Antibody response to first and second dose of BNT162b2 in a cohort of characterized healthcare workers

BackgroundVaccine-induced population immunity is a key global strategy to control coronavirus disease 2019 (COVID-19). The rapid implementation and availability of several COVID-19 vaccines is now a global health-care priority but more information about humoral responses to single- and double-dose vaccine is needed Methods163 health care workers (HCW) of the Padua University Hospitals, who underwent a complete vaccination campaign with BNT162b2 vaccine were asked to collect serum samples at 12 (t12) and 28 (t28) days after the first inoculum to allow the measurement of SARS-CoV-2 Antibodies (Ab) using chemiluminescent assays against the spike (S) protein and the Receptor Binding Domain (RBD) of the virus, respectively. ResultsSignificant differences were found at t12 for infection-naive and subjects with previous-natural infection who present higher values of specific antibodies, while no significant differences have been found between t12 and t28. No statistically significant difference was found between male and female, while lower Ab levels have been observed in subjects older than 60 years at t12 but not at t28. ConclusionsOur study confirms observed differences in vaccine responses between infection-naive and subjects with previous natural infection at t12 but not for a longer time. The influence of sex and age deserves further studies, even if the relationship with age seems particularly significant.

Analytical and clinical performances of a SARS-CoV-2 S-RBD IgG assay: comparison with neutralization titers

BackgroundSARS-CoV-2 serology presents an important role in understanding the virus epidemiology, in vaccine prioritization strategies and in convalescent plasma therapy. Immunoassays performances have to be accurately evaluated and correlated with neutralizing antibodies to be used as a surrogate measure of neutralizing activity. We investigate the analytical and clinical performance of a SARS-CoV-2 RBD IgG assay, automated on a high throughput platform, and the correlation of the antibodies (Ab) levels with the plaque reduction neutralization (PRNT50) Ab titers. MethodsA series of 546 samples were evaluated by SARS-CoV-2 RBD IgG assay (Snibe diagnostics), including 171 negative and 168 positive SARS-CoV-2 subjects and a further group of 207 subjects of the COVID-19 family clusters follow-up cohort. ResultsAssay precision was acceptable at low and medium levels; linearity was excellent in all the measurement range. Considering specimens collected after 14 days post symptoms onset, overall sensitivity and specificity were 99.0% and 92.5%, respectively. A total of 281 leftover samples results of the PRNT50 test were available. An elevated correlation was obtained between the SARS-CoV-2 RBD IgG assay and the PRNT50 titer at univariate (rho = 0.689) and multivariate (rho = 0.712) analyses. ConclusionsSARS-CoV-2 S-RBD IgG assay achieves elevated analytical and clinical performances, and a strong correlation with sera neutralization activity.

A longitudinal study of healthcare workers' surveillance during the ongoing COVID-19 Epidemics in Italy: is SARS-CoV-2 still a threat for the Health-care System?

ObjectivesIn spring 2020, Northern Italy was the first area outside China to be involved in the SARS-CoV-2 pandemic. This observational study depicts SARS-CoV-2 prevalence and serological curves among first-line healthcare workers (HCWs) at Padua University Hospital (PdUH), North-East Italy. Method344 HCWs, working at the PdUH Emergency Department and Infectious Disease Unit, underwent a SARS-CoV-2 RNA nasopharyngeal swab with paired IgM and IgG antibody detection for 4 consecutive weeks. At every session, a questionnaire recorded symptoms, signs and recent contacts with SARS-CoV-2 patients. Positive cases were followed up for 5 months. ResultsTwenty-seven HCWs (7.84%) had positive serology (Abs) with 12 positive swabs during the study period. Two additional HCWs were positive by swab but without Abs. Fourteen cases (4%) had SARS-CoV-2 infection before the beginning of the study. An HCW with autoimmune disease showed false Ab results. 46% of individuals with Abs reported no symptoms, in accordance with previous population studies. Fever, nasal congestion, diarrhoea and contacts with SARS-CoV-2 individuals correlated to SARS-CoV-2 infection. 96% of Abs+ cases showed persistent positive antibodies 5 months later and none was re-infected. DiscussionCorrect use of PPEs and separate paths for positive/negative patients in the hospital can result in a low percentage of SARS-CoV-2 infections among HCWs, even in high risk settings. Frequent testing for SARS-CoV-2 with nasopharyngeal swabs is worthwhile, irrespective of HCWs symptoms, due to the lack of specificity together with the high percentage of asymptomatic cases. Further studies are needed to elucidate the neutralizing effect of SARS-CoV-2 antibodies.

