staining assay. Western blotting was employed to analyze the expression level of Akt. The expression levels of inflammatory factors tumor necrosis factor-alpha (TNF-α) and interleukin 1 beta (IL-1ß) in HSP20-mediated cardiomyocyte exosomes were measured through quantitative real time polymerase chain reaction (qRT-PCR). RESULTS: Compared with that in Blank group, the number of cardiomyocyte exosomes was increased in Model group and HSP20 group under anoxic conditions (p<0.05). The results of quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) proved that the HSP20 messenger ribonucleic acid (mRNA) expression in mediated cardiomyocyte exosomes was significantly lower in Model group than that in Blank group (p<0.05), while in HSP20 group, it was overtly higher than that in Model group but clearly lowered compared with that in Blank group (p<0.05). The protein expression of Akt in cardiomyocyte exosomes was evidently decreased in Model group compared with that in Blank group (p<0.05), while it was notably increased in HSP20 group compared with that in Model group (p<0.05). In comparison with Blank group, Model group had significantly elevated mRNA expression levels of TNF-α and IL-1ß. The mRNA expression levels of TNF-α and IL-1ß in HSP20 group were remarkably lower than those in Model group (p<0.05). The results of TUNEL assay revealed that the overexpression of HSP20 affected myocardial apoptosis. The myocardial apoptosis index in Model group [(38.42±2.52) %] was higher than that in Blank group [(9.74±1.21) %], HSP20 group had a significantly decreased myocardial apoptosis index [(22.36±2.13) %] in comparison with Model group (p<0.05). In accordance with DHE staining comparison, the accumulation of ROS in myocardial tissues in Model group was significantly higher than that in Blank group (p<0.05) and HSP20 group (p<0.05). CONCLUSIONS: We demonstrated that HSP20-mediated cardiomyocyte exosomes activate the AKT signaling pathway, repress TNF-α and IL-1ß factors, and alleviate myocardial infarction.
Assuntos
Exossomos/metabolismo , Proteínas de Choque Térmico HSP20/metabolismo , Infarto do Miocárdio/patologia , Miócitos Cardíacos/ultraestrutura , Proteínas Proto-Oncogênicas c-akt/metabolismo , Animais , Apoptose , Células Cultivadas , Modelos Animais de Doenças , Exossomos/ultraestrutura , Proteínas de Choque Térmico HSP20/genética , Humanos , Interleucina-1beta/metabolismo , Masculino , Camundongos , Microscopia Eletrônica de Transmissão , Miocárdio/citologia , Miocárdio/patologia , Miócitos Cardíacos/metabolismo , Cultura Primária de Células , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais , Fator de Necrose Tumoral alfa/metabolismoRESUMO
OBJECTIVE: To investigate clinical effects of rosuvastatin on blood lipid levels, hemorheological profiles, vascular endothelial function, pentraxin 3 (PTX-3) level, the number of granule membrane glycoprotein (GMP-140) molecules and platelet aggregation rate in elderly patients with acute myocardial infarction (AMI) undergoing elective percutaneous coronary intervention (PCI). PATIENTS AND METHODS: Total of 120 elderly patients admitted with AMI undergoing elective PCI from July 2014 to January 2016 were selected. The patients were divided into the control group and the experimental group based on the rule of random number generation and double-blind controlled trial, 60 cases in each group. All of 120 patients were treated with routine medications; the experimental group was orally administered with rosuvastatin 1 week before PCI. Blood lipid levels, hemorheological profiles, vascular endothelial function, PTX-3, the number of GMP-140 molecules and platelet aggregation rate were compared between two groups before treatment with rosuvastatin and 10d after elective PCI. RESULTS: Triglycerides, plasma total cholesterol, and low-density lipoprotein levels were significantly lower (p<0.05) in the experimental group when compared with the control group; plasma viscosity, fibrinogen, the viscosity of blood in the high shear rates and in the low shear rates in the experimental group were significantly lower than those of the control group (p<0.05); FMD and NMD in the experimental group were significantly higher than those of the control group (p<0.05); ET-1, TXA2 levels in the experimental group were lower, however, PGI2, NO as well as NOS in the experimental group were higher, when compared the control group, the differences were statistically significant (p<0.05); PTX-3, the number of GMP-140 molecules and platelet aggregation rate in the experimental group were significantly lower than those of the control group (p<0.05). CONCLUSIONS: Oral administration of rosuvastatin 1 week before PCI can significantly improve the blood lipid levels and hemorheological profiles, enhance endothelial function, reduce the PTX-3 level and the number of GMP-140 molecules, decrease the platelet aggregation rate, therefore improving prognosis in elderly patients with AMI undergoing PCI.
