RESUMO
Ocular prostheses are liable to host microbial adhesion, which can favor conjunctival inflammation. Knowing the microbiota of the ocular prosthesis and anophthalmic socket is important for predicting infection-related pathogens. This study evaluated the microbiota of the prostheses and anophthalmic sockets of six individuals by 16S rRNA sequencing. The microbial abundance and diversity were analyzed using the Operational Taxonomic Units at the genus level. Forty-seven phyla, capturing 1,258 named genera, were recorded in the sample set. In both sites, the most frequent genera were Fusobacterium, Staphylococcus, Prevotella, and Streptococcus. The microbial abundance was higher for the anophthalmic socket than for the prosthesis. Alpha diversity showed no significant differences in bacterial richness or diversity between sites. Although the microbial abundance in the anophthalmic socket was higher, both sites had similar microbiota with high diversity and low dominance among the genera.
Assuntos
Olho Artificial , Microbiota , Humanos , RNA Ribossômico 16S/genética , Genes de RNAr , Biofilmes , Microbiota/genéticaRESUMO
OBJECTIVE: Mesiodentes are usually found in the central position of the upper or lower jaw as supernumerary teeth. Here, we obtained 10 mesiodentes and three permanent teeth (PT) and separated the dental pulp (DP) from these into crown and root portions. We then characterized and compared the isolated crown portion-derived cells (crown cells) with root portion-derived cells (root cells) using a range of in vitro assays. MATERIALS AND METHODS: Crown cells and root cells were examined for cell surface marker expression, colony-forming unit-fibroblast (CFU-F), cell proliferation, cell cycle characteristics and markers, and osteogenic and adipogenic differentiation. RESULTS: The proportion of CD105-positive cells (CD105(+) cells) in the crown cells vs the root cells varied among the mesiodentes, but not among the PT. When there were more CD105(+) cells in the root cells than in the crown cells, the root cells showed higher CFU-F, proliferation capacity, and osteogenic differentiation capacity. In contrast, when the crown cells contained more CD105(+) cells than the root cells, the crown cells showed the higher CFU-F, proliferation capacity, and osteogenic differentiation capacity. In addition, the sorted CD105(+) cells showed higher CFU-F and proliferation capacity than the sorted CD105(-) cells. CONCLUSION: These results indicated that proportion of CD105(+) cells is an effective means of characterizing DP-derived cells in mesiodentes.
Assuntos
Polpa Dentária/citologia , Células-Tronco Mesenquimais/citologia , Coroa do Dente/citologia , Raiz Dentária/citologia , Dente Supranumerário/patologia , Adolescente , Antígenos de Superfície/imunologia , Criança , Pré-Escolar , Ensaio de Unidades Formadoras de Colônias , Feminino , Citometria de Fluxo , Humanos , Masculino , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Adulto JovemRESUMO
AIM: To evaluate the antibiofilm activity of sodium hypochlorite (NaOCl) and chlorhexidine (CHX) solutions associated with cetrimide (CTR), and QMiX using confocal laser scanning microscopy. METHODOLOGY: Enterococcus faecalis (ATCC- 29212) biofilms were induced on bovine dentine blocks for 14 days. The dentine blocks containing biofilm were immersed for 1 min in the following solutions: 2.5% NaOCl; 2.5% NaOCl + 0.2% CTR; 2% CHX; 2% CHX + 0.2% CTR; 0.2% CTR; QMiX. After contact with the solutions, the dentine blocks were stained with Live/Dead(®) BacLight for analysis of the remaining biofilm using confocal laser scanning microscope. Images were evaluated using the BioImage_L software to determine the total biovolume (µm(3) ), the green biovolume (live cells) (µm(3) ) and the percentage of substrate coverage (%). The data were subjected to nonparametric statistical test using Kruskal-Wallis and Dunn's tests at 5% significance level. RESULTS: After exposure to irrigants, the total biovolume observed for CHX, CHX+CTR, CTR, QMiX was similar to distilled water (P > 0.05). NaOCl and NaOCl+CTR had the lowest total and green biovolume. The CTR and QMiX had intermediate green biovolume, with greater antibacterial activity than CHX and CHX+CTR (P < 0.05). The NaOCl and NaOCl+CTR solutions were associated with microorganism removal and substrate cleaning ability. CONCLUSIONS: NaOCl and NaOCl+CTR solutions were effective on microorganism viability and were able to eliminate biofilm. The addition of cetrimide did not influence antibacterial activity.
