Comparative study of oral and salivary parameters in patients with and without loss of bone mass

Abstract Osteoporosis is an insidious and increasingly prevalent disease that can cause fractures and affect patients' quality of life. The current study comparatively evaluates patients with and without loss of bone mass in terms of salivary calcium, viscosity, and pH. A controlled cross-sectional study was conducted in two groups of 32 postmenopausal women subjected to a bone densitometry scan and later referred for dental management at the Federal University of Minas Gerais, Brazil. The patients were assigned to two groups: Group 1 - patients with low bone mineral density (BMD) and Group 2 - patients without bone mineral changes. The following salivary parameters were evaluated: calcium concentration, flow rate, viscosity, pH, and average total protein. An oral examination was performed for assessment of DMFT variables and tongue coating. Data were analyzed using descriptive and inferential statistics, adopting a p-value < 0.05. The patients' mean age was 60 years (± 7.35). Salivary flow, pH, and viscosity were similar among the groups. Average total protein was 14.8 mg/mL and 19.0 mg/mL in Groups 1 and 2, respectively. Tongue coating and salivary calcium levels were significantly higher in Group 1 (p < 0.001). Salivary calcium is an important screening tool and may eventually be used for the diagnosis of bone mineral changes.

Marcadores e mecanismos de resistência à terapia neoadjuvante e o papel das vesículas extracelulares secretadas pelas células tumorais no câncer de reto / Biomarkers and mechanisms of resistance to neoadjuvnat therapy and the role of extracelular vesicles secreted by tumor cells in rectal cancer

O câncer de reto é o segundo tumor mais comum no intestino grosso correspondendo a um terço do total de casos de câncer colorretal (CCR). Pacientes com câncer de reto em estádios II e III são tratados com radioquimioterapia neoadjuvante seguida de ressecção cirúrgica do tumor. Análises das peças cirúrgicas ressecadas mostraram que apenas 10-45% dos pacientes obtém resposta patológica completa (RCp) à terapia neoadjuvante, estando essa associada com uma diminuição da recorrência local, melhora da sobrevida livre de doença e aumento na preservação esfincteriana. Apesar da melhora na sobrevida nas últimas décadas, a resposta à terapia neoadjuvante continua variável e imprevisível e não é possível identificar e separar clinicamente os grupos de pacientes que terão ou não resposta completa ao tratamento neoadjuvante. Além disso, os mecanismos de resistência à radioquimioterapia nos tumores de reto são pouco compreendidos. Dessa forma, o objetivo principal deste estudo foi identificar marcadores e mecanismos celulares relacionados à resistência à terapia neoadjuvante em adenocarcinoma de reto e o papel das vesículas extracelulares (VEs) nesse processo. O estudo proteômico comparativo entre biópsias obtidas de tumores pré-tratamento com o tumor residual removido cirurgicamente pós-tratamento radioquimioterápico mostrou uma importante alteração no perfil de expressão proteica. Entre as proteínas que aumentam a expressão após a neoadjuvância estão as proteínas de reparo de dano de DNA, Ku70 e Ku80, e a proteína de tráfego intracelular Rab5C. Em um modelo in vitro, foi demonstrado que Rab5C orquestra um mecanismo de resistência à radioterapia nos tumores de reto através da modulação da internalização de EGFR promovida por radiação ionizante (RI). O EGRF intracelular por sua vez é essencial para regular a expressão de Ku70 e Ku80 e a resistência celular à RI. Estes dados apontam Rab5C e EGFR como potenciais alvos terapêuticos para sensibilizar células de câncer de reto resistentes ao tratamento neoadjuvante. Também foi observado que a RI promove alterações epigenéticas predominantemente de hipometilação, e entre os genes alterados estão SPG20 e TBC1D16, sendo o primeiro importante para a internalização de EGFR e o segundo para a regulação de Rab5C e modulação de EGFR. O perfil de expressão proteica foi ainda comparado entre biópsias pré-tratamento de pacientes com RCp e sem resposta patológica, e o resultado mostrou que esses dois grupos de pacientes apresentam um diferente perfil de expressão proteica. Nos pacientes com RCp as proteínas com aumento da expressão estão atuando em vias que favorecem a resposta à terapia, como a detoxificação de glutationa e degradação de glicogênio, enquanto as proteínas com aumento da expressão em pacientes sem RCp estão envolvidas em vias do metabolismo energético do tumor as quais contribuem para a resistência tumoral à terapia. As diferenças observadas nestes grupos devem ser amplamente exploradas uma vez que podem ser marcadores preditivos de resposta ao tratamento radioquimioterápico. A realização de estudos funcionais foi viabilizada pela geração de um modelo celular de tumor de reto resistente à radioterapia. Ao analisar as VEs secretadas por estas células foi observado que a RI não altera a quantidade e o tamanho médio das VEs secretadas, porém é capaz de alterar o carregamento proteico das mesmas. De fato, as VEs de células irradiadas apresentam um perfil proteico diferente quando comparadas as VEs de células não irradiadas, onde encontramos aumento da expressão de Ku70, Ku80 e Rab5C, além das metiltransferases NSUN2 e GLYM nas VEs de células pós RI. Interessantemente, as VEs secretadas por células irradiadas são capazes de transmitir a resistência à RI às células não irradiadas. Além disso os resultados mostraram que o tratamento com VEs de células irradiadas promove metilação em 98% do DNA avaliado em células SW837 em comparação ao tratamento com VEs de células não irradiadas. Os genes hipermetilados estão envolvidos em vias relacionadas ao sistema imune, como a apresentação de antígeno, sinalização de imunodeficiência primária e maturação de células dendríticas. Por fim, foi identificado que a expressão da proteína A33 está relacionada ao grau de diferenciação dos tumores colorretais, e que essa proteína está presente em VEs secretadas por células de adenocarcinoma de reto, indicando que a mesma pode ser usada para isolar VEs específicas do tecido colorretal. Os dados obtidos neste trabalho apontam mecanismos relacionados à resistência à terapia neoadjuvante no adenocarcinoma de reto e que em conjunto permitirão identificar novos alvos terapêuticos com potencial de melhorar a resposta à radioquimioterapia, além de identificar marcadores de resposta à terapia neoadjuvante antes do tratamento e dessa forma, poupar os pacientes não respondedores de terapias tóxicas e melhorar a sustentabilidade na saúde poupando os custos com drogas não eficientes para um grupo de pacientes.