Assuntos
Anticolesterolemiantes/farmacologia , Proteína C-Reativa/análise , Procedimentos Cirúrgicos Eletivos , Infarto do Miocárdio/terapia , Intervenção Coronária Percutânea , Agregação Plaquetária/efeitos dos fármacos , Rosuvastatina Cálcica/farmacologia , Componente Amiloide P Sérico/análise , Idoso , Método Duplo-Cego , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Infarto do Miocárdio/sangueRESUMO
A nonlinear first principle model is developed for a laboratory-scaled multivariable chemical reactor rig in this paper and the on-line model predictive control (MPC) is implemented to the rig. The reactor has three variables-temperature, pH, and dissolved oxygen with nonlinear dynamics-and is therefore used as a pilot system for the biochemical industry. A nonlinear discrete-time model is derived for each of the three output variables and their model parameters are estimated from the real data using an adaptive optimization method. The developed model is used in a nonlinear MPC scheme. An accurate multistep-ahead prediction is obtained for MPC, where the extended Kalman filter is used to estimate system unknown states. The on-line control is implemented and a satisfactory tracking performance is achieved. The MPC is compared with three decentralized PID controllers and the advantage of the nonlinear MPC over the PID is clearly shown.
Assuntos
Algoritmos , Reatores Biológicos , Indústria Química/instrumentação , Desenho de Equipamento/métodos , Análise de Falha de Equipamento/métodos , Retroalimentação , Modelos Teóricos , Indústria Química/métodos , Simulação por Computador , Sistemas Computacionais , Sistemas On-Line , Teoria de SistemasRESUMO
Human hair keratins were among the first to be studied but it is only recently that sufficient information has been obtained to gain a basic biologic perspective of these proteins. Hair keratins are members of the intermediate filament family of proteins, yet are sufficiently divergent from epidermal keratins to warrant separate classification: type Ia and IIa ("hard"/hair keratins) and type Ib and IIb (epidermal and other "soft" keratins). As with hair keratins from other species, the human proteins may be distinguished from their epidermal counterparts by a relatively higher cysteine content, 7.6% versus 2.9%, respectively. This feature reflects utilization of disulfide bonding in producing a tougher, more durable structure in the tissues in which the hair keratins are distributed. Although prominent in hair, their distribution is not strictly limited to this tissue. A number of molecular characteristics have been elucidated from human hair keratin gene studies including amino acid sequence data for a type Ia hair keratin. Studies of various pedigrees has revealed a fairly wide latitude of variation in human hair keratin expression that is tolerated without associated obvious hair phenotypic change. Thus, a foundation of knowledge regarding these proteins has emerged and continues to evolve.
Assuntos
Cabelo/química , Queratinas/análise , Sequência de Aminoácidos , Aminoácidos/análise , Humanos , Dados de Sequência MolecularRESUMO
The hair follicle provides an excellent system in which to study growth and differentiation. Hair keratins are useful tissue-specific molecular markers for these events. By comparing a second mouse Type I hair keratin cDNA clone, MHKA-2, with our previously described MHKA-1, we have been able to contrast the nucleotide sequences and corresponding deduced amino acid sequences of the smallest (mHa4) and the largest (mHa1) major Type I hair keratins. Both nucleotide sequences and both deduced amino acid sequences share high identity but have distinct segments suitable for generation of specific molecular probes. Comparison of amino acid sequences adjacent to the central helical domains has demonstrated homologous subdomains, designated H1 and H2, in the Type I hair keratin nonhelical termini. Although there is only 56% amino acid identity in the carboxy-terminal nonhelical domains, a common sequence, T-------CGPC----R, has been identified in this domain, suggesting a possible common functional role for this portion of the molecule. In addition, it appears that mHa4 may differ in part from mHa1 by deletion of a segment between the H2 subdomain and the conserved sequence. Staining of mouse and human hair follicles with AmHa1, a monospecific polyclonal antibody for mHa1, and AE13, an antibody specific for all Type I hair keratins, suggests differential expression of individual Type I hair keratins in both species. This supports our hypothesis that distinct functional requirements are satisfied by the multiplicity of hair keratins.
