RESUMO
Virtual reality (VR)-based training has shown some benefits in medical education, supporting skill acquisition, and helping reduce anxiety in real-world settings. However, the use of VR simulators in veterinary education remains limited. This study aimed to introduce a VR simulator to support veterinarian training in canine anaesthesia induction and endotracheal intubation. This study involved a group that learned solely with instructional videos (video group), and one that learned concurrently with the video and VR simulator (VR group). Third- and fourth-year veterinary students were included and underwent a descriptive test on canine endotracheal intubation. Canine endotracheal intubation success rates were compared between the video (n = 364) and VR (n = 60) groups of fifth-year students. A survey on the VR usability was conducted (n=91). The median descriptive test scores improved in the VR (63.3/100) vs the video group (51.5/100). The canine intubation success rates were comparable in the VR and video groups at 84.3â¯% and 77.4â¯%, respectively. A total of 90.1â¯% of the surveyed students rated the ease of use of the simulator highly. Overall, VR simulators were well-received, suggesting benefits in new skill retention. Further studies are required to evaluate the extent of skill improvement through VR-based training, compared to conventional methods, and to assess its impact on student motivation. Evaluating the long-term effects of VR-based training on skill development and retention will also provide a deeper understanding of its educational benefits.
RESUMO
Age-related macular degeneration (AMD) is a vision-threatening disease characterized by choroidal fibrovascular membrane (FVM) formation, choroidal neovascularization (CNV) and choroidal fibrosis. No safe and effective therapeutic method has been developed for the choroidal fibrosis, although anti-vascular endothelial growth factor therapy can partially shrink the CNV. We recently reported that periostin (POSTN), which is produced by retinal pigment epithelial cells, has an important role in the formation of preretinal FVMs, but its role in choroidal FVMs has not been determined. In this study, we used Postn knockout mice to investigate the role played by POSTN in choroidal FVM formation. In addition, we used a new class of RNA interference (RNAi) agent (NK0144) that targets POSTN and determined its effect on choroidal FVM development. Genetic ablation of Postn had an inhibitory effect not only on CNV formation but also on choroidal fibrosis in a mouse CNV model. NK0144 also had a greater inhibitory effect on both the CNV and choroidal fibrosis than control RNAi with no apparent adverse effects. These findings suggest a causal relationship between POSTN and choroidal FVM formation, and also a potential therapeutic role of intravitreal NK0144 for AMD.
Assuntos
Moléculas de Adesão Celular/genética , Neovascularização de Coroide/terapia , Degeneração Macular/terapia , Interferência de RNA , Animais , Sequência de Bases , Adesão Celular , Moléculas de Adesão Celular/metabolismo , Movimento Celular , Proliferação de Células , Células Cultivadas , Corioide/irrigação sanguínea , Corioide/patologia , Técnicas de Silenciamento de Genes , Terapia Genética , Humanos , Injeções Intravítreas , Camundongos Endogâmicos C57BL , Camundongos Knockout , Dados de Sequência Molecular , RNA Interferente Pequeno/administração & dosagem , RNA Interferente Pequeno/genética , Epitélio Pigmentado da Retina/patologia , Epitélio Pigmentado da Retina/fisiologia , Receptor 3 Toll-Like/metabolismoRESUMO
Here we described our strategies to attain a better prognosis for hepatocellular carcinoma (HCC) patients by surgery. Among a variety of attempts conducted to date, we focused on anatomical resection and intraoperative contrast-enhanced ultrasonography. There are still controversies with respect to the significance of anatomical resection. We analyzed the significance of this surgical procedure in 207 patients without macrovascular invasion. These patients underwent either anatomical resection or non-anatomical resection. We found that the patients with anatomical resection had higher recurrence-free survival rate than those with non-anatomical resection. Univariable analysis showed that liver damage, the serum level of alpha-fetoprotein, tumor number, surgical margin, and type of surgery (anatomical or non-anatomical resection) were significant predictive factors for intrahepatic recurrence. Multivariable analysis revealed that multiple tumors, alpha-fetoprotein, and non-anatomical resection were independent risk factors for recurrence. We conclude that anatomical resection is a recommendable surgical procedure in patients without macrovascular invasion. A recent innovation is the development of contrast-enhanced ultrasonography. Then we have applied this to liver surgery intraoperatively. We confirm that vascular images contribute to a precise diagnosis and the detection of small portal tumor thrombi, and that Kupffer images are useful to discover the minute tumors. In addition, by clarifying the relationship between tumors and the vascular architecture, real-time 3-dimensional images using Kupffer imaging are a promising guide during the surgical procedures, although further development is needed.