Assuntos
Biofilmes , Compostos de Cetrimônio/química , Irrigação Terapêutica , Animais , Bovinos , Cetrimônio , Microscopia ConfocalRESUMO
PURPOSE: Photodynamic therapy (PDT) is effective in reducing pathogenic microorganisms in the oral cavity and in preventing dental diseases. This study evaluated the pre-irradiation time using PDT (diode laser associated with 0.01% methylene blue) to decrease the number of microorganisms in the visible plaque in permanent teeth. METHODS: This randomized clinical trial included 108 homologous lower permanent first molars (36 and 46) with biofilm from 54 children aged six to 12 years. PDT was performed (0.01% methylene blue photosensitizer/therapeutic laser-InGaAIP), according to the following protocols: Group 1, biofilm collection of the distal area of the lingual surface of 36 µm before PDT; group 2, mesial area of the lingual surface of 36 µm 1 min after PDT; group 3, area of the lingual surface of 46 µm before PDT; and group 4, mesial area of the lingual surface of 46 µm 5 min after PDT. RESULTS: After statistical analysis, significant differences were observed between the groups (p = 0.000). In groups 2 and 4, the number of bacteria tended to decrease, with a more evident bacterial reduction in group 4. CONCLUSIONS: Pre-irradiation reduced the number of colony-forming units of mature bacterial biofilms in vivo. A time of 5 min resulted in a greater reduction in the number of colony-forming units. CLINICAL TRIAL REGISTRATION: ReBEC Identifier: RBR-6bqfp3; Date of Register: March 2nd, 2020. Retrospectively Registered.
Assuntos
Anti-Infecciosos , Fotoquimioterapia , Criança , Humanos , Azul de Metileno/uso terapêutico , Fotoquimioterapia/métodos , Biofilmes/efeitos da radiação , Fármacos Fotossensibilizantes/uso terapêuticoRESUMO
BACKGROUNDS: Maternal employment has been shown to be associated with childhood overweight and obesity (Ow/Ob), but the presence of family members who care for children in place of the mothers might influence children's Ow/Ob and lifestyles. The influence of maternal employment on children's Ow/Ob should be examined together with the presence of caregivers such as grandparents. OBJECTIVES: The effects of maternal employment and the presence of grandparents on lifestyles and Ow/Ob in Japanese pre-school children were investigated. DESIGN/SUBJECTS: Cross-sectional study on 2114 children aged 3-6 years who attended all childcare facilities in a city and primary caregivers was conducted. MEASUREMENTS: Children's weight and height, family environments (family members, maternal employment, single parent, number of siblings and parental Ow/Ob) and lifestyles (dietary, physical activity and sleeping habits) were surveyed using a self-administered questionnaire. Ow/Ob was defined by the International Obesity Task Force cut-offs. RESULTS: The eligible participants were 1765 children. The prevalence of Ow/Ob was 8.4% in boys and 9.9% in girls. Maternal employment was associated positively with irregular mealtimes, unfixed snacking times, bedtime after 10 p.m. and nighttime sleep duration of less than 10 h, whereas three-generation families were associated negatively with irregular mealtimes after adjustment for children's characteristics and family environments. Irregular mealtimes (OR (95% CI); 2.03 (1.36, 3.06)) and nighttime sleep duration of less than 10 h (1.96 (1.28, 3.01)) were associated with increased risks of being Ow/Ob. Both maternal employment and three-generation families were significantly associated with children's Ow/Ob. However, three-generation families maintained a significant association (1.59 (1.08, 2.35)) after adjustment for maternal employment. CONCLUSIONS: These study results suggest that the grandparents who care for pre-school children in place of mothers are more likely to contribute to childhood Ow/Ob than maternal employment. The family-focused lifestyle strategies to prevent childhood Ow/Ob must include grandparents who care for children.