Assuntos
Cabelo/metabolismo , Queratinas/genética , Sequência de Aminoácidos , Animais , Anticorpos/imunologia , Especificidade de Anticorpos , Sequência de Bases , Clonagem Molecular , DNA/genética , Queratinas/imunologia , Queratinas/metabolismo , Camundongos , Dados de Sequência Molecular , Distribuição TecidualRESUMO
Although it has been shown previously that an acidic (type I) "soft" keratin can interact with many basic (type II) "soft" keratins to form 10-nm intermediate filaments, it has been unclear whether "soft" keratins are compatible with the "hard" keratins typically found in hair and nail. To address this issue and to generate more structural information about hard keratins, we have isolated and sequenced a cDNA clone that encodes a mouse hair basic keratin (b4). Our sequence data revealed new information regarding the structural conservation of hard keratins as a group, being significantly different from soft keratins. Using expression vectors containing appropriate cDNA inserts, we studied the expression of this basic (b4) as well as an acidic (a1) mouse hair keratin in HeLa cells. The expression of these alien hair keratins in the transfected cells was surveyed using a panel of monoclonal and polyclonal antibodies. Our results indicated that the basic and acidic hair keratin readily incorporated into the existing endogenous soft keratin network of HeLa cells. Overproduction of hair keratin, however, occasionally led to the formation of cytoplasmic aggregates containing both hard and soft keratins. These data suggest that although small amounts of newly synthesized hair keratins can incorporate into the "scaffolding" of the preformed soft keratin filament network, possibly through dynamic subunit exchange, overproduction of hard keratins can lead to the partial collapse of the soft keratin network. These observations, along with the deduced amino acid sequence data, support and extend the concept that hard and soft keratins, although closely related, are divergent enough to justify their being divided into two separate subgroups.
Assuntos
Células HeLa/fisiologia , Queratinas/genética , Animais , Anticorpos/imunologia , Especificidade de Anticorpos , Sequência de Bases , Células Cultivadas , Clonagem Molecular , DNA/genética , DNA/isolamento & purificação , Cabelo/metabolismo , Células HeLa/metabolismo , Humanos , Filamentos Intermediários/metabolismo , Filamentos Intermediários/fisiologia , Queratinas/biossíntese , Queratinas/imunologia , Camundongos , Dados de Sequência Molecular , Transfecção/genéticaRESUMO
Hair follicle formation and maintenance involve intimate interactions between follicular epithelial cells and a group of specialized mesenchymal cells known as the dermal papilla. Using the random primer polymerase chain reaction, we have identified an approximately 1.4 kb osteopontin mRNA that is present in large quantities in cultured rat vibrissa dermal papilla cells but undetectable in cultured rat skin fibroblasts. In situ hybridization showed that the osteopontin gene is expressed in dermal papilla cells of pelage follicles during catagen but not in anagen or telogen. As an acidic glycosylated RGD-containing extracellular matrix protein, osteopontin can function both as a cell attachment protein and as a soluble cytokine playing roles in signaling, cell migration, tissue survival, anti-inflammation, and T-cell-mediated cellular immunity. Our results indicate that the comparison of the mRNA of cultured dermal papilla cells and fibroblasts can lead to the identification of not only anagen-specific genes (e.g., nexin 1), but also a catagen-specific gene. We have thus provided evidence that specific genes are turned on during catagen, which is therefore not simply a passive "degenerative" phase. The functional role of osteopontin in catagen is unclear but it may promote the formation of a tightly aggregated dermal papilla, and/or protect the dermal papilla cells from apoptosis induced by cytokines or hypoxia during catagen.