Estudo Epidemiológico do Câncer Infantojuvenil no Hospital de Câncer de Cascavel Uopeccan entre os Anos 2000 e 2014 / Epidemiological Study of Childhood Cancer in Cascavel Cancer Hospital Uopeccan among 2000 and 2014 / Estudio Epidemiológico del Cáncer Infantil en el Hospital Oncológico de Cascavel Uopeccan entre 2000 y 2014

Introdução: Estudos epidemiológicos de base hospitalar sobre o câncer infantojuvenil são importantes para mostrar o perfil dos pacientes assistidos pelo serviço. Objetivo: Avaliar o perfil clínico-epidemiológico e a sobrevida d e pacientes na faixa etária de 0-19 anos atendidos no Hospital do Câncer de Cascavel Uopeccan (2000-2014). Método: Estudo transversal com avaliação de prontuários para os seguintes desfechos: sexo, idade, cor/raça, outras patologias, histórico familiar de câncer, domicílio, tipo da neoplasia, estadiamento, tratamento, metástases, recidivas, situação do paciente ao final da pesquisa. A estatística descritiva e os testes qui-quadrado e Kaplan-Meier foram aplicados. Resultados: Observou-se maior frequência para meninos (55,2%), faixa etária de 1-4 anos (36,3%), brancos (87%), domicílio urbano (81,6%), leucemia (35,8%) e quimioterapia (50,2%). Ocorreu metástase em 16,41% e recidiva em 22,38%. Não havia relato de histórico familiar de câncer em 47% dos prontuários. Outras patologias foram negadas em 58,9%. Ao final, 55,2% estavam vivos e sem doença. Houve associação estatisticamente significava entre menores de 10 anos com tumores renais e neuroblastoma; maiores de 10 anos com linfomas e neoplasias epiteliais malignas; e entre a situação atual do paciente com metástase, recidivas e estadiamento. Conclusão: Os pacientes analisados na presente pesquisa eram na maioria leucêmicos, do sexo masculino e faixa etária de 1-4 anos. A sobrevida global e a livre de doença foram, respectivamente, de 70,3% e 71,63%

Introduction: Hospital-based epidemiological studies on childhood cancer are important to show the profile of patients cared by the service. Objective: To evaluate the characteristics of cancer patients aged 0-19 years at the Cascavel Cancer Hospital Uopeccan (2000-2014). Method: Cross-sectional study that evaluated medical charts for the following outcomes: gender, age, color/race, comorbidities, family history of cancer, household, cancer type, staging, treatment, metastasis, recurrences, patient's status at the end of study. Descriptive statistics, chi-square and Kaplan-Meier were applied. Results: Boys were more prevalent (55.2%), age range from 1 to 4 years (36.3%), White (87%), urban household (81.6%), with leukemia (35.83%) and in chemotherapy (50.2%). Metastasis occurred in 16.41% and recurrence in 22.38%. There was no report of family history of cancer in 47% of the charts. Other pathologies were denied in 58.9%. In the end, 55.2% were alive and disease-free. There was a statistically significant association between boys younger than 10 years old with renal tumors and neuroblastoma and older than 10 years with lymphomas and malignant epithelial neoplasms and between the current status of the patient with metastasis, relapses, and staging. Conclusion: The patients analyzed in this study were mostly leukemic, males and aged 1-4 years. Global and disease-free survival were, respectively, 70.3% and 71.63%

Introducción: Los estudios epidemiológicos hospitalarios sobre cáncer infantil son importantes para mostrar el perfil de los pacientes atendidos por el servicio. Objetivo: Evaluar las características de los pacientes oncológicos de 0-19 años atendidos en el Hospital do Cáncer de Cascavel Uopeccan (2000-2014). Método: Estudio transversal que evaluó historias clínicas para los siguientes resultados: sexo, edad, color/raza, comorbilidades, antecedentes familiares de cáncer, domicilio, tipo de cáncer, estadificación, tratamiento, metástasis, recurrencias, situación del paciente al final de la investigación. Se aplicó estadística descriptiva, chi-cuadrado y Kaplan-Meier. Resultados: Fueron más prevalentes: niños (55,22%), grupo de edad 1 a 4 años (36,32%), blancos (87,06%), hogares urbanos (81,59%), leucemia (35,83) %) y quimioterapia (50,25%). Hubo metástasis en 16,41% y recidiva en 22,38%. No hubo informes de antecedentes familiares de cáncer en 47% de los pacientes. Se negaron comorbilidades en 58,91%. Al final, 55,23% estaban vivos y sin enfermedad. Hubo asociación estadística entre menores de 10 años con tumores renales y neuroblastoma; mayores 10 años con linfomas y neoplasias epiteliales malignas; e entre la situación del paciente con metástasis, recaídas y estadificación. Conclusión: Los pacientes analizados en esta investigación eran en su mayoría leucémicos, varones y de 1 a 4 años. La supervivencia global y libre de enfermedad fueron, respectivamente, 70,3% y 71,63%