Assuntos
Educação Infantil , Características da Família , Mães/estatística & dados numéricos , Obesidade/epidemiologia , Mulheres Trabalhadoras/estatística & dados numéricos , Criança , Comportamento Infantil , Educação Infantil/tendências , Pré-Escolar , Estudos Transversais , Dieta/tendências , Emprego , Feminino , Humanos , Japão/epidemiologia , Estilo de Vida , Masculino , Obesidade/etiologia , Sobrepeso/epidemiologia , Linhagem , Prevalência , Fatores SocioeconômicosRESUMO
In this study, the fouling potentials of polysaccharides contained in mixed liquor suspension in a membrane bioreactor (MBR) treating municipal wastewater were investigated using lectin affinity chromatography. This investigation was carried out with different membranes to assess the effects of membrane materials on fouling potentials of polysaccharides. In lectin affinity chromatography, some polysaccharides with high affinity to the lectin in the column can be retained in the column. The fouling potentials of the retained polysaccharides were evaluated by dead-end filtration test. Degree of reduction in fouling potential differed considerably when different lectins were used in affinity chromatography indicating that fouling potentials of polysaccharide differed depending on types of polysaccharide. Trends in the reduction of fouling potential were different depending on membrane materials. Characteristics of the organic matter associated with polysaccharides removed by lectins were investigated by means of excitation-emission matrices (EEM). The results of EEM analysis indicate that the characteristics of the organic matter eluted from different lectins were different as long as elution reagents for the lectin were different. Characteristics of the organic matter eluted from the lectins which have the same elution reagent were similar in terms of shapes of EEM fluorescence spectra. However, the trends in reduction of fouling potentials could not be explained by the characteristics of organic matter assessed by EEM analysis. On the basis of the results obtained in this study, it can be concluded that characteristics of membrane and structures or properties of sugar chain would play an important role in determining fouling potentials.
Assuntos
Reatores Biológicos , Cromatografia de Afinidade/métodos , Membranas Artificiais , Lectinas de Plantas/química , Polissacarídeos/química , Eliminação de Resíduos Líquidos/métodos , Projetos PilotoRESUMO
A high-dispersion spectrum of Comet C/1999S4 (LINEAR) was obtained in the optical region with the high-dispersion spectrograph on the Subaru telescope when the comet was 0.863 astronomical units from the Sun before its disintegration. We obtained high signal-to-noise ratio emission lines of the cometary NH2 bands from which an ortho-to-para ratio (OPR) of 3.33 +/- 0.07 was derived on the basis of a fluorescence excitation model. Assuming that cometary NH2 mainly originates from ammonia through photodissociation, the derived OPR of NH2 molecules should reflect that of ammonia, which provides information on the environment of molecular formation or condensation and of the thermal history of cometary ices. Assuming that the OPR of ammonia in comets was unchanged in the nucleus, the derived spin temperature of ammonia (28 +/- 2 kelvin) suggests that a formation region of the cometary ammonia ice was between the orbit of Saturn and that of Uranus in the solar nebula.