Assuntos
Derme/citologia , Folículo Piloso/citologia , Folículo Piloso/crescimento & desenvolvimento , Sialoglicoproteínas/genética , Alopecia/fisiopatologia , Animais , Sequência de Bases , Células Cultivadas , Fibroblastos/citologia , Fibroblastos/fisiologia , Expressão Gênica/fisiologia , Mesoderma/citologia , Dados de Sequência Molecular , Osteopontina , RNA Mensageiro/análise , Ratos , Ratos Wistar , Vibrissas/citologiaRESUMO
Proteinases and their inhibitors are very likely to function as mediators or regulators of the hair growth cycle. Very little information is currently available, however, regarding the specific inhibitors present in human hair follicles at defined stages of their growth cycle. In this study we have analyzed two proteinase inhibitors, plasminogen activator inhibitor type 2 and protease nexin 1, in human hair follicles using in situ hybridization and/or immunohistochemistry. Protease nexin 1 mRNA was found only in the mesenchymal population of the hair follicle, i.e., the follicular papilla cells, during the anagen but not the catagen phase. In contrast, plasminogen activator inhibitor type 2 was localized to several epithelial populations in the follicle: the more differentiated cells of the infundibulum; the companion layer in anagen follicles; and the single layer of outer root sheath cells directly abutting the club hair in telogen follicles. At least some of the plasminogen activator inhibitor type 2 in human follicles appears to be in the relaxed form, as evidenced by strong staining with an antibody that is specific for this form of the inhibitor. This suggests that plasminogen activator inhibitor type 2 interacts with and is cleaved by an endogenous follicular proteinase and supports a constitutive role for this inhibitor in human follicular epithelia.
Assuntos
Proteínas de Transporte/análise , Folículo Piloso/química , Inibidor 2 de Ativador de Plasminogênio/análise , Serpinas/análise , Precursor de Proteína beta-Amiloide , Apoptose , Proteínas de Transporte/fisiologia , Humanos , Imuno-Histoquímica , Hibridização In Situ , Inibidor 2 de Ativador de Plasminogênio/genética , Inibidor 2 de Ativador de Plasminogênio/fisiologia , Nexinas de Proteases , RNA Mensageiro/análise , Receptores de Superfície Celular , Serpina E2RESUMO
When applied at the individual patch level, the classic competition-colonization models of species coexistence assume that propagules of superior competitors can displace adults of inferior competitors (displacement competition). But if adults are invulnerable to displacement by propagules (as trees are to seeds), and propagules compete to replace adults that die for reasons independent of the outcome of juvenile competition (a lottery system), a competition-colonization trade-off alone is not able to produce coexistence. However, we show that coexistence is possible if patch density varies spatially, such that it becomes a niche axis. We also show how a dispersal-fecundity trade-off can partition variation in patch density. We discuss the application of these models to empirical systems. An important implication of communities coexisting via variation in patch density is that the amount of habitat loss necessarily interacts with the pattern of loss in affecting extinctions, invasions, and coexistence, in contrast to displacement competition models, for which the spatial pattern of loss is not important or is less important. Finally, with respect to mechanisms promoting coexistence, we suggest that trade-offs between different stages of colonization could be far more common in nature than a trade-off between competitive ability and colonization ability.