Salivary SARS-CoV-2 antigen rapid detection: a prospective cohort study

BackgroundSARS-CoV-2 quick testing and reporting are now considered relevant for the containment of new pandemic waves. Antigen testing in self-collected saliva might be useful. We compared the diagnostic performance of salivary and naso-pharyngeal swab (NPS) SARS-CoV-2 antigen detection by a rapid chemiluminescent assay (CLEIA) and two different point-of-care (POC) immunochromatographic assays, with that of molecular testing. Methods234 patients were prospectively enrolled. Paired self-collected saliva (Salivette) and NPS were obtained to perform rRT-PCR, chemiluminescent (Lumipulse G) and POC (NPS: Fujirebio and Abbott; saliva: Fujirebio) for SARS-CoV-2 antigen detection. ResultsThe overall agreement between NPS and saliva rRT-PCR was 78.7%, reaching 91.7% at the first week from symptoms onset. SARS-CoV-2 CLEIA antigen was highly accurate in distinguishing between positive and negative NPS (ROC-AUC=0.939, 95%CI:0.903-0.977), with 81.6% sensitivity and 93.8% specificity. This assay on saliva had an overall good accuracy (ROC-AUC=0.805, 95%CI:0.740-0.870), reaching the optimal value within 7 days from symptom onset (Sensitivity: 72%; Specificity: 97%). POC antigen in saliva had a very limited sensitivity (13%), performing better in NPS (Sensitivity: 48% and 66%; Specificity: 100% and 99% for Espline and Abbott respectively), depending on viral loads. ConclusionsSelf-collected saliva is a valid alternative to NPS for SARS-CoV-2 detection not only by molecular, but also by CLEIA antigen testing, for which the highest diagnostic accuracy was achieved in the first week from symptom onset. Saliva is not suitable for POC, although the accuracy of these tests appears satisfactory for NPS with high viral load.

Frequent testing regimen based on salivary samples for an effective COVID-19 containment strategy

Rapid and accurate diagnostic tests are essential for controlling the ongoing COVID-19 pandemic. Although the current gold standard involves testing of nasopharyngeal swabs specimens by nucleic acid amplification test, such as real-time reverse-transcription polymerase chain reaction (rRT-PCR) to detect the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), it presents several limitations that ultimately may translate into a bottleneck in the surveillance regimen. New strategies based on frequent testing using less invasive specimens are urgently needed for containment of the infection. Rapid antigen assay using saliva as a reliable alternative to nasopharyngeal swabs should be proposed as a valuable part of the overall testing strategy.

Potential of vermicompost and limestone in reducing copper toxicity in young grapevines grown in Cu-contaminated vineyard soil.

Foliar fungicide application in grapevines increases the content of heavy metals such as copper (Cu) in vineyard soils, which may reach phytotoxic levels. The application of soil amendments such as limestone and vermicompost may reduce Cu availability and phytotoxicity. The study aimed to assess growth, physiological status and changes in root morphology in young grapevines grown for 12 months in Cu-contaminated soil with and without the application of soil amendments. Samples of a Typic Hapludalf soil were collected in a vineyard with more than 30 years of cultivation. The following treatments were used: 1) control (without amendment), 2) application of organic vermicompost (86.7 g kg-1) and 3) application of limestone (3 Mg ha-1). Grapevines (Paulsen 1103 rootstock) were transplanted and grown for 12 months in PVC soil columns. We assessed parameters of growth, photosynthesis and root morphology. Grapevines grown in soil treated with limestone showed increased growth, dry matter yield and photosynthetic efficiency. The highest Cu concentrations in root tissue were found in grapevines grown in control soil. The application of vermicompost in this study did not alleviate Cu toxicity. Grapevines grown in soil treated with vermicompost showed high manganese (Mn) concentration in shoots before the winter pruning, reflecting the high Mn concentrations in soil solution that caused Mn phytotoxicity, resulting in plant death after the winter pruning. The vermicompost used in this study is not suitable for agronomic use. The use of limestone was an effective strategy to reduce Cu availability and phytotoxicity.

