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1.
J Vis Exp ; (42)2010 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-20972388

RESUMO

The sense of taste is critical for human life. It informs the body about the quality of food that will be potentially ingested and stimulates metabolic processes that prepare the alimentary canal for digestion. Steady progress is being made towards understanding the early biochemical and molecular events underlying taste transduction (for a review, Breslin and Spector, 2008). However, progress to date has largely resulted from animal models. Yet, since marked differences in receptor specificity and receptor density vary among species, human taste transduction will only be understood by using human taste tissue. Here we describe a biopsy technique to collect human fungiform papillae, visible as rounded pink anterior structures, about 0.5 mm in diameter that contain taste buds. These biopsied papillae are used for several purposes including the isolation of viable taste bud cells, in situ hybridization, immunohistochemistry and, through techniques of molecular biology, the identification of taste-specific novel proteins.


Assuntos
Biópsia/métodos , Papilas Gustativas/anatomia & histologia , Língua/anatomia & histologia , Humanos , Papilas Gustativas/cirurgia , Língua/cirurgia
2.
Chem Senses ; 35(6): 501-9, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20457570

RESUMO

A healthy taste system is important to the maintenance of nutrition and overall quality of life, and taste disorders are associated with many inflammatory states. We previously determined the immune cells in normal human gustatory tissue; they are predominantly dendritic cells and CD4 T cells with a few macrophages and B lymphocytes present. There are, however, few reports of the subtypes of resident lymphocytes in or near taste tissues. The present study further characterized the distribution and population of the major subtypes of T cells in situ within biopsies of healthy human fungiform papillae (FP). Immunohistochemical analyses indicated that T-helper (Th)1 cells (CCR5+) were more predominant in FP than Th2 T cells (CCR4+). CD45RO+ memory T cells were the principal T cells in gustatory tissue, whereas CD45RA+ naive T cells were uncommon. Regarding subcompartments of the tissue, most intraepithelial lymphocytes of FPs were gamma/delta T cells, whereas the major subtype of lymphocytes in the lamina propria were alpha/beta T cells. Regulatory T cells that express CTLA-4 (CD152) and interleukin-2 receptors (IL-2R, CD25) were found at low levels in FP. The T cells stand ready to respond to inflammatory and infectious insults and may play a role in the taste alterations observed during acute and chronic inflammatory states.


Assuntos
Memória Imunológica , Subpopulações de Linfócitos T/imunologia , Papilas Gustativas/imunologia , Células Th1/imunologia , Adulto , Antígenos CD/metabolismo , Antígeno CTLA-4 , Feminino , Humanos , Imuno-Histoquímica , Subunidade alfa de Receptor de Interleucina-2/metabolismo , Antígenos Comuns de Leucócito/metabolismo , Masculino , Receptores CCR4/metabolismo , Receptores CCR5/metabolismo , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/metabolismo , Células Th1/metabolismo , Células Th2/imunologia , Células Th2/metabolismo , Língua/citologia
3.
PLoS One ; 4(10): e7347, 2009 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-19812697

RESUMO

BACKGROUND: The perception of sour taste in humans is incompletely understood at the receptor cell level. We report here on two patients with an acquired sour ageusia. Each patient was unresponsive to sour stimuli, but both showed normal responses to bitter, sweet, and salty stimuli. METHODS AND FINDINGS: Lingual fungiform papillae, containing taste cells, were obtained by biopsy from the two patients, and from three sour-normal individuals, and analyzed by RT-PCR. The following transcripts were undetectable in the patients, even after 50 cycles of amplification, but readily detectable in the sour-normal subjects: acid sensing ion channels (ASICs) 1a, 1beta, 2a, 2b, and 3; and polycystic kidney disease (PKD) channels PKD1L3 and PKD2L1. Patients and sour-normals expressed the taste-related phospholipase C-beta2, the delta-subunit of epithelial sodium channel (ENaC) and the bitter receptor T2R14, as well as beta-actin. Genomic analysis of one patient, using buccal tissue, did not show absence of the genes for ASIC1a and PKD2L1. Immunohistochemistry of fungiform papillae from sour-normal subjects revealed labeling of taste bud cells by antibodies to ASICs 1a and 1beta, PKD2L1, phospholipase C-beta2, and delta-ENaC. An antibody to PKD1L3 labeled tissue outside taste bud cells. CONCLUSIONS: These data suggest a role for ASICs and PKDs in human sour perception. This is the first report of sour ageusia in humans, and the very existence of such individuals ("natural knockouts") suggests a cell lineage for sour that is independent of the other taste modalities.