RESUMO
Three iodine-substituted derivatives of cocaine, methyl esters of 3-[(2'-, 3'-, and 4'-iodobenzoyl)oxy]-8-methyl-[1R-(exo,exo)]-8- azabicyclo[3.2.1]octane-2 carboxylic acid (2a-c), were synthesized and subjected to N-demethylation to give the corresponding noriodococaines 3-[(2'-,3'-, and 4'-iodobenzoyl)oxy]-[1R-(exo,exo)]-8- azabicyclo[3.2.1]-octane-2-carboxylic acid (3a-c). These were remethylated with [11C]CH3I to give the [N-11C-methyl]iodococaines 4a-c which were examined in baboon brain in vivo using positron emission tomography (PET). Compared to [N-11C]cocaine itself the regional distributions were changed from a highly specific localization in the corpus striatum to more diffuse patterns which included the cerebellum and cortex. Peak brain uptakes and clearance kinetics were also changed. [N-11C]-o-Iodococaine (4a) had a peak uptake in the striatum at 4-5 min after injection of only 17% that of cocaine in the same animal. The peak uptake of [N-11C]-p-iodococaine (4c) was 60% of that of [N-11C]cocaine and a clearance half-time of approximately 55 min, twice that of [N-11C]cocaine. [N-11C]-m-Iodococaine (4b) displayed half the uptake of [N-11C]cocaine, buts its clearance was similar to that of the parent molecule. The fractions of unmetabolized tracer in blood plasma at 1-30 min were higher for 4a-c than for [N-11C]cocaine. Plasma protein binding experiments showed 10%, 0.3%, 1.6%, and 6% as the free fraction for cocaine and o-, m-, and p-iodococaines respectively, consistent with the low brain uptake observed for the ortho isomer, and implicated alpha 1-acid glycoprotein as responsible for the low free fraction of o-iodococaine. The potencies of 2a-c to displace tritiated cocaine from striatal membranes were p-iodo approximately cocaine greater than m-iodo approximately o-iodo.
Assuntos
Encéfalo/metabolismo , Radioisótopos de Carbono , Cocaína/análogos & derivados , Iodo , Animais , Ligação Competitiva , Proteínas Sanguíneas/metabolismo , Cerebelo/metabolismo , Cocaína/química , Cocaína/farmacocinética , Corpo Estriado/metabolismo , Feminino , Lobo Frontal/metabolismo , Marcação por Isótopo , Cinética , Papio , Ligação Proteica , Ratos , Relação Estrutura-Atividade , Distribuição Tecidual , Tomografia Computadorizada de EmissãoRESUMO
Fig wasps (Agaonidae: Hymenoptera) are seed predators and their interactions with Ficus species (Moraceae) range from mutualism to parasitism. Recently considerable attention has been paid to conflicts of interest between the mutualists and how they are resolved in monoecious fig species. However, despite the fact that different conflicts can arise, little is known about the factors that influence the persistence of the mutualism in functionally dioecious Ficus. We studied the fig pollinator mutualism in 14 functionally dioecious fig species and one monoecious species from tropical lowland rainforests near Madang, Papua New Guinea. Observations and experiments suggest that (i) pollinating wasps are monophagous and attracted to a particular host species; (ii) pollinating and non-pollinating wasps are equally attracted to gall (male) figs and seed (female) figs in functionally dioecious species; (iii) differing style lengths between gall figs and seed figs may explain why pollinators do not develop in the latter; (iv) negative density dependence may stabilize the interaction between pollinating wasps and their parasitoids; and (v) seed figs may reduce the search efficiency of non-pollinators. This increased pollinator production without a corresponding decrease in seed production could provide an advantage for dioecy in conditions where pollinators are limiting.
Assuntos
Comportamento Alimentar , Frutas/fisiologia , Frutas/parasitologia , Pólen/fisiologia , Rosales/fisiologia , Rosales/parasitologia , Vespas/fisiologia , Animais , Reprodução , SementesRESUMO
Many genetic defects are known to cause abnormal development of the coat in mice. Hair keratin genes would seem to be particularly promising candidates among the potential targets of these mutations in mice and of inherited hair-related abnormalities in humans as well. We used specific probes from cloned and sequenced mouse hair keratin cDNAs (MHKA-2, MHKB-1, and MHKB-2) to assess linkage of hair keratin genes and mouse mutations. We analyzed DNA from the progeny of interspecies backcrossed mice for segregation of hair mutations, hair ("hard") keratin alleles, and epidermal ("soft") keratin alleles (Krt-1 and Krt-2 loci). The results suggest that most, if not all, hair keratin genes (types Ia and IIa) are part of the Krt-1 locus on chromosome 11 and Krt-2 locus on chromosome 15, respectively. Linkage of the hair keratin genes and the mutations Re, Den, and Bsk on chromosome 11, and Ca, Sha, and Ve on chromosome 15 suggests that these mutations may possibly involve altered hair keratin expression or structure. In addition, the nondispersion of homologous keratin genes in the mammalian genome suggests that a domain organization of the genes has influenced evolution of the keratin gene family and that the organization may play a significant role in tissue-specific and developmental regulation of keratin gene expression as well.