Assuntos
Carcinoma Hepatocelular/cirurgia , Hepatectomia , Neoplasias Hepáticas/cirurgia , Ultrassonografia de Intervenção , Idoso , Carcinoma Hepatocelular/diagnóstico por imagem , Carcinoma Hepatocelular/patologia , Feminino , Humanos , Neoplasias Hepáticas/diagnóstico por imagem , Neoplasias Hepáticas/patologia , Masculino , Recidiva Local de Neoplasia/diagnóstico , Prognóstico , Taxa de SobrevidaRESUMO
BACKGROUND: Although a wait of several seconds after clamping is recommended when an automatic stapler is used to achieve adequate hemostasis, this wait has not been experimentally clarified. METHODS: To determine whether waiting is necessary between clamping and firing of a linear stapler, this study evaluated the number of staple line bleeding points and histologic changes in stapling sites of porcine small intestine (n = 46). It also assessed the ratio of dry to wet tissue weight (DW ratio) (n = 20) of porcine small intestine clamped between the prongs of a linear stapler. The sites were studied separately as follows: no wait with a four-row device (n = 12), no wait with a six-row device (n = 11), wait with a four-row device (n = 12), and wait with a six-row device (n = 11). The linear stapler was fired immediately after clamping in the no wait group and 1 min after clamping in the wait group. RESULTS: The mean number of staple line bleeding points in 2 to 5 min with the six-row device and in 3 to 5 min with the four-row device after firing were significantly less in the wait group than in the no wait group using the same device (p < 0.05). Cross sections of staple lines showed a higher frequency of mucosal cutting in the no wait group than in the wait group for both the four-row and the six-row devices (both significant at p < 0.01). Although the mean wet tissue weights of anastomotic sites did not change in either group, the mean DW ratio was significantly less in the wait group than in the no wait group (p < 0.01). CONCLUSIONS: A 1-min interval after clamping decreases the amount of clamped tissue. Waiting may thus be necessary to reduce bleeding from stapling sites, which may be related to a decrease in mucosal cutting.
Assuntos
Intestino Delgado/patologia , Intestino Delgado/cirurgia , Laparoscopia/métodos , Grampeamento Cirúrgico/métodos , Análise de Variância , Anastomose Cirúrgica , Animais , Constrição , Modelos Animais de Doenças , Imuno-Histoquímica , Masculino , Tamanho do Órgão , Probabilidade , Distribuição Aleatória , Medição de Risco , Sensibilidade e Especificidade , Estatísticas não Paramétricas , Suínos , Resistência à Tração , Fatores de TempoRESUMO
The mechanism by which glucocorticoids govern antiproteinuric effect in nephrotic syndrome remains unknown. Present study examined the protective role of dexamethasone (DEX) in the intracellular trafficking of nephrin under endoplasmic reticulum (ER) stress. Human embryonic kidney-293 cell line expressing a full-length human nephrin was cultured in mediums containing 5.5 or 25 mM glucose with or without DEX. The result revealed that glucose starvation evoked a rapid ER stress leading to formation of underglycosylated nephrin that was remained in the ER as a complex with calreticulin/calnexin. DEX rescued this interfered trafficking through binding to its receptor and stimulating the mitochondrial transcripts and adenosine 5' triphosphate (ATP) production, leading to synthesis of fully glycosylated nephrin. These results suggest that ER-stress in podocytes may cause alteration of nephrin N-glycosylation, which may be an underlying factor in the pathomechanism of the proteinuria in nephrotic syndrome. DEX may restore this imbalance by stimulating expression of mitochondrial genes, resulted in the production of ATP that is essential factor for proper folding machinery aided by the ER chaperones.