Assuntos
Amônia , Meteoroides , Gelo , Análise Espectral , TemperaturaRESUMO
BACKGROUND: Inadequately reprocessed reusable surgical instruments (RSIs) may harbour infectious agents which may then be transferred to a suitable site for replication. AIM: To determine the cumulative effect of 20 cycles of contamination, cleaning (manual or manual followed by automated) and steam sterilization on high-complex-design RSIs used for orthopaedic surgery. METHODS: New flexible medullary reamers and depth gauges were contaminated by soaking in tryptone soya broth, containing 5% sheep blood and 109 cfu/mL of Staphylococcus aureus (ATCC 25923), for 5 min. To mimic a worse-case scenario, RSIs were dried 7 h and subjected to either (a) rinsing in distilled water, (b) manual cleaning or (c) manual plus automated cleaning (reference standard), and steam sterilization. The contamination, cleaning, and sterilization cycle was repeated 20 times. Adenosine triphosphate (ATP) was measured after cleaning procedures; microbial load and residual protein were measured following the 10th and 20th reprocessing, in triplicate. Scanning electron microscopy (SEM) was used to confirm soil and biofilm presence on the RSIs after the 20th reprocessing. FINDINGS: Manual and manual plus automated cleaning significantly reduced the amount of ATP and protein residues for all RSIs. Viable bacteria were not detected following sterilization. However, SEM detected soil after automated cleaning, and soil, including biofilms, after manual cleaning. CONCLUSION: Soil and/or biofilms were evident on complex-design RSIs following 20 cycles of contamination and reprocessing, even using the reference standard method of cleaning. Although the depth gauges could be disassembled, biological residues and biofilm accumulated in its lumen. The current design of these RSIs prevents removal of all biological soil and this may have an adverse effect on patient outcome.
Assuntos
Carga Bacteriana , Descontaminação/métodos , Procedimentos Ortopédicos/instrumentação , Staphylococcus aureus/isolamento & purificação , Esterilização/métodos , Instrumentos Cirúrgicos/microbiologia , Trifosfato de Adenosina/análise , Humanos , Microscopia Eletrônica de VarreduraRESUMO
Hydrographic data collected from research cruises, bottom-anchored moorings, drifting Ice-Tethered Profilers, and satellite altimetry in the Beaufort Gyre region of the Arctic Ocean document an increase of more than 6,400 km3 of liquid freshwater content from 2003 to 2018: a 40% growth relative to the climatology of the 1970s. This fresh water accumulation is shown to result from persistent anticyclonic atmospheric wind forcing (1997-2018) accompanied by sea ice melt, a wind-forced redirection of Mackenzie River discharge from predominantly eastward to westward flow, and a contribution of low salinity waters of Pacific Ocean origin via Bering Strait. Despite significant uncertainties in the different observations, this study has demonstrated the synergistic value of having multiple diverse datasets to obtain a more comprehensive understanding of Beaufort Gyre freshwater content variability. For example, Beaufort Gyre Observational System (BGOS) surveys clearly show the interannual increase in freshwater content, but without satellite or Ice-Tethered Profiler measurements, it is not possible to resolve the seasonal cycle of freshwater content, which in fact is larger than the year-to-year variability, or the more subtle interannual variations.
RESUMO
In recent clinical studies, contamination of the inner parts of dental implants through bacterial penetration along the implant components has been observed. The aim of the present in-vitro study was to investigate leakage of Fusobacterium nucleatum through the interface between implants and premachined or cast abutments. Both premachined (n=10) and cast (n=10) implant-abutment assemblies were inoculated with 3.0 microL of microbial inoculum. The assemblies were completely immersed in 5.0 mL of tryptic soy broth culture medium to observe leakage at the implant-abutment interface after 14 days of anaerobic incubation. Bacterial growth in the medium, indicative of microbial leakage, was found only in 1 out of 9 samples (11.1%) in each group. Both premachined and cast abutments connected to external hexagonal implants provide low percentages of bacterial leakage through the interface in in vitro unloaded conditions if the manufacturer's instructions and casting procedures are properly followed.