Assuntos
Mapeamento Cromossômico , Cabelo/fisiologia , Queratinas/genética , Família Multigênica , Sequência de Aminoácidos , Animais , Sequência de Bases , Southern Blotting , DNA/genética , DNA/isolamento & purificação , Sondas de DNA , Epiderme/fisiologia , Biblioteca Gênica , Ligação Genética , Camundongos , Camundongos Mutantes , Dados de Sequência Molecular , Homologia de Sequência do Ácido NucleicoRESUMO
We isolated 1, Y1, Y2 chromosomes of the male M.m vaginalis by microdissection and amplified the DNA copies by DOP-PCR. Using the DOP-PCR products of 1, Y1, Y2 chromosomes of M.m vaginalis as template respectively and the primers designed within human SRY HMG box, the Sry gene of the male M.m vaginalis was amplified, and it was cloned and sequenced. It is proved that Y2 is the real sex chromosome in the male M.m vaginalis at molecular level for the first time, and Sry was localized on Y2 chromosome.
Assuntos
Mapeamento Cromossômico , Proteínas de Ligação a DNA/genética , Cervo Muntjac/genética , Proteínas Nucleares , Fatores de Transcrição , Cromossomo Y , Animais , Clonagem Molecular , Dissecação , Masculino , Reação em Cadeia da Polimerase , Proteína da Região Y Determinante do SexoRESUMO
The character of microscopic damage on the space structure of the herring sperm DNA by wetting treatment of acetic acid with Raman spectroscopic technique was reported. The Raman characteristic frequency and intensity were changed in different degree after wetting treatment on DNA by acetic acid. The results showed that the entire DNA molecule was damaged by acetic acid including backbone phosphodiester groups, deoxyribose and base stacking. The conformation and forms of DNA were changed, hydrogen bond was broken, B-form was modified, both single and double chains were disrupted. Base pairs were damaged. The damage degrees of the base pairs in turn were cytosine, thymine, guanine and adenine.
Assuntos
Ácido Acético/efeitos adversos , Dano ao DNA/efeitos dos fármacos , Animais , Quimioembolização Terapêutica , Peixes , Humanos , Neoplasias Hepáticas/terapia , Conformação de Ácido Nucleico , Sêmen/química , Análise Espectral RamanRESUMO
A group of specialized mesenchymal cells located at the root of the mammalian hair follicle, known as the follicular or dermal papillary cells, are involved in regulating the hair cycle, during which keratinocytes of the lower follicle undergo proliferation, degeneration and regrowth. Using the arbitrarily primed-PCR approach, we have identified a 1.3 kb messenger RNA that is present in large quantities in cultured rat follicular papillary cells, but not in skin fibroblasts. This mRNA encodes nexin 1, a potent protease inhibitor that can inactivate several growth-modulating serine proteases including thrombin, urokinase and tissue plasminogen activator. In situ hybridization showed that nexin 1 message is accumulated in the follicular papilla cells of anagen follicles, but is undetectable in keratinocytes or other skin mesenchymal cells. In addition, nexin 1 message level varies widely among several immortalized rat vibrissa papillary cell lines, and these levels correlate well with the reported abilities of these cell lines to support in vivo follicular reconstitution. These results suggest a possible role of nexin 1 in regulating hair follicular growth.