Assuntos
Dexametasona/farmacologia , Retículo Endoplasmático/efeitos dos fármacos , Glucocorticoides/uso terapêutico , Nefropatias/tratamento farmacológico , Proteínas de Membrana/metabolismo , Estresse Fisiológico , Trifosfato de Adenosina/análise , Transporte Biológico , Northern Blotting , Western Blotting , Linhagem Celular , Meios de Cultura/química , Retículo Endoplasmático/ultraestrutura , Citometria de Fluxo , Técnica Indireta de Fluorescência para Anticorpo , Corantes Fluorescentes , Glucose/análise , Humanos , Hidrazinas , Proteínas de Membrana/ultraestrutura , Microscopia Confocal , Testes de Precipitina , Proteínas/análiseRESUMO
AIMS: To detect quantitatively the total bacteria and Streptococcus mutans in dental plaque by real-time PCR with prbac, Sm and GTF-B primers, and to compare their presence with the prevalence of dental caries in Japanese preschool children. METHODS AND RESULTS: Human dental plaque samples were collected from the labial surfaces of the upper primary central incisors of 107 children. The dental status was recorded as dft by WHO caries diagnostic criteria. Positive dt and dft scores by the Sm or GTF-B primer were significantly higher than negative scores (P < 0.01). The proportions of Strep. mutans to the total bacteria from sound, and sound and/or filled upper primary incisors were significantly lower than those from decayed or filled, and decayed incisors, respectively (P < 0.01). CONCLUSIONS: The ratios of Strep. mutans to total bacteria in plaque detected by real-time PCR with Sm and GTF-B primers were closely associated with the prevalence of dental caries in Japanese preschool children. SIGNIFICANCE AND IMPACT OF THE STUDY: These assays may be useful for the assessment of an individual's risk of dental caries.
Assuntos
Cárie Dentária/microbiologia , Placa Dentária/microbiologia , Infecções Estreptocócicas/microbiologia , Streptococcus mutans/isolamento & purificação , Criança , Pré-Escolar , Primers do DNA , DNA Bacteriano/análise , Cárie Dentária/epidemiologia , Humanos , Incidência , Japão/epidemiologia , Reação em Cadeia da Polimerase , Infecções Estreptocócicas/epidemiologia , Streptococcus mutans/genéticaRESUMO
Recent reports of toxoplasmosis in marine mammals raise concern that cold-blooded marine animals are a potential source of Toxoplasma gondii infection. To examine the transmissibility of T. gondii to fish, we observed the development of T. gondii tachyzoites inoculated into oviduct epithelial cells of goldfish (Carassius auratus) microscopically in vitro. Further, the survival period of tachyzoites inoculated into goldfish muscle was bioassayed in mice and through PCR analysis. In cell cultures at 37 C, both RH and Beverley strains of T. gondii tachyzoites had penetrated into cells at 6 hr post inoculation, and were multiplying. In cell cultures at 33 C, many tachyzoites of both strains attached to the host cells, but no intracellular tachyzoites were observed at 24 hr post inoculation. In the T. gondii inoculated goldfish kept at 33 C, tachyzoite DNA was detected in the inoculated region on day 3, but not on day 7. When inoculated goldfish were kept at 37 C, live tachyzoites were seen at the inoculation site on day 3, but not on day 7. These results suggest that T. gondii does not persist in fish.
