Pérdida de tolerancia inmune en la etiología de las úlceras aftosas recidivantes (RAU) de la mucosa oral: la ruptura de la tolerancia inmunológica causaría injuria persistente en la mucosa bucal provocando las úlceras aftosas recurrentes (1ª parte)

La mucosa oral ha desarrollado un Sistema inmune único y distinto desde los primeros pasos de vida extrauterina, profundamente inmerso en un entorno nuevo y cambiante. En ese momento se produce la delección de células T autorreactivas en el timo. Se crean células nuevas, llamadas células tolerogénicas o Tregs. Con el fin de evitar la autoinmunidad y la inflamación crónica, moléculas coestimuladoras, interleucinas, factores transformantes y células dendríticas, son marcadores ayudadores a analizar. Actualmente se reconoce la cavidad oral como una región de tolerancia inmunológica. Hoy en día existen muchas definiciones de Pérdida de Tolerancia Inmune, las cuales son consultadas en esta presentación. Han sido seleccionadas las más relevantes, con el fin de asociar y explicar los tres problemas más cruciales de este proceso patológico: las úlceras aftosas recidivantes o RAU. Ellos son: dolor, vulnerabilidad y recurrencia.

Melanoma maligno da mucosa da cabeça e pescoço: casuística do IPO de Lisboa

Introdução: O melanoma maligno da mucosa (MMM) é uma doença rara com mau prognóstico. Material e Métodos: Estudo retrospetivo de 32 doentes do Instituto Português de Oncologia Francisco Gentil de Lisboa com MMM da cabeça e pescoço, no período de 1998 a 2012. Resultados: Dos 32 casos analisados a idade média foi de 70 anos. O tumor primário localizou-se na cavidade nasal e seios peri-nasais em 24 doentes e na cavidade oral em 8 casos. A maioria dos doentes (23) foi submetida a tratamento cirúrgico. Destes, 16 foram propostos para terapêutica complementar com Radioterapia. O tempo de seguimento variou de 26 dias a 10 anos. A sobrevida aos 5 anos foi de 18%. Conclusões: A maioria dos doentes apresentou um estadio avançado na altura do diagnóstico e, apesar dos tratamentos instituídos, verificou-se uma elevada mortalidade. O tratamento de escolha é a cirurgia. O papel da radioterapia continua a ser controverso.

Nuclear alterations in nasal mucosa epithelial cells of students exposed to formaldehyde

Introduction: Formaldehyde is a compound with a wide range and is commonly used in anatomy and pathology laboratories. At room temperature is quickly volatilized to a pungent and suffocating gas and its inhalation has been correlated to nuclear alterations in different tissues. We aimed to investigate whether exposure to this compound was correlated with the appearance of cytotoxic and genotoxic features in the nasal epithelial cells of students enrolled in a human anatomy course. Material and Methods: This prospective study collected periodically nasal cells from mucosa of 17 volunteers from two different undergraduate programs with different workloads of practical lessons in an anatomy laboratory, 30 and 90 hours per semester. Cells were staining according to Feulgen method and nuclear morphology was analyzed to detect possible damage. Dunn's post hoc test was used in the statistical analysis. Pearson's correlation was performed for gender, age and questionnaire responses. Results: Epithelial cells showed indicators of cytotoxicity and mutagenicity. Students with a more extensive workload in anatomy laboratory displayed a more severe profile with an increase in karyorrhexis (p < 0.05) over time. The micronucleus analysis showed difference between first and second collection (p < 0.01), although it was not maintained over the time. Students with a less extensive workload display no differences in most of cytological features. Despite karyorrhexis was present in a greater number of cells, for this group no significant difference was observed between any range. The same was observed to karyolysis and micronucleus (p > 0.05). Conclusion: Individuals exposed for short periods of time to formaldehyde are subject to the toxic action of this gas. Karyorrhexis was the most frequently observed cytotoxic feature and micronucleus showed an increase between the first time point. The patterns observed between the student's groups suggest a negative effect due to exposure time.

