Benzidine staining: method to detect neutrophils in whole saliva of patients with renal disease

Benzidine staining allows for the quantification of peroxidise positive polymorphonuclear leukocytes in whole saliva from patients with renal disease. This is a practical tool for monitoring oral health in this high risk population.Fil: Cardoso, Estela M. del Luján. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina. Universidad de Buenos Aires. Facultad de Odontología; ArgentinaFil: Arregger, Alejandro Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Budd, D.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Elverdín, Juan Carlos. Universidad de Buenos Aires. Facultad de Odontología; ArgentinaFil: Tumilasci, G.. Universidad de Buenos Aires. Facultad de Odontología; ArgentinaFil: Contreras, Liliana Noemí. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina. Universidad de Buenos Aires. Facultad de Odontología; Argentin

Debilitating consequences of drooling

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Standardization of a simple method to study whole saliva: clinical use in different pathologies.

En este trabajo se describe la normatización de un método para determinar flujo salival en humanos utilizando saliva total obtenida del piso de la boca mediante un eyector dental descartable y una bomba de vacío (equipo dental). En este estudio se evaluaron 40 sujetos sanos de ambos sexos y 51 pacientes con diversas patologías (Síndrome de Sjögren, Disfunción Tiroidea, Diabetes Mellitus). Se demostró que el flujo salival basal era estable a partir de los primeros 5 minutos de colocado el eyector en la cavidad bucal. No se encontraron diferencias significativas en el flujo salival basal comparando los sexos, siendo independiente de la intensidad del vacío efectuado por la bomba. El flujo de saliva total estimulada fue determinado durante 3 minutos, luego de los primeros 5 minutos de colocado el eyector en la boca. El estímulo se efectuó adosando en la cara dorsal de la lengua discos de papel absorbente, embebidos en ácido cítrico al 2 %. El uso de este método en pacientes con Síndrome de Sjögren confirmó la reducción del flujo salival respecto a los sujetos sanos. Los pacientes hipotiroideos y con neuropatía diabética demostraron disminución del flujo salival.The present study describes a methodology to assess the salivary flow rate in humans. Whole saliva was obtained from the floor of the mouth with a plastic dental ejector and a vacuum pump. Forty healthy subjects of both sexes and 51 patients with different pathologies (Sjögren Syndrome, Thyroid Dysfunction, Diabetes Mellitus) were included in the study. It was demonstrated that basal salivary flow rate was stable five minutes after the insertion of the oral ejector. Salivary flow rate did not show significant differences between sexes and was independent of the negative pressure level of the vacuum pump. Stimulated salivary flow rate was quantified over a period of 3 minutes, starting 5 minutes after the introduction of the oral device. The stimulus was paper filter disks soaked in citric acid (2%) placed on the tongue dorsum. The use of this method confirmed the reduction of salivary flow rate in patients with Sjögren Syndrome. In addition, a significant reduction in salivary flow rate was observed in patients with primary thyroid insufficiency and peripheral neuropathy secondary to Diabetes Mellitus.Fil: Tumilasci, Omar Rene. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Fisiología; ArgentinaFil: Cardoso, Estela M. del Luján. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Contreras, Liliana Noemí. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Belforte, Juan Emilio. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Fisiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; ArgentinaFil: Arregger, Alejandro Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Ostuni, Mariano A.. Inserm; Franci

Diurnal and nocturnal drooling in Parkinson’s disease

Drooling as symptom of Parkinson’s disease (PD) has thus far been poorly defined. This uncertainty is reflected by high variations in published prevalence rates. The aim of this study was to investigate the prevalence of saliva loss versus accumulation of saliva as a possible preliminary stage, and diurnal drooling versus nocturnal drooling. In addition, we evaluated the association between drooling severity and the severity of facial and oral motor disorders. We collected age, disease duration, UPDRS III and Hoehn & Yahr stage from 104 consecutive outpatients with PD. Diurnal and nocturnal drooling was evaluated with a validated questionnaire (ROMP-saliva). A speech pathologist, blinded for drooling severity, rated facial expression, involuntary mouth opening and difficulty with nose breathing and also interviewed patients about sleeping position and nose-breathing during the night. Thirty patients (29%) had no complaints with saliva control (‘non-droolers’), 45 patients (43%) experienced accumulation of saliva or only nocturnal drooling (‘pre-droolers’), and 29 (28%) had diurnal drooling (24 of which also drooled during the night; ‘droolers’). The droolers had longer disease duration (10 vs. 7 years, p = 0.01) and drooling was independently associated with involuntary mouth opening (OR = 2.0; 95% CI 1.02–3.99) and swallowing complaints (OR = 1.2; 95% CI 1.03–1.31). Diurnal drooling—defined as dribbling of saliva while awake—is present in about 28% of PD patients. This is less than usually reported. Diurnal drooling typically appeared later in the disease course. The association with oral motor behavior should encourage the development of behavioral treatment approaches