Assuntos
Dente Suporte/microbiologia , Implantes Dentários/microbiologia , Infiltração Dentária/microbiologia , Planejamento de Prótese Dentária , Fusobacterium nucleatum/crescimento & desenvolvimento , Anaerobiose , Ligas de Cromo/química , Meios de Cultura , Ligas Dentárias/química , Técnica de Fundição Odontológica , Humanos , Teste de Materiais , Propriedades de Superfície , Fatores de TempoRESUMO
The PCDD/DFs and coplanar PCBs (co-PCBs) in sediment samples from Gwangyang Bay in South Korea was investigated. The total concentration of dioxins and their toxic equivalent quantity (TEQ; calculated with the WHO 2005 Toxic Equivalency Factors) value in the surface sediment of the outer site (261 pg g(-1) TOC, 4.4 pg-TEQ g(-1)) were 3-fold higher than the inner site (90 pg g(-1) TOC, 1.1 pg-TEQ g(-1)) in the Bay. The dioxin in the sediment samples was found to come from a mixture of the impurities of pentachlorophenol (PCP), chloronitrofen (CNP) and combustion based on the result of hierarchical cluster analysis (HCA). These dioxin sources have been influenced by the characterization associated with this region which was both an agricultural-centered and industrial-centered area. According to principal component analysis (PCA) related to the Kow values for the congener-specific composition of co-PCBs in the sediment core, the Kanechlor (KC)-500 and the atmospheric deposition were identified as the possible sources. The maximum burden in the sediment core was 1.3 kg for 1967-1974 and the total burdens of PCDD/DFs and co-PCBs in the sediment core were estimated to be 6.6 kg during the past 50 years. The cumulative burdens of dioxin are still increasing in Gwangyang Bay.
Assuntos
Benzofuranos/análise , Sedimentos Geológicos/análise , Bifenilos Policlorados/análise , Dibenzodioxinas Policloradas/análogos & derivados , Poluentes Químicos da Água/análise , Dibenzofuranos Policlorados , Monitoramento Ambiental , Coreia (Geográfico) , Dibenzodioxinas Policloradas/análiseRESUMO
OBJECTIVE: To evaluate by means of Petrifilmtrade mark system (3M, St Paul, MN, USA) the level of bacterial contamination in water from old and new dental units (air-water syringes and high-speed turbines) before and after flushing water through waterlines. The old dental units had been used for 13 years and the new dental units for 1 year. A fast method named Petrifilmtrade mark AC (3M) was employed to evaluate the level of water contamination with total aerobic bacteria and Petrifilmtrade mark EC for Escherichia coli and coliforms. METHODS: Dental unit water were collected before and after flushing of 4 (air-water syringes) and 2 min (high-speed turbines) from 24 old and new dental units. Thereafter, samples were diluted, inoculated onto Petrifilmtrade mark plates and incubated. RESULTS: The filtered tap water that filled up dental unit reservoirs showed a low level of bacterial contamination (4 and 15 CFU ml(-1)). However, all water samples from old and new dental units were highly contaminated. The flushing of dental unit waterlines reduced the bacterial count in all dental unit water, but the reduction was better in water from new dental units than from old dental units. E. coli and coliforms were not detected in any water samples analysed. CONCLUSION: Flushing water is a simple measure that should do part of dental routine, because it was able to reduce the level of total aerobic bacteria in water from old and new dental units.
Assuntos
Contagem de Colônia Microbiana/instrumentação , Descontaminação/métodos , Equipamentos Odontológicos/microbiologia , Microbiologia da Água , Purificação da Água/métodos , Bactérias Aeróbias , Brasil , Contaminação de Equipamentos , Reutilização de Equipamento , Estatísticas não ParamétricasRESUMO
Estrogenic activity was determined in sediments collected from Tokyo Bay. Sampling was performed at five stations including the site near the sewage treatment plant. The most estrogenic sediment collected near the sewage treatment plant was fractionated into ten fractions using normal-phase high-performance liquid chromatography. Chemical analysis was carried out for each fraction and nonylphenol (NP, 20,700ngg(-1)drywt) was detected at a higher concentration than estron (2.39ngg(-1)drywt) and 17beta-estradiol (<0.7ng g(-1)dry wt). Furthermore, each fraction was administered to male mummichogs (Fundulus heteroclitus), and vitellogenin (Vtg) was measured after two weeks. The induction of Vtg was observed; this estrogenic potency could be attributed to the NP content in this fraction. This is the first report to suggest that the high NP concentration in the sediments from Tokyo Bay has the potential to induce Vtg in wild fish.