Fucana ativa via das Map quinases e inibe a proliferação de células de ovário de hamster chinês (CHO)

Fucan is a term used to denominate L-fucose rich sulfated polysaccharides. The fucans have been studied due their pharmacological activities like antithrombotic, antiproliferative and antioxidant. We have extracted three fucan fractions from the brown seaweed Spatoglossum schröederi. These fucans were denominated Fuc B 1, Fuc B 1.5 and Fuc B 2. The chemical analyzes show that the fucans have very similar composition as demonstrated by agarose electrophoresis gel, sugar and sulfate content. The antiproliferative effect was determined by MTT and BrdU methodologies in CHO cells. The inhibition of proliferation effect of the three fractions was about 40%. Therefore this we proceed just with the Fuc B 2 due the higher yield. There is no apoptosis indication using the anexin V/propidium iodide test. We found a cell cycle phase G1 arrest. The western blotting show that the PKC; pFAK; pERK 1/2 are activated when the cells were treated with fucans. The treatement with inhibitor of MAPK PD98059 extinguished the fucan effect. These results indicates that fucan act by the ERK pathway inducing the cell death.

Composição racial e a avaliação da reação intradérmica de Montenegro em portadores da leishmaniose cutâneo-mucosa

Em um estudo retrospectivo, indivíduos com a forma mucosa da leishmaniose foram pareados (sexo, idade e ocupação) a outros pacientes com a forma cutânea em atividade, não demonstrando-se diferença significante da freqüência nos grupos raciais. Do mesmo modo, o teste intradérmico de Montenegro não se mostrou diferente nos diversos grupos raciais, entre os 96 pares de pacientes estudados. Uma maior enduração foi observada em pacientes com o acometimento mucoso quando comparado aos cutâneos. _______________________________________________________________________________________ ABSTRACT

Pérdida de tolerancia inmune en la etiología de las úlceras aftosas recidivantes (RAU) de la mucosa oral: la ruptura de la tolerancia inmunológica causaría injuria persistente en la mucosa bucal provocando las úlceras aftosas recurrentes (2ª parte)

Se ha realizado una revisión de teorías de pérdida de tolerancia inmune publicadas tratando de encontrar aquellos conceptos modernos. Las mismas se refieren a la predisposición genética, influencia de la epigenética en la modelación de un fenotipo vulnerable, las hipótesis de higiene y de microbiota, síndrome de sensibilidad química múltiple, stress crónico, cortisol, el eje hipófisis, hipotálamo y páncreas, óxido nítrico, ataque a la membrana celular, e injuria por reperfusión. Aplicando los principios básicos de las teorías consultadas se demuestra la pérdida de homeostasis, general o parcial, que podría llegar a explicar tres características fundamentales de las úlceras recurrentes orales: dolor, vulnerabilidad y recurrencia.

Influence of factors in the oral mucosa maturation pattern: a cross-sectional study applying multivariate analyses

Aim: To evaluate the association between oral health status, socio-demographic and behavioral factors with the pattern of maturity of normal epithelial oral mucosa. Methods: Exfoliative cytology specimens were collected from 117 men from the border of the tongue and floor of the mouth on opposite sides. Cells were stained with the Papanicolaou method and classified into: anucleated, superficial cells with nuclei, intermediate and parabasal cells. Quantification was made by selecting the first 100 cells in each glass slide. Sociodemographic and behavioral variables were collected from a structured questionnaire. Oral health was analyzed by clinical examination, recording decayed, missing and filled teeth index (DMFT) and use of prostheses. Multivariable linear regression models were applied. Results: No significant differences for all studied variables influenced the pattern of maturation of the oral mucosa except for alcohol consumption. There was an increase of cell surface layers of the epithelium with the chronic use of alcohol. Conclusions: It is appropriate to use Papanicolaou cytopathological technique to analyze the maturation pattern of exposed subjects, with a strong recommendation for those who use alcohol - a risk factor for oral cancer, in which a change in the proportion of cell types is easily detected.

Contribution of transmembrane domain V amino acids to β1l-adrenoceptor activity and affinity

