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1.
J Am Dent Assoc ; 154(12): 1058-1066.e4, 2023 12.
Artigo em Inglês | MEDLINE | ID: mdl-37777935

RESUMO

BACKGROUND: Local anesthesia is an essential component of dentistry, but there is limited quantifiable understanding of what techniques and local anesthetic solutions are used by practicing dentists. Use of the local anesthetic articaine has been highly debated in dentistry regarding its efficacy and risks for paresthesia. The aims of this study were to expand the knowledge of local anesthesia practices of dentists in the United States through a large-scale survey and associate potential influencing factors regarding articaine use specifically. METHODS: The 23-item survey was sent to 10,340 practicing dentists in the United States, gathering demographic data and local anesthesia approaches and concerns. Statistical analysis consisted of descriptive, bivariate, and multivariate logistic regression analyses. RESULTS: A total of 1,128 dentists completed the survey. Previous experience with articaine was reported by 97.6% of respondents, with 3.3% no longer using articaine. Sixty percent of respondents indicated using articaine for most local anesthetic injections administered. Multivariable regression analysis found those reporting to use articaine for all local anesthetic injections involving vasoconstrictors were more likely to be male (odds ratio, 1.59; P = .002) or general dentists (odds ratio, 1.63; P < .001). CONCLUSIONS: Articaine has a perceived benefit to practitioners as most respondents reported using articaine as their primary local anesthetic. A practitioner's sex and type were found to affect the profile of use of articaine. PRACTICAL IMPLICATIONS: Assembling evidenced-based local anesthesia practices would be beneficial to ensure US practitioners are more standardized in administering local anesthetics, particularly articaine, in the safest and most efficacious way.


Assuntos
Anestesia Dentária , Carticaína , Masculino , Humanos , Estados Unidos , Feminino , Anestésicos Locais , Anestesia Dentária/métodos , Anestesia Local/métodos , Inquéritos e Questionários , Lidocaína , Método Duplo-Cego
2.
J Histochem Cytochem ; 70(3): 211-223, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-34994225

RESUMO

Multiplex immunofluorescence (mIF) is an effective technique for the maximal visualization of multiple target proteins in situ. This powerful tool is mainly limited by the spectral overlap of the currently available synthetic fluorescent dyes. The fluorescence excitation wavelengths ranging between 405 and 488 nm are rarely used in mIF imaging and serve as a logical additional slot for a fluorescent probe. In the present study, we demonstrate that the addition of 2,3,4,5,6-pentafluoroaniline to Atto 465 NHS ester, creating Atto 465-pentafluoroaniline (Atto 465-p), generates a bright nuclear stain in the violet-blue region of the visible spectrum. This allows the 405 nm excitation and emission, classically used for nuclear counterstains, to be used for the detection of another target protein. This increases the flexibility of the mIF panel and, with appropriate staining and microscopy, enables the quantitative analysis of at least six targets in one tissue section. (J Histochem Cytochem XX: XXX-XXX, XXXX).


Assuntos
Núcleo Celular/química , Proflavina/análogos & derivados , Compostos de Anilina/química , Animais , Feminino , Imunofluorescência , Corantes Fluorescentes/química , Fluorbenzenos/química , Fluorocarbonos/química , Histocitoquímica , Camundongos , Camundongos Endogâmicos BALB C , Proflavina/análise
3.
Proc Natl Acad Sci U S A ; 118(3)2021 01 19.
Artigo em Inglês | MEDLINE | ID: mdl-33446503

RESUMO

Almost 150 papers about brain lymphatics have been published in the last 150 years. Recently, the information in these papers has been synthesized into a picture of central nervous system (CNS) "glymphatics," but the fine structure of lymphatic elements in the human brain based on imaging specific markers of lymphatic endothelium has not been described. We used LYVE1 and PDPN antibodies to visualize lymphatic marker-positive cells (LMPCs) in postmortem human brain samples, meninges, cavernous sinus (cavum trigeminale), and cranial nerves and bolstered our findings with a VEGFR3 antibody. LMPCs were present in the perivascular space, the walls of small and large arteries and veins, the media of large vessels along smooth muscle cell membranes, and the vascular adventitia. Lymphatic marker staining was detected in the pia mater, in the arachnoid, in venous sinuses, and among the layers of the dura mater. There were many LMPCs in the perineurium and endoneurium of cranial nerves. Soluble waste may move from the brain parenchyma via perivascular and paravascular routes to the closest subarachnoid space and then travel along the dura mater and/or cranial nerves. Particulate waste products travel along the laminae of the dura mater toward the jugular fossa, lamina cribrosa, and perineurium of the cranial nerves to enter the cervical lymphatics. CD3-positive T cells appear to be in close proximity to LMPCs in perivascular/perineural spaces throughout the brain. Both immunostaining and qPCR confirmed the presence of adhesion molecules in the CNS known to be involved in T cell migration.