Assuntos
Doenças dos Peixes/parasitologia , Carpa Dourada/parasitologia , Toxoplasma/fisiologia , Toxoplasmose Animal/transmissão , Animais , Bioensaio , Células Cultivadas , Vetores de Doenças/classificação , Golfinhos/parasitologia , Células Epiteliais/parasitologia , Feminino , Doenças dos Peixes/transmissão , Camundongos , Camundongos Endogâmicos ICR , Oviductos/citologia , Oviductos/parasitologia , Temperatura , Toxoplasmose Animal/parasitologiaRESUMO
OBJECTIVE: To assess endometrial receptivity in terms of endometrial tissue blood flow (ETBF) measured hysterofiberscopically by laser blood-flowmetry, and to examine the technique's effectiveness in an in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) program. DESIGN: A prospective clinical study. SETTING(S): IVF program in a university hospital. PATIENT(S): A total of 75 infertile women with normal menstrual cycles undergoing IVF/ICSI. INTERVENTION(S): ETBF, conventional ultrasonographic, endocrinologic, and histologic parameters for receptivity and immunoreactivity for vascular endothelial growth factor (VEGF) in endometrium were assessed between days 4 and 6 of the luteal phase in a spontaneous menstrual cycle. Then all patients underwent IVF/ICSI. MAIN OUTCOME MEASURE(S): Achievement of clinical pregnancy by IVF/ICSI. RESULT(S): ETBF, VEGF expression, and the number of embryos were significantly higher in the women who became pregnant than in those who did not. By stepwise multiple logistic regression, significant predictors of pregnancy were the number of embryos and ETBF but not conventional receptivity markers. The rate of pregnancy was significantly higher in women with ETBF values of at least 29 mL/min per 100 grams of tissue than in women with lower values (42 vs. 15% in 36 and 39 women, respectively). ETBF was significantly greater in morphologically normal than abnormal uteri. In normal uteri, ETBF was greatest in the fundus. Correspondingly, in normal uteri 85% of gestational sacs were implanted in the fundus. CONCLUSION(S): ETBF is superior to conventional parameters for determining endometrial receptivity for implantation.
Assuntos
Implantação do Embrião/fisiologia , Endométrio/irrigação sanguínea , Fertilização in vitro/métodos , Adulto , Biópsia , Velocidade do Fluxo Sanguíneo/fisiologia , Endométrio/diagnóstico por imagem , Fatores de Crescimento Endotelial/análise , Estradiol/sangue , Feminino , Humanos , Histeroscopia , Imuno-Histoquímica , Linfocinas/análise , Masculino , Microscopia de Fluorescência , Gravidez , Progesterona/sangue , Prolactina/sangue , Estudos Prospectivos , Análise de Regressão , Ultrassonografia Doppler em Cores , Fator A de Crescimento do Endotélio Vascular , Fatores de Crescimento do Endotélio VascularAssuntos
Guerra Nuclear , Plutônio/efeitos adversos , Plutônio/farmacocinética , Cinza Radioativa/efeitos adversos , Cinza Radioativa/análise , Radioisótopos de Césio/análise , Coleta de Dados , Monitoramento Ambiental , Humanos , Cooperação Internacional , Japão , Plutônio/análise , Saúde Pública , Apoio à Pesquisa como AssuntoRESUMO
While considering the geological disposal of radioactive wastes, the behaviour of the radionuclide Np and its daughter element Pa was investigated in the presence of a mixture of anaerobic bacteria (MAB). Originally, MAB were used for the treatment of pulp and paper wastewater. The interaction between radionuclides and bacteria was evaluated by determining distribution coefficients (Kd) over 10 days and at 5 degrees C and 35 degrees C. Kd for Np at 35 degrees C after 5 days had a low value around 10(-2) After 10 days, however, Kd was > 100-fold higher. On the other hand, Kd at 5 degrees C was low (10(-2)) throughout, without any significant increase over time. The interaction between Pa and MAB was found to be stronger than that for Np, with Kd for Pa about 100 times higher. The Kd was controlled by some basic factors, the activity of MAB, the complexing capacity of MAB, and the chemical conditions in the solution such as pH and Eh.
Assuntos
Bactérias Anaeróbias/metabolismo , Netúnio/farmacocinética , Protoactínio/farmacocinética , Adsorção , Concentração de Íons de Hidrogênio , Cinética , Resíduos Radioativos , Temperatura , Microbiologia da Água , Poluentes Radioativos da Água/farmacocinéticaRESUMO
Although tumor necrosis factor receptor-associated factor 6 (TRAF6) is required in receptor activator of NF-kappaB-receptor activator of NF-kappaB ligand (RANK-RANKL) signaling for osteoclastogenesis, it has remained unclear whether TRAF6 is crucial in tumor necrosis factor alpha (TNF-alpha)-induced osteoclastogenesis. We examined TRAF6 function in the TNF-alpha-induced osteoclastogenesis by using osteoclast progenitor cells from TRAF6-deficient mice. The results indicated that TNF-alpha did not effectively induce osteoclast differentiation from osteoclast progenitor cells derived from these mice into mature multinucleated osteoclasts, although c-jun N-terminal kinase (JNK) and TNF-alpha activation was observed in osteoclast progenitor cells. Thus, we have provided the first evidence showing that TRAF6 is involved in TNF-alpha-induced osteoclastogenesis.