Introduction. There are two binding sites on the β1-adrenoceptor (AR), β1H and β1L corresponding to high and low affinity binding sites respectively, which can be activated to cause cardiostimulation. Some β-blockers that block β1AR and β2ARs can activate β1LARs at higher concentrations than those required to cause blockade. The β2AR does not form a corresponding low affinity binding site and therefore we postulated that heterologous amino acids are responsible for the formation of β1LAR. Aim. To investigate whether heterologous amino acids of transmembrane domain V (TMDV) of β1AR and β2ARs contribute to β1LAR. Methods. β1ARs, β2ARs and mutant β1ARs containing all (β1(β2TMDV)AR) or single amino acids of TMDV of the β2AR were prepared and stably expressed in Chinese Hamster Ovary cells. Concentration-effect curves for cyclicAMP accumulation were carried out for (-)-CGP12177 in the absence or presence of (-)-bupranolol. Results. The potencies (pEC50) of (-)-CGP12177 were β2AR (9.24 ± 0.14, n = 5), β1(V230I)AR (9.07 ± 0.07, n = 10), β1(β2TMDV)AR (8.86 ± 0.10, n = 15), β1(R222Q)AR (8.09 ± 0.29, n = 6), β1AR (8.00 ± 0.11, n = 11). The affinities (pKB) of (-)-bupranolol were β2AR (9.82 ± 0.52, n = 5), β1(V230I)AR (7.64 ± 0.12, n = 8), β1(β2TMV)AR (8.06 ± 0.17, n = 8), β1(R222Q)AR (7.33 ± 0.23, n = 5), β1AR (7.23 ± 0.23, n = 5). Discussion. The potency of (-)-CGP12177 was higher at β2AR than at β1AR consistent with activation through a low affinity site at the β1AR (β1LAR). The presence of V230 in β1AR accounted for the lower potency of (-)-CGP 12177. The affinity of (-)-bupranolol was lower at β1AR compared to β2AR. The presence of V230 in β1AR accounted in part for the lower affinity. In conclusion TMDV of the β1AR contributes in part to the low affinity binding site of β1AR.

Contribution of transmembrane domain V amino acids to β1l-adrenoceptor activity and affinity

There are two binding sites on the β1-adrenoceptor (AR), β1H and β1L corresponding to high and low affinity binding sites respectively, which can be activated to cause cardiostimulation (reviewed Kaumann and Molenaar, 2008). Some β-blockers that block β1AR and β2ARs can activate β1LARs at higher concentrations than those required to cause blockade. The β2AR does not form a corresponding low affinity binding site (Baker et al 2002) and therefore we postulated that heterologous amino acids are responsible for the formation of β1LAR. Our aim was to investigate whether heterologous amino acids of transmembrane domain V (TMDV) of β1AR and β2ARs contribute to β1LAR. β1ARs, β2ARs and mutant β1ARs containing all (β1(β2TMDV)AR) or single amino acids of TMDV of the β2AR were prepared and stably expressed in Chinese Hamster Ovary cells. Concentration-effect curves for cyclicAMP accumulation were carried out for (-)-CGP12177 or (-)-isoprenaline in the absence or presence of (-)-bupranolol. _______________________________________________________________________ (-)-CGP 12177 (-)-Bupranolol affinity (pKB) pEC50 vs (-)-CGP 12177 vs (-)-isoprenaline _______________________________________________________________________ β1AR 8.00 ± 0.11 (11) 7.23 ± 0.23 (5) 9.52 ± 0.28 (5) β2AR (high density) 9.24 ± 0.14 (5) 9.82 ± 0.52 (8) xPaulxxxxxxx β2AR (low density) no effect β1(β2TMV)AR 8.86 ± 0.10 (15) 8.06 ± 0.17 (8) 9.08 ± 0.22 (6) β1(V230I)AR 9.07 ± 0.07 (10) 7.64 ± 0.12 (8) 9.36 ± 0.28 (9) β1(R222Q)AR 8.09 ± 0.29 (6) 7.33 ± 0.23 (5) 9.36 ± 0.08 (6) β1(V230A)AR 7.59 ± 0.09 (6) 7.32 ± 0.24 (4) 8.62 ± 0.18 (5) _______________________________________________________________________ The potency of (-)-CGP12177 was higher at β2AR than at β1AR consistent with activation through a low affinity site at the β1AR (β1LAR) but not β2AR. The presence of V230 in β1AR accounted for the lower potency of (-)-CGP 12177. The affinity of (-)-bupranolol at β1AR and mutants was higher when determined with (-)-isoprenaline than with (-)-CGP 12177. The affinity of (-)-bupranolol determined against (-)-CGP 12177 was lower at β1AR compared to β2AR. The presence of V230 in β1AR accounted in part for the lower affinity. In conclusion V230 of the β1AR contributes in part to the low affinity binding site of β1AR. Baker JG, Hall IP, Hill SJ (2002). Pharmacological characterization of CGP12177 at the human β2-adrenoceptor. Br J Pharmacol 137, 400−408 Kaumann AJ, Molenaar P (2008) The low-affinity site of the β1-adrenoceptor and its relevance to cardiovascular pharmacology. Pharmacol Ther 118, 303-336

