DISTINCT CELLULAR MIGRATION INDUCED BY Leishmania infantum chagasi AND SALIVA FROM Lutzomyia longipalpis IN A HEMORRHAGIC POOL MODEL

Recruitment of a specific cell population after Leishmania infection can influence the outcome of the disease. Cellular migration in response to Leishmania or vector saliva has been reported in air pouch model, however, cellular migration induced by Leishmania associated with host's blood and vector saliva in this model has not been described. Herein we investigated cellular migration into air pouch of hamster after stimulation with combination of L. chagasi and host's blood and Lutzomyia longipalpis saliva. Migration induced by saliva was 3-fold more than those induced by L. chagasi alone. Additionally, L. chagasi associated with blood and saliva induced significantly even more leukocytes into air pouch than Leishmania alone. L. chagasi recruited a diverse cell population; however, most of these cells seem to have not migrated to the inflammatory exudate, remaining in the pouch lining tissue. These results indicate that L. chagasi can reduce leukocyte accumulation to the initial site of infection, and when associated with vector saliva in the presence of blood components, increase the influx of more neutrophils than macrophages, suggesting that the parasite has developed a strategy to minimize the initial inflammatory response, allowing an unlimited progression within the host. This work reinforces the importance of studies on the salivary components of sand fly vectors of leishmaniasis in the transmission process and the establishment of the infection.

Cortisol and Anxiety Response to a Relaxing Intervention on Pregnant Women Awaiting Amniocentesis

Background: Stress and anxiety during pregnancy have been associated with premature and low birth weight babies, presumably through fetus over exposion to glucocorticoids. Antenatal stress also seems to have long-term effects upon infant development and adult health. However, medication for stress may carry risks to the expectant mother, therefore the efficacy of nonpharmacological interventions should be investigated. Methods: Pregnant women (n = 154) awaiting amniocentesis, were randomly assigned in the morning and the afternoon to three groups for 30 min: (1) listening to relaxing music, (2) sitting and reading magazines, and (3) sitting in the waiting-room. Before and after that period, they completed the Spielberger’s State and Trait anxiety inventory and provided blood samples for cortisol. The groups were then compared regarding change in cortisol levels and anxiety. Results: Maternal cortisol and state anxiety were correlated (r = 0.25, p = 0.04) in the afternoon, but not in the morning. The larger decreases in cortisol occurred in the music group ( 61.8 nmol/L, ANOVA: p = 0.01), followed by magazine, being differences among groups more pronounced in the morning. Women in the music group also exhibited the greater decreases in state anxiety ( p < 0.001). Younger mothers with less gestational age were on average the most anxious, and also the ones with greater decreases in cortisol and anxiety levels after relaxation. Conclusion: A relaxing intervention as short as 30 min, especially listening to music, decreases plasma cortisol and self-reported state anxiety score. Pregnant women might benefit from the routine practice of relaxation in the imminence of clinical stressful events.

LACK OF ASSOCIATION BETWEEN HERPESVIRUS DETECTION IN SALIVA AND GINGIVITIS IN HIV‑INFECTED CHILDREN

The aims of this study were to compare the detection of human herpesviruses (HHVs) in the saliva of HIV-infected and healthy control children, and to evaluate associations between viral infection and gingivitis and immunodeficiency. Saliva samples were collected from 48 HIV-infected and 48 healthy control children. Clinical and laboratory data were collected during dental visits and from medical records. A trained dentist determined gingival indices and extension of gingivitis. Saliva samples were tested for herpes simplex virus types 1 and 2 (HSV-1 and HSV-2), varicella zoster virus (VZV), Epstein-Barr virus (EBV), and cytomegalovirus (CMV) by nested polymerase chain reaction assays. Thirty-five HIV-infected and 16 control children had gingivitis. Seventeen (35.4%) HIV-infected children and 13 (27%) control children were positive for HHVs. CMV was the most commonly detected HHV in both groups (HIV-infected, 25%; control, 12.5%), followed by HSV-1 (6.2% in both groups) and HSV-2 (HIV-infected, 4.2%; control, 8.3%). The presence of HHVs in saliva was not associated with the presence of gingivitis in HIV-1-infected children (p = 0.104) or healthy control children (p = 0.251), or with immunosuppression in HIV-infected individuals (p = 0.447). Gingivitis was correlated with HIV infection (p = 0.0001). These results suggest that asymptomatic salivary detection of HHVs is common in HIV-infected and healthy children, and that it is not associated with gingivitis.

Observações complementares sôbre a presença do Toxoplasma gondii na saliva de pacientes com toxoplasmose

Através de inoculação no peritônio de camundongos, demonstraram, os autores a presença do Toxoplasma gondii na saliva de 12 (60%) indivíduos com toxoplasmose, em estudo no qual 20 doentes foram considerados. Esta verificação é comunicada como complementação a outra, anterior e preliminar, relativa ao mesmo assunto. A importância da presença do parasito na saliva mereceu destaque, em virtude do significado que ela pode ter em relação à transmissão da toxoplasmose.