Assuntos
Proteínas de Transporte/genética , Folículo Piloso/enzimologia , Periodicidade , RNA Mensageiro/metabolismo , Inibidores de Serina Proteinase/genética , Precursor de Proteína beta-Amiloide , Animais , Sequência de Bases , Células Cultivadas , Feminino , Folículo Piloso/ultraestrutura , Dados de Sequência Molecular , Nexinas de Proteases , Ratos , Ratos Wistar , Receptores de Superfície CelularRESUMO
A fundamental question in ecology is how many species occur within a given area. Despite the complexity and diversity of different ecosystems, there exists a surprisingly simple, approximate answer: the number of species is proportional to the size of the area raised to some exponent. The exponent often turns out to be roughly 1/4. This power law can be derived from assumptions about the relative abundances of species or from notions of self-similarity. Here we analyze the largest existing data set of location-mapped species: over one million, individually identified trees from five tropical forests on three continents. Although the power law is a reasonable, zeroth-order approximation of our data, we find consistent deviations from it on all spatial scales. Furthermore, tropical forests are not self-similar at areas =50 hectares. We develop an extended model of the species-area relationship, which enables us to predict large-scale species diversity from small-scale data samples more accurately than any other available method.
RESUMO
Extensive neuroanatomical, neurophysiological, and behavioral evidence demonstrates that GABAergic neurons inhibit endogenous dopamine release in the mammalian corpus striatum. Positron emission tomography (PET) studies in adult female baboons, using the dopamine D2-specific radiotracer 11C-raclopride, were undertaken to assess the utility of this imaging technique for measuring these dynamic interactions in vivo. 11C-raclopride binding was imaged prior to and following the administration of either gamma-vinyl-GABA (GVG), a specific suicide inhibitor of the GABA-catabolizing enzyme GABA transaminase, or lorazepam, a clinically prescribed benzodiazepine agonist. Striatal 11C-raclopride binding increased following both GVG and lorazepam administration. This increase exceeded the test/retest variability of 11C-raclopride binding observed in the same animals. These findings confirm that changes in endogenous dopamine concentrations resulting from drug-induced potentiation of GABAergic transmission can be measured with PET and 11C-raclopride. Finally, this new strategy for noninvasively evaluating the functional integrity of neurophysiologically linked transmitter systems with PET supports its use as an approach for assessing the multiple mechanisms of drug action and their consequences in the human brain.
Assuntos
Encéfalo/metabolismo , Dopamina/metabolismo , Salicilamidas/metabolismo , Tomografia Computadorizada de Emissão , Ácido gama-Aminobutírico/farmacologia , Aminocaproatos/farmacologia , Animais , Radioisótopos de Carbono , Cerebelo/metabolismo , Corpo Estriado/metabolismo , Feminino , Lorazepam/farmacologia , Neurotransmissores/metabolismo , Papio , Racloprida , VigabatrinaRESUMO
The psychostimulant drug of abuse, cocaine (benzoylecgonine methyl ester), is rapidly metabolized by cleavage of its two ester groups, to give benzoylecgonine (BE) and ecgonine methyl ester, and by N-demethylation, to give N-norcocaine (NC). The recent use of [N-methyl-11CH3]cocaine to image brain cocaine binding sites with positron emission tomography (PET) raises the question of whether PET images partially reflect the distribution and kinetics of labeled cocaine metabolites. We prepared [O-methyl-11CH3]cocaine by methylation of the sodium salt of BE with [11C]CH3I, and showed that PET baboon brain scans, as well as regional brain kinetics and plasma time-activity curves corrected for the presence of labeled metabolites, are nearly identical to those seen with [N-methyl-11CH3]cocaine. This strongly suggests that 11C metabolites do not significantly affect PET images, because the metabolite pattern is different for the two labeled forms of cocaine. In particular, nearly half the 11C in blood plasma at 30 min was [11C]CO2 when [N-methyl-11CH3]cocaine was administered, whereas [11C]CO2 was not formed from [O-methyl-11CH3]cocaine. Only a trace of [11C]NC was detected in plasma after [O-methyl-11CH3]cocaine administration. Nearly identical brain PET data were also obtained when 4'-[N-methyl-11CH3]fluorococaine and 4'-[18F]fluorococaine (prepared by nucleophilic aromatic substitution from [18F]fluoride- and 4'-nitrococaine) were compared with [N-methyl-11CH3]cocaine. In vitro assays with rat brain membranes showed that cocaine and 4'-fluorococaine were equipotent at the dopamine reuptake site, but that 4'-fluorococaine was about 100 times more potent at the 5-hydroxytryptamine reuptake site.(ABSTRACT TRUNCATED AT 250 WORDS)