Bond strength to ZLS ceramics at different etching times and cementation protocols after aging / Influência da cimentação e da termociclagem na resistência de união da cerâmica de silicato de lítio reforçada com zircônia

Objective: To assess the influence of 5% hydrofluoric acid etching time (ET), cementation protocol (CP), and thermal cycling (TC) aging on the microshear bond strength (µSBS) of zirconia reinforced lithium silicate ceramic (ZLS) to adhesive resin cement. Material and Methods: Ten VITA Suprinity® ceramic blocks were cut in 120 slices (1.4 mm thickness) and randomly assigned to 12 groups (n = 10) according to the combination of factors (2x3x2 design): etching time (20 or 30 s), cementation protocol (silane + universal adhesive + resin cement; universal adhesive + resin cement; silane + resin cement) and thermal cycling (cycled or no-cycled). RelyX Ceramic Primer and Scotchbond™ Universal Adhesive were used respectively as silane (S) and universal adhesive (Ua). Ceramic surface was etched, and the cementation protocol performed on the delimited bonding area. Then, resin cement (RelyX™ Ultimate Cement [Rc]) cylinders were bonded and light cured. After, specimens were stored in deionized water at 37°C for 7 days and subjected to the µSBS test. Results: Data passed the normality test and three-way ANOVA analysis showed statistical difference (p < 0.01) for isolated; double (ET/TC) (p < 0.05), and triple (p < 0.05) factor interactions. Conclusion: The combination 30s etching-Ua-Rc presented higher adhesive bond strength after thermal aging. (AU)

Objetivo: Avaliar a influência do tempo de condicionamento com ácido fluorídrico 5% (ET), protocolo de cimentação (CP), e envelhecimento por termociclagem (TC) na resistência de união ao microcisalhamento (µSBS) da cerâmica de silicato de lítio reforçada com zircônia (ZLS) ao cimento resinoso adesivo. Material e Método: Dez blocos da cerâmica VITA Suprinity® foram cortados e 120 fatias (com espessura de 1,4 mm) e distribuídos aleatoriamente em 12 grupos (n=10) de acordo com o fator de combinação (2x3x2): tempo de condicionamento (20 ou 30 s), protocolo de cimentação (silano + adesivo universal + cimento resinoso; adesivo universal + cimento resinoso; silano + cimento resinoso) e ciclagem térmica (ciclado ou não ciclado). RelyX Ceramic Primer e adesivo Scotchbond™ Universal foram utilizados respectivamente como silano (S) e adesivo universal (Ua). A superfície cerâmica foi condicionada e o protocolo de cimentação aplicado sobre área de cimentação delimitada. Portanto, cilindros de cimento resinoso (cimento RelyX™ Ultimate [Rc]) foram cimentados e fotopolimerizados. Após, os espécimes foram armazenados em água deionizada a 37°C por 7 dias e submetidos ao teste de µSBS. Resultados: Após teste de normalidade, os dados foram analisados pelo teste ANOVA três critérios mostrando diferença estatística (p < 0.01) para a análise do fator isolado; para a interação entre dois fatores (ET/TC) (p < 0.05), e interação entre os três fatores (p < 0.05). Conclusão: A combinação 30s de condicionamento-Ua-Rc apresentou a maior resistência de união adesiva após a ciclagem térmica.(AU)

Efeito de nanopartículas de trimetafosfato de sódio associadas a dentifrício com concentração reduzida de fluoreto sobre a desmineralização do esmalte bovino: estudos in vitro e in situ / Effect of nano-sized sodium trimetaphosphate associated to low-fluoride dentifrice on bovine enamel demineralization: in vitro and in situ studies