Assuntos
Estrogênios/toxicidade , Fundulidae/sangue , Sedimentos Geológicos , Vitelogeninas/sangue , Poluentes Químicos da Água/toxicidade , Animais , Monitoramento Ambiental , Estradiol/análise , Estradiol/toxicidade , Estrogênios/análise , Estrona/análise , Estrona/toxicidade , Sedimentos Geológicos/análise , Japão , Masculino , Fenóis/análise , Fenóis/toxicidade , Poluentes Químicos da Água/análiseRESUMO
Multifractal analysis of cardiovascular variability series is an effective tool for the characterization of pathological states associated with congestive heart failure (CHF). Consequently, variations of heartbeat scaling properties have been associated with the dynamical balancing of nonlinear sympathetic/vagal activity. Nevertheless, whether vagal dynamics has multifractal properties yet alone is currently unknown. In this study, we answer this question by conducting multifractal analysis through wavelet leader-based multiscale representations of instantaneous series of vagal activity as estimated from inhomogeneous point process models. Experimental tests were performed on data gathered from 57 CHF patients, aiming to investigate the automatic recognition accuracy in predicting survivor and non-survivor patients after a 4 years follow up. Results clearly indicate that, on both CHF groups, the instantaneous vagal activity displays power-law scaling for a large range of scales, from ≃ 0.5s to ≃ 100s. Using standard SVM algorithms, this information also allows for a prediction of mortality at a single-subject level with an accuracy of 72.72%.
Assuntos
Insuficiência Cardíaca , Algoritmos , Frequência Cardíaca , Humanos , Sobreviventes , Nervo VagoRESUMO
Na(v)2/NaG is a putative sodium channel, whose physiological role has long been an enigma. We generated Na(v)2 gene-deficient mice by inserting the lacZ gene. Analysis of the targeted mice allowed us to identify Na(v)2-producing cells by examining the lacZ expression. Besides in the lung, heart, dorsal root ganglia, and Schwann cells in the peripheral nervous system, Na(v)2 was expressed in neurons and ependymal cells in restricted areas of the CNS, particularly in the circumventricular organs, which are involved in body-fluid homeostasis. Under water-depleted conditions, c-fos expression was markedly elevated in neurons in the subfornical organ and organum vasculosum laminae terminalis compared with wild-type animals, suggesting a hyperactive state in the Na(v)2-null mice. Moreover, the null mutants showed abnormal intakes of hypertonic saline under both water- and salt-depleted conditions. These findings suggest that the Na(v)2 channel plays an important role in the central sensing of body-fluid sodium level and regulation of salt intake behavior.
Assuntos
Comportamento Animal/fisiologia , Sistema Nervoso Central/metabolismo , Canais de Sódio/fisiologia , Cloreto de Sódio na Dieta/metabolismo , Administração Oral , Animais , Comportamento Animal/efeitos dos fármacos , Sistema Nervoso Central/embriologia , Diuréticos/farmacologia , Ingestão de Líquidos/fisiologia , Marcação de Genes , Genes Reporter , Heterozigoto , Homozigoto , Óperon Lac , Camundongos , Camundongos Knockout , Dados de Sequência Molecular , Especificidade de Órgãos , Proteínas Proto-Oncogênicas c-fos/metabolismo , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/genética , Solução Salina Hipertônica/administração & dosagem , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Canais de Sódio/genética , Sulfonamidas , Paladar/genética , Sede/fisiologia , Canais de Sódio Disparados por VoltagemRESUMO
Multiscale analysis of human heartbeat dynamics has been proved effective in characterizeing cardiovascular control physiology in health and disease. However, estimation of multiscale properties can be affected by the interpolation procedure used to preprocess the unevenly sampled R-R intervals derived from the ECG. To this extent, in this study we propose the estimation of wavelet coefficients and wavelet leaders on the output of inhomogeneous point process models of heartbeat dynamics. The RR interval series is modeled using probability density functions (pdfs) characterizing and predicting the time until the next heartbeat event occurs, as a linear function of the past history. Multiscale analysis is then applied to the pdfs' instantaneous first order moment. The proposed approach is tested on experimental data gathered from 57 congestive heart failure (CHF) patients by evaluating the recognition accuracy in predicting survivor and non-survivor patients, and by comparing performances from the informative point-process based interpolation and non-informative spline-based interpolation. Results demonstrate that multiscale analysis of point-process high-resolution representations achieves the highest prediction accuracy of 65.45%, proving our method as a promising tool to assess risk prediction in CHF patients.