Assuntos
Regulação da Expressão Gênica , Papilas Gustativas/metabolismo , Paladar/fisiologia , Língua/metabolismo , Canais Iônicos Sensíveis a Ácido , Idoso de 80 Anos ou mais , Biópsia , Canais de Cálcio/biossíntese , Canais Epiteliais de Sódio/biossíntese , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Proteínas do Tecido Nervoso/biossíntese , Fosfolipase C beta/biossíntese , Receptores de Superfície Celular/biossíntese , Receptores Acoplados a Proteínas G/biossíntese , Canais de Sódio/biossíntese , Paladar/genética
4.
Brain Behav Immun ; 23(6): 760-6, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19268521

RESUMO

Taste loss or alterations can seriously impact health and quality of life due to the resulting negative influence on eating habits and nutrition. Infection and inflammation are thought to be some of the most common causes of taste perception disorders. Supporting this view, neuro-immune interactions in the peripheral gustatory system have been identified, underlying the importance of this tissue in mucosal immunity, but we have little understanding of how these interactions influence taste perception directly or indirectly. This limited understanding is evident by the lack of even a basic knowledge of the resident immune cell populations in or near taste tissues. The present study characterized the distribution and population of the major immune cells and their subsets in healthy human anterior, lingual, fungiform papillae (FP) using immunohistochemistry. Dendritic cells (DCs) were the predominant innate immune cells in this tissue, including four subtypes: CD11c(+) DCs, DC-SIGN+ immature DCs, CD83(+) mature DCs, and CD1a(+) DCs (Langerhans cells). While most DCs were localized beneath the lamina propria and only moderately in the epithelium, CD1a(+) Langerhans cells were exclusively present within the epithelium and not in sub-strata. A small number of macrophages were observed. T lymphocytes were present throughout the FP with CD4(+) T cells more prevalent than CD8(+) T cells. Very few CD19(+) B lymphocytes were detected. The results show that DCs, macrophages, and T lymphocytes are the constitutive guardians of human FP taste tissue, with DCs and CD4 T cells being dominant, while B lymphocytes are rare under normal, healthy conditions. These observations provide a basic anatomical foundation for the immune response in the healthy human tongue as a basis for subsequent disease-related studies, but none of the present data indicate that the immune cell populations identified are, in fact, altered in individuals with abnormal taste perception.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Células Dendríticas/imunologia , Imunidade Celular/imunologia , Papilas Gustativas/imunologia , Paladar/imunologia , Adulto , Antígenos CD/imunologia , Antígeno CD11c/imunologia , Contagem de Células , Feminino , Genes MHC da Classe II/genética , Humanos , Processamento de Imagem Assistida por Computador , Imunoglobulinas/imunologia , Imuno-Histoquímica , Células de Langerhans/imunologia , Macrófagos/imunologia , Masculino , Glicoproteínas de Membrana/imunologia , Papilas Gustativas/fisiologia , Língua/citologia , Antígeno CD83
5.
Br J Oral Maxillofac Surg ; 42(4): 348-50, 2004 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-15225956

RESUMO

The purpose of this paper is to present a case report of a benign mesenchymoma involving the lip. A brief review of relevant literature is presented.