O objetivo deste estudo foi avaliar o efeito de dentifrício com concentração reduzida de fluoreto (250 ppm F) contendo trimetafosfato de sódio nanoparticulado (TMPnano) sobre a desmineralização do esmalte bovino (in vitro e in situ), bem como sua ação no biofilme dentário formado in situ. Para o estudo in vitro, blocos de esmalte bovino (n = 144) foram divididos em 12 grupos de dentifrícios (ph: 7,0): Placebo (sem F ou TMP), 1100 ppm F (1100 - DC - dentifrício convencional), 250 ppm F (250) e, 250 ppm F suplementado com TMP micrométrico (250-TMP) ou TMP nanoparticulado moído por 24h (250-TMPnano24) ou por 48h (250-TMPnano48). O TMP micrométrico e o TMP nanoparticulado foram adicionados à formulação nas concentrações de 0,05%, 0,1% e 0,25%. Os blocos foram submetidos à ciclagem de pH durante cinco dias, sendo o tratamento com suspensões dos respectivos dentifrícios realizados 2x/dia. Para avaliar a perda mineral, a dureza de superfície final (SHf) e a porcentagem de perda de dureza de superfície (%SH) foram determinadas, bem como a concentração de fluoreto (F), cálcio (Ca), fósforo (P) e proporção molar Ca/P no esmalte. Os dados foram submetidos aos testes Kruskal-Wallis e Student-Newman-Keuls (p<0,05). Todas as concentrações de TMPnano reduziram a %SH (p˂0,05). Os dentifrícios 250-TMPnano24 e 250- TMPnano48 levaram a maior retenção de F, Ca, P e Ca/P no esmalte apenas na concentração de 0,05% (p˂0,05); mas, apenas o dentifrício 250-0,05TMPnano48 não apresentou diferença estatística quando comparado ao 1100 para a proporção molar Ca/P no esmalte (p˂0,05). Já para o estudo in situ, 19 voluntários utilizaram aparelhos contendo 4 blocos de esmalte bovino e foram aleatoriamente distribuídos quanto ao dentifrício a ser utilizado: placebo (sem F/TMP), 250 ppm F (250), 250 ppm F suplementado com 0,05% de TMP nanoparticulado triturado por 48h (250-TMPnano) e 1100 ppm F (1100 - DC – dentifrício convencional), sob desafio cariogênico (sacarose 30%, 6x/dia), por 7 dias...

The aim of this study was to evaluate the effect of low-fluoride dentifrices (250 ppm F) containing nano-sized sodium trimetaphosphate (TMPnano) on the demineralization of bovine enamel as well their action on the biofilm formed in situ. For study 1 (in vitro), blocks of bovine enamel (n=144) were randomly divided into 12 experimental groups of dentifrices (pH 7.0). Placebo (no F or TMP), 1,100 ppm F (conventional dentifrice), 250 ppm F (250), 250 ppm F plus micrometric TMP (250-TMP) or nano-sized TMP milled for 24h (250-TMPnano24) or 48h (250-TMPnano48). Micrometric and nano-sized TMP were added to the formulations at concentrations of 0.05%, 0.1% and 0.25%. Blocks were subjected to a pH-cycling regimen and treatments with dentifrice slurries during 7 days. To evaluate mineral loss, surface hardness (SH) and the percentage of surface hardness loss (%SH) were analyzed. Fluoride (F), calcium (Ca), phosphorus (P) and Ca/P molar ratio in enamel were also determined. Data were submitted to Kruskal-Wallis test, followed by StudentNewmanKeuls’ test (p<0.05). All concentrations of TMPnano significantly reduced %SH (p˂0.05). In addition, the dentifrices 250-TMPnano24 and 250-TMPnano48 led to higher F, Ca, P and Ca/P content in enamel only at 0.05% (p˂0.05); but only the 250-0.05TMPnano48 did not show significantly statistical difference when compared with 1,100 in respect of Ca/P molar ratio in enamel (p˂0.05). For study 2 (in situ), nineteen subjects wore palatal appliances containing 4 blocks of bovine enamel, and were randomly assigned to brush their teeth with placebo (no F or TMP), 250 ppm F (250), 250 plus 0.05% nano-sized TMP milled for 48h (250-TMPnano) and 1,100 ppm F (1,100 - conventional dentifrice) dentifrices, under cariogenic challenge (30% sucrose solution, 2 times/day), during 7 days. Post-cariogenic challenge hardness (SHf), the percentage of surface hardness loss (%SH), the integrated loss of subsurface hardness (KHN), as well as fluoride (F)...