Cortisol e testosterona no plasma da grávida e no líquido amniótico: variação com o relaxamento e correlação com o rácio digital 2D:4D no recém-nascido

RESUMO: Conclusão - Este estudo sugere que mulheres grávidas beneficiam da prática rotineira de 30 min de relaxamento perante a eminência de um acontecimento indutor de stress. O benefício será tanto psicológico como fisiológico e será mais marcado de manhã do que de tarde. As mulheres com maior tendência ansiosa, tipicamente as mais jovens e com menos idade gestacional, serão as que mais beneficiam. Entre os tipos de intervenção experimentados, todos eles não dispendiosos, a audição de música foi globalmente o mais eficaz para o propósito pretendido. A variação do cortisol e a variação da testosterona estão altamente correlacionadas, e este efeito poderá ser explorado em futuras intervenções relaxantes. Programas curtos de relaxamento poderão conduzir a benefícios para o bem-estar psicológico e para a saúde da grávida, com benefícios adicionais no desfecho da gravidez. O rácio digital 2D:4D do recém-nascido apresenta características sexualmente dimórficas, sendo mais baixo em rapazes que em raparigas, confirmando a influência dos androgénios pré-natais, mas há grande sobreposição entre os sexos pelo que, isoladamente, não pode ser utilizado como marcador de risco. No entanto, se associado a outros marcadores biológicos, como o peso ao nascer e a circunferência cerebral, entre outros, poderá ser um parâmetro a valorizar desde cedo, possivelmente no acompanhamento pediátrico. Nas raparigas, o RD da mãe e a testosterona no LA explicam uma proporção significativa do seu 2D:4D. Em ambos os sexos, o rácio 2D:4D aumenta após o nascimento e o dimorfismo sexual observado com base em amostras de crianças com mais de dois anos de idade, é afectado pela testosterona pós-natal, especialmente no que se refere à mão direita.----------ABSTRACT: Conclusions - This study suggests that pregnant women would benefit from the routine practice of 30min relaxation when at the imminence of a stressful event. The benefit would be both psychological and physiological and would be more pronounced in the morning than in the afternoon. Women tending to be more anxious, typically younger and at a lower gestational age, are the ones that benefit the most. Among the interventions investigated, all being non-expensive and easy to implement, listening to relaxing music was globally the most efficient for the benefit intended. The variation of cortisol and the variation of testosterone are significantly correlated, an association that might be used in future relaxation interventions. It is feasible to conceive short relaxation programs aimed at benefiting psychological well-being and health in pregnancy, with additional benefits for pregnancy outcome. The digital ratio 2D:4D is sexually dimorphic, being lower in males than in females, confirming the influence of pre-natal androgens. There is however large overlap between sexes, rendering the 2D:4D a risk marker not to be used in isolation but rather in association with other markers, like for instance the weight at birth and the head circumference, wherein it may be a relevant parameter for pediatric follow-up. In newborn females, a significant amount of variability in 2D:4D is accounted for by their mother’s digital ratio and AF testosterone. In both sexes, the 2D:4D ratio increases following birth and sexual dimorphism,as assessed from samples of subjects with more than 2 years of age, is affected by post-natal testosterone, especially in what concerns the right hand.

Diagnosis of rubella infection by detecting specific immunoglobulin M antibodies in saliva samples: a clinic-based study in Niterói, RJ, Brazil

This study was designed to investigate whether saliva could be a feasible alternative to serum for the diagnosis of recent rubella infection in a clinic setting. Forty-five paired blood and saliva samples collected 1 to 29 days after onset of illness were tested for specific immunoglobulin (Ig) M by antibody-capture radioimmunoassay (MACRIA). Rubella IgM was detected in all serum samples and in 38 (84.4%) saliva specimens. Forty-six serum and saliva samples from other patients with rash diseases were tested by MACRIA for control purposes and two saliva specimens were reactive. The saliva test had specificity of 96%. These results indicate that salivary IgM detection may be a convenient non-invasive alternative to serum for investigation of recent rubella cases, especially for disease surveillance and control programmes.