Assuntos
Encéfalo/metabolismo , Sistema Linfático/metabolismo , Glicoproteínas de Membrana/metabolismo , Receptor 3 de Fatores de Crescimento do Endotélio Vascular/genética , Proteínas de Transporte Vesicular/metabolismo , Idoso , Idoso de 80 Anos ou mais , Anticorpos/imunologia , Anticorpos/isolamento & purificação , Autopsia , Encéfalo/diagnóstico por imagem , Movimento Celular/genética , Sistema Nervoso Central/imunologia , Sistema Nervoso Central/metabolismo , Dura-Máter/diagnóstico por imagem , Dura-Máter/metabolismo , Endotélio Linfático/diagnóstico por imagem , Endotélio Linfático/metabolismo , Feminino , Sistema Glinfático/metabolismo , Humanos , Imuno-Histoquímica/métodos , Sistema Linfático/diagnóstico por imagem , Vasos Linfáticos/diagnóstico por imagem , Vasos Linfáticos/metabolismo , Masculino , Glicoproteínas de Membrana/isolamento & purificação , Espaço Subaracnóideo/diagnóstico por imagem , Espaço Subaracnóideo/metabolismo , Linfócitos T/imunologia , Proteínas de Transporte Vesicular/isolamento & purificação
4.
J Clin Med ; 9(1)2020 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-31963936

RESUMO

Sarcoidosis is a devastating inflammatory disease affecting many organs, especially the lungs and lymph nodes. Bone marrow-derived mesenchymal stromal cells (MSCs) can "reprogram" various types of macrophages towards an anti-inflammatory phenotype. We wanted to determine whether alveolar macrophages from sarcoidosis subjects behave similarly by mounting an anti-inflammatory response when co-cultured with MSCs. Fifteen sarcoidosis and eight control subjects underwent bronchoscopy and bronchoalveolar lavage (BAL). Unselected BAL cells (70-94% macrophages) were isolated and cultured with and without MSCs from healthy adults. Following stimulation of the cultured cells with lipopolysaccharide, the medium was removed to measure interleukin 10 and tumor necrosis factor alpha (IL-10 and TNF-α). In two additional sarcoidosis subjects, flow cytometry was used to study intracellular cytokines and surface markers associated with alveolar macrophages to confirm the results. Unselected BAL cells from sarcoidosis subjects co-cultured with MSCs showed a reduction in TNF-α (pro-inflammatory M1) and an increase in IL-10 (anti-inflammatory M2) in 9 of 11 samples studied. Control subject samples showed few, if any, differences in cytokine production. Unselected BAL cells from two additional patients analyzed by flow cytometry confirmed a switch towards an anti-inflammatory state (i.e., M1 to M2) after co-culture with MSCs. These results suggest that, similarly to other macrophages, alveolar macrophages also respond to MSC contacts by changing towards an anti-inflammatory phenotype. Based on our results, we hypothesize that mesenchymal stromal cells applied to the airways might alleviate lung inflammation and decrease steroid need in patients with sarcoidosis.

5.
Dev Biol ; 378(2): 141-53, 2013 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-23562806

RESUMO

Deregulation of the non-receptor tyrosine kinase ACK1 (Activated Cdc42-associated kinase) correlates with poor prognosis in cancers and has been implicated in promoting metastasis. To further understand its in vivo function, we have characterized the developmental defects of a null mutation in Drosophila Ack, which bears a high degree of sequence similarity to mammalian ACK1 but lacks a CRIB domain. We show that Ack, while not essential for viability, is critical for sperm formation. This function depends on Ack tyrosine kinase activity and is required cell autonomously in differentiating male germ cells at or after the spermatocyte stage. Ack associates predominantly with endocytic clathrin sites in spermatocytes, but disruption of Ack function has no apparent effect on clathrin localization and receptor-mediated internalization of Boss (Bride of sevenless) protein in eye discs. Instead, Ack is required for the subcellular distribution of Dock (dreadlocks), the Drosophila homolog of the SH2- and SH3-containing adaptor protein Nck. Moreover, Dock forms a complex with Ack, and the localization of Dock in male germ cells depends on its SH2 domain. Together, our results suggest that Ack-dependent tyrosine phosphorylation recruits Dock to promote sperm differentiation.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Proteínas Tirosina Quinases/metabolismo , Espermatócitos/metabolismo , Espermatogênese , Proteínas Adaptadoras de Transdução de Sinal/genética , Animais , Animais Geneticamente Modificados , Western Blotting , Diferenciação Celular/genética , Clatrina/metabolismo , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Endocitose , Proteínas do Olho/genética , Proteínas do Olho/metabolismo , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Masculino , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/metabolismo , Microscopia de Fluorescência , Proteínas do Tecido Nervoso/genética , Ligação Proteica , Proteínas Tirosina Quinases/genética , Receptores de Peptídeos/genética , Receptores de Peptídeos/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Espermatócitos/citologia , Domínios de Homologia de src/genética
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