Assuntos
Osteoblastos/citologia , Osteogênese/fisiologia , Proteínas/metabolismo , Receptores do Fator de Necrose Tumoral/metabolismo , Transdução de Sinais , Células-Tronco/citologia , Fator de Necrose Tumoral alfa/metabolismo , Animais , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Proteínas Quinases JNK Ativadas por Mitógeno , Camundongos , Camundongos Knockout , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/metabolismo , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Proteínas/genética , Proteínas/fisiologia , Células-Tronco/efeitos dos fármacos , Células-Tronco/metabolismo , Fator 6 Associado a Receptor de TNF , Fator de Necrose Tumoral alfa/farmacologia , Regulação para CimaRESUMO
Osteoblasts are derived from mesenchymal/stromal cells in bone marrow, and gain the ability to support osteoclastogenesis during differentiation though the expression of receptor activator of NF-kappaB ligand (RANKL) and macrophage-colony stimulating factor (M-CSF). However, the properties (differentiation stage and expression of osteoblast marker genes) of stromal or osteoblastic cells that have the capacity to support osteoclast differentiation are unclear. Therefore, we sought to establish and characterize bone marrow-derived stromal cell lines (TSB) from temperature-sensitive SV40 T-antigen transgenic mice to define them at the clonal level. Of the 24 randomly selected cell lines, only 2 cell lines, TSB13 and TSB20, could support osteoclast differentiation in the presence of 1alpha,25(OH)(2)D(3). In both cell lines, RANKL mRNA was induced and osteoprotegerin (OPG) mRNA was decreased in response to treatment with 1alpha,25(OH)(2)D(3) for 2 days. Other RNA expression analyses of osteoblast-specific marker genes demonstrated the following characteristics of TSB13 and TSB20: (1) alkaline phosphatase (ALP) and type I collagen genes are expressed; (2) osteocalcin and osteopontin genes are expressed at low levels, and their expression levels are upregulated after induction of differentiation by a temperature shift from 33 degrees C to 37 degrees C, or 1alpha,25(OH)(2)D(3) treatment. Consequently, the long-term culture of TSB13 and TSB20 cell lines strongly stimulated osteocalcin expression and effectively induced calcified nodule formation in the presence of phosphate. The results suggest that the supportive cells for osteoclastogenesis are restricted to a specialized population of bone marrow stromal cells, and the high ratio of RANKL vs. OPG expression found in this population after 1alpha,25(OH)(2)D(3) treatment might be a general property of osteoclast-supporting cells.
Assuntos
Antígenos Transformantes de Poliomavirus/genética , Células da Medula Óssea/citologia , Osteoclastos/citologia , Animais , Biomarcadores , Calcificação Fisiológica/fisiologia , Proteínas de Transporte/genética , Diferenciação Celular , Linhagem Celular Transformada , Expressão Gênica , Masculino , Glicoproteínas de Membrana/genética , Camundongos , Osteoblastos/citologia , Ligante RANK , Receptor Ativador de Fator Nuclear kappa-B , Células Estromais/citologia , TemperaturaRESUMO
The migration and adhesion of osteoblasts requires several classical cadherins. Cadherin-11, one of the classical cadherins, was expressed in mouse osteoblasts in skull bone and femur, revealed by immunohistochemistry. To elucidate the function of cadherin-11 in osteoblastogenesis, cadherin-11 null mutant mice were investigated. Although apparently normal at birth, Alizarin red staining of null mutant mice showed a reduced calcified area at the frontal suture that caused a round-shaped calvaria with increasing animal age to 3 months. Consequently, there was a reduction in bone density at the femoral metaphyses and the diploë of calvaria in null mutant mice. In the in vitro culture of newborn calvarial cells, the calcified area of mutant cells was smaller than those derived from wild-type littermates. These results show that absence of cadherin-11 leads to reduced bone density in some parts of skeletons including calvaria and long bone metaphyses, and thus suggest that cadherin-11 plays roles in the regulation of osteoblast differentiation and in the mineralization of the osteoid matrix.