Assuntos
Insuficiência Cardíaca/diagnóstico , Eletrocardiografia , Insuficiência Cardíaca/mortalidade , Insuficiência Cardíaca/fisiopatologia , Frequência Cardíaca/fisiologia , Humanos , Contração Miocárdica , Medição de Risco , Processamento de Sinais Assistido por Computador , Sobreviventes , Análise de OndaletasRESUMO
The sagittal split ramus osteotomy (SSRO) is a surgical technique used widely to treat many congenital and acquired mandibular discrepancies. Stabilization of the osteotomy site and the potential for skeletal relapse after the procedure are still major problems. The aim of this study was to compare the mechanical stability of six methods of rigid fixation in SSRO using a biomechanical test model. Sixty polyurethane replicas of human hemimandibles were divided into six groups. In group I, the osteotomies were fixed with two four-hole titanium miniplates; in group II, with one four-hole miniplate; in group III, with one four-hole miniplate+a bicortical screw; in group IV, with a grid miniplate; in group V, with a four-hole locking miniplate; and in group VI, with a six-hole miniplate. A linear load in the premolar region was applied to the hemimandibles. The resistance forces (N) needed to displace the distal segment by 1, 3, and 5mm were recorded and the data transmitted from the load cell to a computer. One-way analysis of variance with Tukey's post hoc test was performed to compare the means between groups. For the three displacement conditions, there was a strong tendency for the 2.0-mm plate+screw and the grid plate to have higher values.
Assuntos
Técnicas de Fixação da Arcada Osseodentária , Osteotomia Sagital do Ramo Mandibular/métodos , Fenômenos Biomecânicos , Placas Ósseas , Parafusos Ósseos , Humanos , Modelos Anatômicos , PoliuretanosRESUMO
A response of growth hormone (GH) to thyrotropin-releasing hormone (TRH) is observed in lower mammals and patients with diseases such as a cromegaly, but not in normal subjects. We have previously demonstrated the existence of intact TRH receptor mRNA in GH-secreting adenoma. To examine whether intact somatotrophs in the anterior pituitary also express TRH receptor, we attempted to localize both TRHR mRNA and GH immunoreactivity simultaneously. In situ hybridization analysis revealed TRHR mRNAs specifically in the anterior pituitary, and 61.1% of the anterior pituitary cells expressed this transcript. Staining for GH and PRL on the same samples showed that the somatotrophs apparently expressed TRHR mRNA and approximately 62.3% and 30.9% of hybridization-positive cells were somatotorophs and mammotrophs, respectively. Moreover, TRHR mRNA level in the somatotrophs expressed as the number of silver grains per cell was equivalent to that in the mammotrophs. These findings demonstrated expression of the TRHR mRNA in somatotrophs in the rat anterior pituitary, and also showed that more than 50% of the TRHR mRNA detected in the anterior pituitary was derived from these cells.