Nitric oxide levels are not changed in saliva of patients infected with hepatitis C virus

The aim of this investigation was to determine nitric oxide metabolite levels in saliva samples from hepatitis C virus-positive patients in an attempt to test the hypothesis if increased levels of nitric oxide metabolites correlates with the presence of HCV-RNA in saliva. Saliva of 39 HCV-positive patients and 13 HCV-negative patients, without clinical or laboratorial evidence of liver disease were tested for nitric oxide metabolites. HCV-RNA was detected in serum and saliva by a RT-PCR method and nitric oxide level was determined by evaluation of its stable degradation products, nitrate and nitrite. No differences were found between the concentration of nitrite in saliva from HCV patients and controls, in despite of the presence or not of HCV RNA in saliva. Patients with HCV and cirrhosis had higher concentrations of nitrite but not significantly different from the control group or the groups of anti-HCV patients without cirrhosis. Increased levels of nitrite were not detected in anti-HCV positive patients, an indirect indication that chronic sialoadenitis are infrequent in these patients or occurs with low intensity not sufficient to increase nitric oxide metabolite levels in saliva.

Detecção do gene mecA em estafilococos coagulase negativa resistentes à oxacilina isolados da saliva de profissionais da enfermagem

Estafilococos coagulase negativa estão frequentemente associados às infecções nosocomiais e os profissionais da saúde podem ser reservatório e dissemina-los no hospital e comunidade. O objetivo desse estudo foi identificar espécies de estafilococos coagulase negativa isolados da saliva de profissionais da enfermagem, determinar o perfil de resistência e detectar o gene mecA. Foram selecionados 100 estafilococos coagulase negativa, sendo 41 identificados como Staphylococcus epidermidis, 25 Staphylococcus saprophyticus, 18 Staphylococcus haemolyticus, 8 Staphylococcus cohnii, 4 Staphylococcus lugdunenses, 3 Staphylococcus capitis, e 1 Staphylococcus Simulans. Desses, 32% apresentaram resistência à oxacilina, 84,4% à mupirocina, 32% à cefoxitina, e todos sensíveis a vancomicina. Dos estafilococos coagulase negativa resistentes à oxacilina, 93,7% desenvolveram-se no agar oxacilina (6µg/ml) e o gene mecA foi detectado em 75%. Os resultados sinalizam que maiores investimentos devem ser direcionados a identificação das espécies de estafilococos coagulase negativa nas instituições de saúde e na comunidade.

PCR detection of multiple human herpesvirus DNA in saliva from HIV-infected individuals in Teresina, State of Piauí, Brazil

INTRODUCTION: Human herpesviruses are frequently associated with orofacial diseases in humans (HSV-1, EBV, CMV and HHV-8), some can also cause systemic disease (CMV and HHV-8). The transmission of these viruses occurs by contact with infected secretions, especially saliva. Human immunodeficiency virus infection is associated with an increased risk of HHVs and related diseases. METHODS: This work aimed to detect HSV-1, EBV, CMV and HHV-8 DNA in saliva of HIV-infected patients from Teresina, northeast Brazil, by PCR and compare these findings with age and sex matched HIV-seronegative individuals. RESULTS: No difference in prevalence was verified between HHV detection in the saliva of HIV-seropositive individuals and controls. The individual frequencies of these viruses in these two populations were different. HIV seropositivity correlated positively with the presence of CMV (OR: 18.2, p= 0.00032) and EBV (OR: 3.44, p= 0.0081). No association between CD4 counts and the prevalence of HHVs in the saliva was observed; however, a strong association was determined between seropositivity and the presence of multiple HHV DNAs in saliva (OR: 4.83, p = 0.0028). CONCLUSIONS: These findings suggest the asymptomatic salivary shedding of HHVs is a common event between HIV-seropositive and seronegative individuals from Teresina, Piauí, Brazil, and, especially for HIV-seropositive patients, saliva is a risk factor for the acquisition/transmission of multiple HHVs.

The use of saliva as a practical and feasible alternative to urine in large-scale screening for congenital cytomegalovirus infection increasesinclusion and detection rates

INTRODUCTION: Although urine is considered the gold-standard material for the detection of congenital cytomegalovirus (CMV) infection, it can be difficult to obtain in newborns. The aim of this study was to compare the efficiency of detection of congenital CMV infection in saliva and urine samples. METHODS: One thousand newborns were included in the study. Congenital cytomegalovirus deoxyribonucleic acid (DNA) was detected by polymerase chain reaction (PCR). RESULTS: Saliva samples were obtained from all the newborns, whereas urine collection was successful in only 333 cases. There was no statistically significant difference between the use of saliva alone or saliva and urine collected simultaneously for the detection of CMV infection. CONCLUSIONS: Saliva samples can be used in large-scale neonatal screening for CMV infection.