Assuntos
Densidade Óssea/genética , Caderinas/genética , Caderinas/metabolismo , Fêmur/anormalidades , Deleção de Genes , Crânio/anormalidades , Animais , Animais Recém-Nascidos , Western Blotting , Diferenciação Celular/genética , Células Cultivadas , Feminino , Fêmur/fisiologia , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Knockout , Mutagênese Sítio-Dirigida/genética , Osteoblastos/citologia , Osteoblastos/metabolismo , Osteogênese/genética , Fenótipo , Reação em Cadeia da Polimerase , Crânio/fisiologia , Tíbia/anormalidades , Tíbia/fisiologia , Tomógrafos ComputadorizadosRESUMO
The KI and KII sites play a crucial role in kappa-chain gene rearrangement, which was investigated in mice deficient for these sites. Previously, we found that Pax-5 can bind to the KI and KII sites; however, the function of Pax-5 in kappa-chain gene rearrangement has not been investigated. Here, we have used an in vitro culture system in which differentiation from pre-B cells to immature B cells is induced by removing IL-7. We showed that, after the induction of differentiation, Pax-5 dissociated from the KI and KII revealed by EMSA analyses, and this dissociation occurred specifically at the KI and KII sites, but not at the Pax-5 binding site, in the CD19 promoter because of a lower binding affinity of Pax-5 for the KI and KII sites. During differentiation induced by removing IL-7, the underphosphorylated form of retinoblastoma preferentially associated with Pax-5, which caused dissociation of Pax-5 from KI and KII sites. These results suggest that the dissociation of Pax-5 from the KI and KII sites is important in the induction of kappa-chain gene rearrangement.
Assuntos
Proteínas de Ligação a DNA/metabolismo , Rearranjo Gênico de Cadeia Leve de Linfócito B , Cadeias kappa de Imunoglobulina/genética , Cadeias kappa de Imunoglobulina/metabolismo , Proteína do Retinoblastoma/metabolismo , Fatores de Transcrição/metabolismo , Motivos de Aminoácidos , Animais , Células da Medula Óssea/citologia , Células da Medula Óssea/imunologia , Células da Medula Óssea/metabolismo , Diferenciação Celular/genética , Diferenciação Celular/imunologia , Linhagem Celular , Células Cultivadas , Células-Tronco Hematopoéticas/citologia , Células-Tronco Hematopoéticas/imunologia , Células-Tronco Hematopoéticas/metabolismo , Glicoproteínas de Membrana/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Fator de Transcrição PAX5 , Fosforilação , Receptores de Células Precursoras de Linfócitos B , Ligação Proteica/genética , Ligação Proteica/imunologia , Receptores de Antígenos de Linfócitos B/metabolismo , Proteína do Retinoblastoma/imunologiaRESUMO
Although expression of cutaneous lymphocyte-associated antigen (CLA) is thought to be a specific marker of skin "homing" T cells, it has become clear that CLA is a good marker for high levels of fucosyltransferase VII (Fuc-TVII) activity but does not necessarily represent the epitope required for binding to E-selectin (Wagers et al, 1996, 1997). Therefore, expression of Fuc-TVII is an attractive candidate for identifying skin "homing" T cells. However, analyses of Fuc-TVII expression in human T cells have been performed only at the mRNA level (Nakayama et al, 2000) because of the lack of mAB: In this study Fuc-TVII was for the first time visualized in individual cells by using a novel mAb that we developed. Double immunofluorescence and immunohistochemistry demonstrated the coexpression of Fuc-TVII and CLA in cell lines in which Fuc-TVII mRNA was shown to be expressed at high levels: whereas CLA expression was seen in the cell membrane, Fuc-TVII was identified in a supra- or perinuclear location. Cytoplasmic Fuc-TVII expression was also detectable in both CD4(+) and CD8(+) T cells purified from peripheral blood. Fuc-TVII was also expressed at high levels in many CLA(+) T cells infiltrating the skin. In these peripheral T cells, unlike in cell lines, cytoplasmic expression of Fuc-TVII was not always associated with surface CLA expression. This mAb would serve as a valuable tool for selectively identifying a novel subset of skin "homing" T cells that are not detected by the conventional method because of the lack of CLA expression.