Assuntos
Neoplasias Labiais/patologia , Mesenquimoma/patologia , Humanos , Neoplasias Labiais/cirurgia , Masculino , Mesenquimoma/cirurgia , Pessoa de Meia-Idade
6.
Chem Senses ; 29(1): 13-23, 2004 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-14752036

RESUMO

Various genes related to early events in human gustation have recently been discovered, yet a thorough understanding of taste transduction is hampered by gaps in our knowledge of the signaling chain. As a first step toward gaining additional insight, the expression specificity of genes in human taste tissue needs to be determined. To this end, a fungiform papillae cDNA library has been generated and analyzed. For validation of the library, taste-related gene probes were used to detect known molecules. Subsequently, DNA sequence analysis was performed to identify further candidates. Of 987 clones sequenced, clustering results in 288 contigs. Comparison of these contigs with genomic databases reveals that 207 contigs (71.9%) match known genes, 16 (5.6%) match hypothetical genes, eight (2.8%) match repetitive sequences and 57 (19.8%) have no or low similarity to annotated genes. The results indicate that despite a high level of redundancy, this human fungiform cDNA library contains specific taste markers and is valuable for investigation of both known and novel taste-related genes.


Assuntos
Biblioteca Gênica , Papilas Gustativas , Paladar/genética , Biologia Computacional , Mapeamento de Sequências Contíguas , Bases de Dados de Ácidos Nucleicos , Humanos , Proteínas/classificação , Proteínas/genética , Análise de Sequência , Transdução de Sinais/genética
7.
Head Neck ; 25(11): 911-21, 2003 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-14603451

RESUMO

BACKGROUND: The clinical utility of tolonium chloride rinse was compared with unaided visual examination alone in the diagnosis of oral carcinoma in patients previously treated for carcinoma of the upper aerodigestive tract. METHODS: A total of 668 patients were enrolled in this multicenter study. At each site, an oral clinical visual examination was completed by one investigator followed by tolonium chloride rinse and examination by a second investigator blinded to the examination findings of the first investigator. If a lesion was considered suspicious (urgent biopsy required at the first visit), the lesion was biopsied after tolonium chloride rinsing. Patients with lesions characterized at the first visit as not suspicious (biopsy not urgent) or that stained with tolonium chloride were asked to return for a second visit. At the second visit, any residual lesion or lesions that retained tolonium chloride were biopsied. RESULTS: A total of 96 biopsies was performed in 81 of the 668 patients (12.1%), of which 30 (31.3%) were diagnosed histologically as carcinoma/carcinoma in situ (CIS) and the remainder as inflammation (31.3%), keratosis (26.6%), dysplasia (21.9%), ulcer (2.1%), other (3.1%), or no abnormality (1.0%). Of the 30 lesions with the diagnosis of carcinoma/CIS, 12 (sensitivity 40.0%) were considered to be clinically suspicious (CS+), whereas 29 (sensitivity 96.7%) retained tolonium chloride (p =.0002). The predictive values of a positive test for clinical examination and tolonium chloride staining were similar (36.4% vs 32.6%; p =.5871), indicating that the greater sensitivity of tolonium chloride was not associated with an excessive number of unnecessary biopsies (false positives). CONCLUSIONS: Tolonium chloride rinse is more sensitive than clinical examination alone in detecting lesions that might be found on biopsy to be carcinoma or CIS. The increased sensitivity is largely attributed to lesions that stain but were not detected clinically on visual examination.


Assuntos
Corantes , Neoplasias de Cabeça e Pescoço/diagnóstico , Neoplasias Laríngeas/diagnóstico , Neoplasias Bucais/diagnóstico , Segunda Neoplasia Primária/diagnóstico , Cloreto de Tolônio , Idoso , Feminino , Neoplasias de Cabeça e Pescoço/patologia , Humanos , Leucoplasia Oral/diagnóstico , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/diagnóstico , Segunda Neoplasia Primária/patologia , Neoplasias Palatinas/diagnóstico , Valor Preditivo dos Testes , Neoplasias da Língua/diagnóstico , Neoplasias Tonsilares/diagnóstico
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