Improving VSV virotherapy in chronic lymphocytic leukemia with small-molecule BCL-2 inhibitors

Oncolytic virus therapy is a new form of cancer treatment that uses viruses that preferentially infect and lyse cancer cells. Vesicular stomatitis virus (VSV) is a strong oncolytic virus candidate that infects multiple tumor cells, produces rapid viral replication in malignant cells and spreads quickly in the tumor. Defects in the interferon (IFN) antiviral pathway are common in tumor cells and such defects are accountable for the sensitivity to VSV infection and replication in several malignant cells. The intrinsic mitochondrial apoptotic pathway plays a crucial role in VSV-induced apoptosis and disturbance of this pathway is responsible for resistance to VSV oncolysis of cancer cells. The antiapoptotic protein B-cell lymphoma 2 (BCL-2) is commonly overexpressed in tumor cells, especially in hematological malignancies, and is strongly related to resistance to cancer therapy. Chronic lymphocytic leukemia (CLL) is an accumulative disease of mature-looking CD5+/CD19+ lymphocytes, caused by defects in apoptosis rather than increase in proliferation. CLL patients express high levels of the BCL-2 protein which correlates with poor treatment outcome. Small-molecule BCL-2 inhibitors showed promising anticancer properties in preclinical models. A phase I clinical trial demonstrated modest activity against CLL. Based on our observation that CLL cells are resistant to VSV-induced cell death due to overexpression of BCL-2, we hypothesized that inhibition of BCL-2 could restore VSV oncolytic potential in primary CLL cells. In fact, BCL-2 inhibitors EM20-25IVand obatoclax sensitized primary CLL cells to VSV-induced cell death. Mechanistically, while VSV infection triggered Phorbol-12-myristate-13-acetate-induced protein 1 (NOXA) upregulation, obatoclax blocked the ability of BCL-2 to dimerize with the proapoptotic BCL2-associated X protein (BAX). Together, NOXA expression and BAX release were able to efficiently induce apoptosis. Moreover, our data demonstrated a direct interaction between NOXA and BAX. Together, these data indicate that the use of BCL-2 inhibitors may improve VSV oncolysis in apoptosis-resistant hematological malignancies characterized by overexpression of anti-apoptotic proteins such as BCL-2.Oncolytic virus therapy is a new form of cancer treatment that uses viruses that preferentially infect and lyse cancer cells. Vesicular stomatitis virus (VSV) is a strong oncolytic virus candidate that infects multiple tumor cells, produces rapid viral replication in malignant cells and spreads quickly in the tumor. Defects in the interferon (IFN) antiviral pathway are common in tumor cells and such defects are accountable for the sensitivity to VSV infection and replication in several malignant cells. The intrinsic mitochondrial apoptotic pathway plays a crucial role in VSV-induced apoptosis and disturbance of this pathway is responsible for resistance to VSV oncolysis of cancer cells. The antiapoptotic protein B-cell lymphoma 2 (BCL-2) is commonly overexpressed in tumor cells, especially in hematological malignancies, and is strongly related to resistance to cancer therapy. Chronic lymphocytic leukemia (CLL) is an accumulative disease of mature-looking CD5+/CD19+ lymphocytes, caused by defects in apoptosis rather than increase in proliferation. CLL patients express high levels of the BCL-2 protein which correlates with poor treatment outcome. Small-molecule BCL-2 inhibitors showed promising anticancer properties in preclinical models. A phase I clinical trial demonstrated modest activity against CLL. Based on our observation that CLL cells are resistant to VSV-induced cell death due to overexpression of BCL-2, we hypothesized that inhibition of BCL-2 could restore VSV oncolytic potential in primary CLL cells. In fact, BCL-2 inhibitors EM20-25IVand obatoclax sensitized primary CLL cells to VSV-induced cell death. Mechanistically, while VSV infection triggered Phorbol-12-myristate-13-acetate-induced protein 1 (NOXA) upregulation, obatoclax blocked the ability of BCL-2 to dimerize with the proapoptotic BCL2-associated X protein (BAX). Together, NOXA expression and BAX release were able to efficiently induce apoptosis. Moreover, our data demonstrated a direct interaction between NOXA and BAX. Together, these data indicate that the use of BCL-2 inhibitors may improve VSV oncolysis in apoptosis-resistant hematological malignancies characterized by overexpression of anti-apoptotic proteins such as BCL-2

Detection of α-Synuclein in saliva: the Importance of preanalytical assessment

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