Assuntos
Fucosiltransferases/análise , Imuno-Histoquímica/métodos , Glicoproteínas de Membrana/análise , Pele/imunologia , Linfócitos T/enzimologia , Anticorpos Monoclonais/imunologia , Antígenos de Diferenciação de Linfócitos T , Antígenos de Neoplasias/análise , Linhagem Celular , Fucosiltransferases/imunologia , Humanos , Glicoproteínas de Membrana/imunologia , Receptores de Retorno de Linfócitos/imunologiaRESUMO
Osteoblasts are derived originally from pluripotent mesenchymal stem cells on migration into the bone matrix. To elucidate the contribution of classical cadherins in this differentiation pathway, we developed a new protocol for their analysis and studied their specific expressions in various cell lines of the mesenchymal lineage, including osteoblasts. N-cadherin was expressed constitutively in all cell lines examined except an osteocyte-like cell line whereas cadherin-11 was expressed selectively in preosteoblast and preadipocyte cell lines. P-cadherin also was expressed in primary cultures of calvarial cells and mature osteoblasts at a relatively low level compared with N-cadherin and cadherin-11. M-cadherin was expressed only in a premyoblast cell line. We observed the transition of cadherin expression from M-cadherin to cadherin-11 in the premyoblast cell line when osteogenic differentiation was induced by treatment with bone morphogenetic protein 2 (BMP-2), while the expression of N-cadherin remained unchanged. In contrast, when a preadipocyte cell line, which shows a similar pattern of cadherin expression to osteoblasts, was induced to undergo adipogenic differentiation, the expression of N-cadherin and cadherin-11 was decreased. These observations characterize the cadherin expression profile of mesenchymal lineage cells, especially osteoblasts, which regularly express cadherin-11. Cadherin-11 may affect cell sorting, alignment, and separation through differentiation.
Assuntos
Caderinas/metabolismo , Diferenciação Celular , Mesoderma/citologia , Osteoblastos/metabolismo , Adipócitos/metabolismo , Animais , Sequência de Bases , Proteínas Morfogenéticas Ósseas/farmacologia , Linhagem da Célula , Primers do DNA , Camundongos , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Proteínas Recombinantes/farmacologia , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
CD9, a member of the tetraspan family of proteins, is highly expressed on macrophages. Although a clear function for the molecule has yet to be described, we have found that the anti-CD9 mAb activates mouse macrophages. The rat anti-CD9 mAb, KMC8.8, but not the F(ab')(2), induced tyrosine phosphorylation of proteins including syk and cbl and induced cell aggregation in the mouse macrophage cell line, J774, suggesting that co-cross-linking of CD9 and Fc gamma R was required for the signal. Co-cross-linking of CD9-Fc gamma R with KMC8.8 on macrophages from three different FcR-deficient mice, FcR gamma-chain(-/-), Fc gamma RIIB(-/-), and Fc gamma RIII(-/-), revealed that Fc gamma RIII is specific and crucial for syk phosphorylation. Although both KMC8.8 and the anti-Fc gamma RIIB/III mAb, 2.4G2, evoked similar phosphorylation patterns, only KMC8.8 induced cell aggregation. Additionally, KMC8.8 treatment led to reduce levels of TNF-alpha production and p42/44 extracellular signal-related kinase phosphorylation relative to 2.4G2 stimulation. Immunofluorescence staining showed that co-cross-linking of CD9-Fc gamma R with KMC8.8 induced filopodium extension before cell aggregation, which was followed by simultaneous colocalization of CD9, Fc gamma RIIB/III, Mac-1, ICAM-1, and F-actin at the cell-cell adhesion site. Moreover, KMC8.8 treatment of Fc gamma R-deficient macrophages revealed that the colocalization of CD9, Fc gamma RIII, Mac-1, and F-actin requires co-cross-linking of CD9-Fc gamma RIII, whereas co-cross-linking of CD9-Fc gamma RIIB induced the colocalization of only CD9 and Fc gamma RIIB. Our results demonstrate that co-cross-linking of CD9 and Fc gamma Rs activates macrophages; therefore, CD9 may collaborate with FcRs functioning in infection and inflammation on macrophages.