Chlamydial infection of immune cells : altered function and implications for disease

Chlamydia trachomatis is an obligate intracellular bacterial pathogen that infects the genital and ocular mucosa of humans, causing infections that can lead to pelvic inflammatory disease, infertility, and blinding trachoma. C. pneumoniae is a respiratory pathogen that is the cause of 12–15% of community-acquired pneumonia. Both chlamydial species were believed to be restricted to the epithelia of the genital, ocular, and respiratory mucosa; however, increasing evidence suggests that both these pathogens can be isolated from peripheral blood of both healthy individuals and patients with inflammatory conditions such as coronary artery disease and asthma. Chlamydia can also be isolated from brain tissues of patients with degenerative neurological disorders such as Alzheimer’s disease and multiple sclerosis, and also from certain lymphomas. An increasing number of in vitro studies suggest that some chlamydial species can infect immune cells, at least at low levels. These infections may alter immune cell function in a way that promotes chlamydial persistence in the host and contributes to the progression of several chronic inflammatory diseases. In this paper, we review the evidence for the growth of Chlamydia in immune cells, particularly monocytes/macrophages and dendritic cells, and describe how infection may affect the function of these cells.

Two-Photon Fluorescence Microscopy Imaging of Cellular Oxidative Stress Using Profluorescent Nitroxides

A range of varying chromophore nitroxide free radicals and their nonradical methoxyamine analogues were synthesized and their linear photophysical properties examined. The presence of the proximate free radical masks the chromophore’s usual fluorescence emission, and these species are described as profluorescent. Two nitroxides incorporating anthracene and fluorescein chromophores (compounds 7 and 19, respectively) exhibited two-photon absorption (2PA) cross sections of approximately 400 G.M. when excited at wavelengths greater than 800 nm. Both of these profluorescent nitroxides demonstrated low cytotoxicity toward Chinese hamster ovary (CHO) cells. Imaging colocalization experiments with the commercially available CellROX Deep Red oxidative stress monitor demonstrated good cellular uptake of the nitroxide probes. Sensitivity of the nitroxide probes to H2O2-induced damage was also demonstrated by both one- and two-photon fluorescence microscopy. These profluorescent nitroxide probes are potentially powerful tools for imaging oxidative stress in biological systems, and they essentially “light up” in the presence of certain species generated from oxidative stress. The high ratio of the fluorescence quantum yield between the profluorescent nitroxide species and their nonradical adducts provides the sensitivity required for measuring a range of cellular redox environments. Furthermore, their reasonable 2PA cross sections provide for the option of using two-photon fluorescence microscopy, which circumvents commonly encountered disadvantages associated with one-photon imaging such as photobleaching and poor tissue penetration.

Cancer risk in persons with HIV/AIDS in India: a review and future directions for research

Background India has a large and evolving HIV epidemic. Little is known about cancer risk in Indian persons with HIV/AIDS (PHA) but risk is thought to be low. Methods To describe the state of knowledge about cancer patterns in Indian PHA, we reviewed reports from the international and Indian literature. Results As elsewhere, non-Hodgkin lymphomas dominate the profile of recognized cancers, with immunoblastic/large cell diffuse lymphoma being the most common type. Hodgkin lymphoma is proportionally increased, perhaps because survival with AIDS is truncated by fatal infections. In contrast, Kaposi sarcoma is rare, in association with an apparently low prevalence of Kaposi sarcoma-associated herpesvirus. If confirmed, the reasons for the low prevalence need to be understood. Cervical, anal, vulva/vaginal and penile cancers all appear to be increased in PHA, based on limited data. The association may be confounded by sexual behaviors that transmit both HIV and human papillomavirus. Head and neck tumor incidence may also be increased, an important concern since these tumors are among the most common in India. Based on limited evidence, the increase is at buccal/palatal sites, which are associated with tobacco and betel nut chewing rather than human papillomavirus. Conclusion With improving care of HIV and better management of infections, especially tuberculosis, the longer survival of PHA in India will likely increase the importance of cancer as a clinical problem in India. With the population's geographic and social diversity, India presents unique research opportunities that can be embedded in programs targeting HIV/AIDS and other public health priorities.