Efeito de nanopartículas de trimetafosfato de sódio associadas a dentifrício com concentração reduzida de fluoreto sobre a desmineralização do esmalte bovino: estudos in vitro e in situ / Effect of nano-sized sodium trimetaphosphate associated to low-fluoride dentifrice on bovine enamel demineralization: in vitro and in situ studies

O objetivo deste estudo foi avaliar o efeito de dentifrício com concentração reduzida de fluoreto (250 ppm F) contendo trimetafosfato de sódio nanoparticulado (TMPnano) sobre a desmineralização do esmalte bovino (in vitro e in situ), bem como sua ação no biofilme dentário formado in situ. Para o estudo in vitro, blocos de esmalte bovino (n = 144) foram divididos em 12 grupos de dentifrícios (ph: 7,0): Placebo (sem F ou TMP), 1100 ppm F (1100 - DC - dentifrício convencional), 250 ppm F (250) e, 250 ppm F suplementado com TMP micrométrico (250-TMP) ou TMP nanoparticulado moído por 24h (250-TMPnano24) ou por 48h (250-TMPnano48). O TMP micrométrico e o TMP nanoparticulado foram adicionados à formulação nas concentrações de 0,05%, 0,1% e 0,25%. Os blocos foram submetidos à ciclagem de pH durante cinco dias, sendo o tratamento com suspensões dos respectivos dentifrícios realizados 2x/dia. Para avaliar a perda mineral, a dureza de superfície final (SHf) e a porcentagem de perda de dureza de superfície (%SH) foram determinadas, bem como a concentração de fluoreto (F), cálcio (Ca), fósforo (P) e proporção molar Ca/P no esmalte. Os dados foram submetidos aos testes Kruskal-Wallis e Student-Newman-Keuls (p<0,05). Todas as concentrações de TMPnano reduziram a %SH (p˂0,05). Os dentifrícios 250-TMPnano24 e 250- TMPnano48 levaram a maior retenção de F, Ca, P e Ca/P no esmalte apenas na concentração de 0,05% (p˂0,05); mas, apenas o dentifrício 250-0,05TMPnano48 não apresentou diferença estatística quando comparado ao 1100 para a proporção molar Ca/P no esmalte (p˂0,05). Já para o estudo in situ, 19 voluntários utilizaram aparelhos contendo 4 blocos de esmalte bovino e foram aleatoriamente distribuídos quanto ao dentifrício a ser utilizado: placebo (sem F/TMP), 250 ppm F (250), 250 ppm F suplementado com 0,05% de TMP nanoparticulado triturado por 48h (250-TMPnano) e 1100 ppm F (1100 - DC – dentifrício convencional), sob desafio cariogênico (sacarose 30%, 6x/dia), por 7 dias...

The aim of this study was to evaluate the effect of low-fluoride dentifrices (250 ppm F) containing nano-sized sodium trimetaphosphate (TMPnano) on the demineralization of bovine enamel as well their action on the biofilm formed in situ. For study 1 (in vitro), blocks of bovine enamel (n=144) were randomly divided into 12 experimental groups of dentifrices (pH 7.0). Placebo (no F or TMP), 1,100 ppm F (conventional dentifrice), 250 ppm F (250), 250 ppm F plus micrometric TMP (250-TMP) or nano-sized TMP milled for 24h (250-TMPnano24) or 48h (250-TMPnano48). Micrometric and nano-sized TMP were added to the formulations at concentrations of 0.05%, 0.1% and 0.25%. Blocks were subjected to a pH-cycling regimen and treatments with dentifrice slurries during 7 days. To evaluate mineral loss, surface hardness (SH) and the percentage of surface hardness loss (%SH) were analyzed. Fluoride (F), calcium (Ca), phosphorus (P) and Ca/P molar ratio in enamel were also determined. Data were submitted to Kruskal-Wallis test, followed by StudentNewmanKeuls’ test (p<0.05). All concentrations of TMPnano significantly reduced %SH (p˂0.05). In addition, the dentifrices 250-TMPnano24 and 250-TMPnano48 led to higher F, Ca, P and Ca/P content in enamel only at 0.05% (p˂0.05); but only the 250-0.05TMPnano48 did not show significantly statistical difference when compared with 1,100 in respect of Ca/P molar ratio in enamel (p˂0.05). For study 2 (in situ), nineteen subjects wore palatal appliances containing 4 blocks of bovine enamel, and were randomly assigned to brush their teeth with placebo (no F or TMP), 250 ppm F (250), 250 plus 0.05% nano-sized TMP milled for 48h (250-TMPnano) and 1,100 ppm F (1,100 - conventional dentifrice) dentifrices, under cariogenic challenge (30% sucrose solution, 2 times/day), during 7 days. Post-cariogenic challenge hardness (SHf), the percentage of surface hardness loss (%SH), the integrated loss of subsurface hardness (KHN), as well as fluoride (F)...