Assuntos
Antígenos CD/fisiologia , Macrófagos/imunologia , Macrófagos/metabolismo , Glicoproteínas de Membrana , Receptores de IgG/metabolismo , Actinas/metabolismo , Animais , Anticorpos Monoclonais/metabolismo , Anticorpos Monoclonais/farmacologia , Antígenos CD/imunologia , Antígenos CD/metabolismo , Adesão Celular/imunologia , Agregação Celular/imunologia , Linhagem Celular , Tamanho Celular/imunologia , Colesterol/metabolismo , Detergentes , Glicolipídeos/metabolismo , Soros Imunes/metabolismo , Molécula 1 de Adesão Intercelular/metabolismo , Antígeno de Macrófago 1/metabolismo , Macrófagos/citologia , Lipídeos de Membrana/metabolismo , Camundongos , Fosforilação , Fosfotirosina/metabolismo , Pseudópodes/imunologia , Receptores de IgG/imunologia , Receptores de IgG/fisiologia , Transdução de Sinais/imunologia , Solubilidade , Tetraspanina 29 , Fator de Necrose Tumoral alfa/biossínteseRESUMO
Fading is one of the major obstacles to reliable observation in fluorescence microscopy. Using a confocal laser scanning microscope (CLSM) coupled to a computer, we quantitatively measured fading of fluorescence to formulate an equation, evaluated the anti-fading ability of several anti-fading media, and restored the faded images to the original level according to this equation. NIH 3T3 cells were stained with fluorescein isothiocyanate (FITC)-phalloidin, mounted with several commercial and homemade anti-fade media, and observed with CLSM under repeated illumination. With any mounting medium, attenuation of fluorescence intensity at a certain pixel occurred stepwise and the decrease was proportional to the intensity of the previous scan. From these results, we formulated an equation that has three coefficients: anti-fading factor (A), indicating the ability to retard fading; fluorescent intensity at the first scan (EM(1)); and background fluorescence (B). The fluorescent intensity at a certain point following nth scan is given as EM(n) = EM(1) * A ((n-1)). This equation was available for restoring faded images to their original states, even after the image had faded to only 60% of its original intensity.
Assuntos
Aumento da Imagem , Microscopia Confocal/métodos , Células 3T3 , Algoritmos , Animais , Fluoresceína-5-Isotiocianato , Corantes Fluorescentes , Indicadores e Reagentes , Camundongos , FaloidinaRESUMO
Trace amounts of heavy metals in the ice cores from Canadian Arctic were analyzed using inductively coupled plasma mass spectrometry (ICP-MS). A custom made plastic device and ceramic knives were used to remove the contamination on the ice core surface. Ice cores could be broken into small sections (2-3 cm thick) after decontamination with the plastic device and ceramic knives. High-resolution depth profiles of various elements, i.e. As, Cd, Co, Cu, Ni, Pb, Zn and U, were thus attained. Concentrations in 518 ice core samples range from 0.1 (U) to 673.3 (Zn) pg g(-1).
RESUMO
Involvement of the central nervous system is uncommon in progressive systemic sclerosis, with only 2 reported cases associated with intracerebral hemorrhage detected by neuroimaging. A 55-year-old woman with a 10-year history of scleroderma presented with left occipital lobe hemorrhage manifesting as headache and vomiting. She had no signs of hypertension, diabetes mellitus and hyperlipidemia. CT and MRI, on admission, showed left occipital lobe hemorrhage with ventricular rupture and acute left subdural hematoma. Serial cerebral angiography was performed on day 0, day 7 and day 14, and found no evidence of aneurysm, arteriovenous multiformation or tumor stain in the left occipital lobe. However, the bilateral anterior cerebral arteries showed increasing segmental narrowing suggestive of vasculitis. Histological examination of a section from the brain cortex adjacent to the hemorrhage revealed no evidence of vasculitis, fibrinoid degeneration or amyloid deposition. Focal vasculitis may have occurred secondary to the homorrhagic lesion.