Assuntos
Expressão Gênica , Hormônio do Crescimento/metabolismo , Adeno-Hipófise/metabolismo , RNA Mensageiro/metabolismo , Receptores do Hormônio Liberador da Tireotropina/genética , Animais , Autorradiografia , Northern Blotting , Hormônio do Crescimento/análise , Hibridização In Situ , Masculino , Prolactina/análise , RNA Mensageiro/análise , Ratos , Ratos WistarRESUMO
The mammalian brain contains many species of proteoglycan. To identify each proteoglycan species, we have raised monoclonal antibodies against soluble chondroitin sulfate proteoglycans purified from 10-day-old rat brains. One monoclonal antibody, named monoclonal antibody 1G2, recognized two proteoglycan species with 220,000 and 150,000 mol. wt core glycoproteins (chondroitin sulfate proteoglycan-220 and chondroitin sulfate proteoglycan-150). Partial amino acid sequences of N-termini of their core proteins coincided with those of neurocan, a brain-unique chondroitin sulfate proteoglycan species, whose complete coding sequence was recently reported [Rauch et al. (1992) J. biol. Chem. 269, 19,536-19,547]. Western blots revealed that chondroitin sulfate proteoglycan-220 became detectable in the rat cerebrum on embryonic day 14, and that it disappeared from the brain around postnatal day 30. In contrast, a fairly large amount of chondroitin sulfate proteoglycan-150 remained in the mature brain. Immunohistochemical studies revealed that 1G2 antigen was first localized in the preplate zone, then both in the marginal zone and in the subplate of the rat cerebrum on embryonic day 16, prior to arrival of the first thalamic afferents at the cortex. On embryonic day 20, immunolabeling with monoclonal antibody 1G2 began to spread from the subplate into the developing cortical plate. On postnatal day 10, the neuropil of the cerebrum, except for the barrel field, was diffusely stained with the antibody, intensely in the hippocampus and superficial layers (I-III) of the cerebral cortex and weakly elsewhere. The barrel hollows were stained very weakly compared with the barrel walls at this stage. The immunoreactivity in the hippocampus and superficial cortical layers was weakened in the mature brain, so that no particular staining pattern, but weak and diffuse staining was observed in the adult rat cerebrum. The 1G2 antigen was immunohistochemically associated largely with glial fibrillary acidic protein-positive cells in primary cultures of the neonatal rat cerebrum. Both chondroitin sulfate proteoglycan-220 and chondroitin sulfate proteoglycan-150 were detected in the conditioned media not only of highly enriched cultures of fetal rat cortical neurons but also of pure cultures of mature astrocytes; more (12- to 20-fold) in the astrocyte conditioned media. Astrocytes, in addition to neurons, may be a cellular source of neurocan in brain at least under certain physiological conditions. The spaciotemporal expression pattern of 1G2 epitope-bearing proteoglycan, or neurocan, suggests that this proteoglycan species plays some roles at least in forming the elongation pathway for early cortical afferent fibers as well as the functional barrel structure in the somatosensory cortex.
Assuntos
Anticorpos Monoclonais , Córtex Cerebral/crescimento & desenvolvimento , Córtex Cerebral/metabolismo , Proteoglicanas de Sulfatos de Condroitina/biossíntese , Proteínas do Tecido Nervoso/biossíntese , Sequência de Aminoácidos , Animais , Astrócitos/metabolismo , Western Blotting , Células Cultivadas , Córtex Cerebral/citologia , Condroitina Liases/metabolismo , Proteoglicanas de Sulfatos de Condroitina/química , Proteoglicanas de Sulfatos de Condroitina/imunologia , Meios de Cultivo Condicionados , Eletroforese em Gel de Poliacrilamida , Feminino , Glucosidases/química , Hidrólise , Imuno-Histoquímica , Lectinas Tipo C , Camundongos , Camundongos Endogâmicos C3H/imunologia , Dados de Sequência Molecular , Proteínas do Tecido Nervoso/química , Proteínas do Tecido Nervoso/imunologia , Neurocam , Gravidez , Proteoglicanas/biossíntese , Proteoglicanas/química , Proteoglicanas/imunologia , Ratos , Ratos Sprague-DawleyRESUMO
A monoclonal antibody was obtained by immunizing mice with an extract of monkey brain. The monoclonal antibody 473 stained a small subpopulation of neurons in various regions of monkey and rat central nervous system. The perimeters of neuronal somata and the proximal parts of dendrites bound the antibody. Electron microscopic analysis showed that the immunoreactivity was associated with the outer surface of the cell. The immunoreactivity in the rat cerebral cortex appeared gradually during the second four weeks after birth. The antibody stained fetal cartilages but otherwise was specific to the nervous system. Experiments on the stability of the immunoreactivity to enzymatic and chemical treatments of the sections suggest that the antigen molecule is of proteoglycan nature.