Aplicação do Seis Sigma na avaliação da inexatidão dos resultados laboratoriais do parâmetro cortisol sérico

No desenvolvimento deste caso de estudo, dentro do Programa Nacional de Avaliação Externa da Qualidade do Instituto Nacional de Saúde Doutor Ricardo Jorge utilizou-se o Seis Sigma como metodologia e como métrica, de maneira a estruturar de forma intuitiva a sua aplicação na resolução de problemas e definir o principal objetivo. Como metodologia, o Seis Sigma recorre a diversas técnicas e ferramentas da qualidade, que visam a melhoria contínua de procedimentos. Enquanto métrica, o Seis Sigma apresenta uma meta a atingir para a qualidade. O ideal seria atingir-se a meta dos Seis Sigma, que significa 3,4 defeitos por milhão de oportunidades (taxa de defeitos de um determinado procedimento). Para reduzir a variabilidade de resultados entre laboratórios, referentes ao parâmetro Cortisol Sérico, aplicou-se o ciclo DMAIC (Define – Measure – Analyze – Improve – Control), a que correspondem cinco etapas diferentes. Em cada uma destas etapas utilizam-se as técnicas e ferramentas da qualidade, como referido anteriormente acerca do Seis Sigma enquanto metodologia, com o objetivo de melhorar processos. Inicia-se a primeira etapa com a identificação do problema e finaliza-se a última com o controlo e monitorização dos efeitos provocados pelas ações de melhoria implementadas. O objetivo dos programas de Avaliação Externa da Qualidade do Programa Nacional de Avaliação Externa da Qualidade é avaliar a performance quanto à variabilidade/uniformidade de resultados entre os laboratórios participantes. É de extrema importância a harmonização de resultados clínicos entre laboratórios, pois são estes resultados que fornecem informações ao profissional de saúde, sobre o estado de saúde do paciente, auxiliando no diagnóstico, tratamento e controlo de doenças. Caso isto não aconteça, será um fator crítico que pode fazer a diferença na saúde pública. Foram identificadas as causas de erros sistemáticos, causas de inexatidão laboratorial, a nível de métodos, equipamentos, reagentes e calibradores utilizados em laboratório e implementadas ações de melhoria mais relevantes. No final obteve-se um nível de qualidade sigma superior ao inicial, tal como tinha sido previsto nos objetivos. O nível da qualidade deve ser controlado ao longo do tempo, com o objetivo de identificar novas anomalias e melhorar continuamente.

Cortisol, family coping and burden in informal caregivers of addicts

This study focuses on the prospective mediation role of family coping between burden and cortisol levels in informal caregivers of addicts as well as on the feasible use of two different ways to analyse the salivary cortisol levels. Participants were 120 Portuguese informal caregivers of addicts. The cortisol samples were collected at awakening, 45 minutes later and after a 30 minute presentation of images taken from the International Affective Picture System. Family coping and caregiver burden were measured using the Portuguese versions of the Caregiver Reaction Assessment, and the Family Crisis Oriented Personal Evaluation Scale. Cortisol samples were collected in salivettes and the results were computed in order to determine the Area Under the Curve scores (AUCg, AUCi). Results found family coping to be negatively correlated with burden and AUCg levels (i.e. overall intensity) and positively correlated with either AUCg and AUCi (i.e. change over time). The mediation model revealed that family coping was a partial mediator in the relationship between the burden and AUCg levels. Therefore, Family Coping appears to be an essential variable in understanding the stress response and should be considered in further studies and interventions. In addition, the use of two different formulas for calculating cortisol levels provided important new information concerning the relationship between cortisol, burden and family coping. It seems that burden has a more profound effect on the overall intensity of the neuroendocrine response to caregiver stress and not so much on the sensitivity of the system.

Influence of saliva and mucin on the adhesion of Candida oral clinical isolates

Objectives: This research work intends to clarify the role of artificial saliva, in particularly the role of mucin, a salivary protein, on the surface properties and adhesion ability of Candida spp. oral clinical isolates to abiotic surfaces. Methods: Four oral clinical isolates of Candida spp. were used: two Candida albicans strains (AC; AM) and two Candida parapsilosis strains (AD; AM2). The strains were isolated from patients using oral prosthesis. The microorganisms were cultured in the absence or presence of mucin and artificial saliva, and their adhesion to an abiotic surface (coated with mucin and artificial saliva) was evaluated. Results: The presence of mucin per se onto the abiotic surface decreased the adhesion of all strains, although the combination of mucin with artificial saliva had reduced this effect. No direct correlation between adhesion and the surface free energies of adhesion of the microorganisms was found. Significance: Candida spp. were human commensal microorganisms that became pathogenic when the host immune defenses were compromised. Medical devices were colonized by Candida spp. particularly, oral prostheses, which might lead to the degradation of the prostheses and systemic infections. The salivary secretions that constantly cover the oral cavity influenced Candida spp. adhesion process. Therefore, it was important to understand the interactions between Candida spp., salivary proteins and the characteristic of oral prosthesis when developing materials for oral prostheses.

PDMS biofunctionalization study for the development of a microfluidic device: Application to salivary cortisol

The authors acknowledge to Sofia Neves from ICVS for her help in the antibodies selection.