Rectal cancer is the second most common cancer in large intestine, corresponding to one third of total cases of colorectal cancer (CRC). Patients with rectal cancer in stage II and III are treated with neoadjuvant chemoradiation followed by surgical resection. Analyzes of the resected tumor demonstrated that only 10-45% of the patients achieve pathological complete response (pCR) after neoadjuvant therapy, which is associated with a decrease in local recurrence, improvement of disease free survival and increase in sphincter preservation. Despite the improvement in survival in the last decades, the response to neoadjuvant therapy is still variable and unpredictable, and before the surgery it is not possible to identify and separate clinically the group of patients that will or will not have complete response to neoadjuvant treatment. Moreover, the mechanisms of resistance of rectal tumors to chemoradiation are poorly understood. Thus, the main objective of this work was to identify biomarkers and cellular mechanisms related to the resistance to neoadjuvant therapy in rectal adenocarcinomas and the role of extracellular vesicles (EVs) in this process. The comparative proteomic study between biopsy obtained from tumors pretreatment with residual tumor, post chemoradiation treatment, removed by surgery showed an important alteration in the protein expression profile. Among the proteins with increased expression after neoadjuvant therapy are the DNA repair proteins Ku70 and Ku80, and the protein involved in the intracellular trafficking, Rab5C. It was demonstrated in vitro that Rab5C orchestrates a mechanism of radioresistance in rectal tumors by modulating the EGFR internalization promoted by ionizing radiation (IR). The intracellular EGFR is essential to regulate Ku70 and Ku80 expression and the cell resistance to IR. These data pointed Rab5C and EGFR as potential therapeutic targets to sensitize rectal cancer cells resistant to neoadjuvant treatment. It was also observed that IR promotes epigenetic alterations, predominantly hypomethylation, and between the altered genes are SPG20 and TBC1D16, the first is important to EGFR internalization, while the second regulates Rab5C and modulates EGFR. The protein expression profile was further compared between biopsy pretreatment of patients with and without pCR, and the results showed that these two groups of patients present a different protein expression profile. In patients with pCR the proteins with increased expression are involved in pathways favoring the response to therapy, as glutathione-mediated detoxification and glycogen degradation, while the proteins with increased expression in patients without pCR are involved in tumor energetic metabolism pathways that contribute to tumor resistance to therapy. The observed differences in these groups should be widely explored since they may be predictive markers of response to chemoradiation treatment. The performance of functional studies was possible by generation of a cellular model of rectal tumor resistant to radiotherapy. The analysis of the EVs secreted by these cells showed that IR does not alter the amount and the medium size of secreted EVs, but is able to change their protein content. EVs from irradiated cells presented a different protein profile when compared to EVs from non-irradiated cells, where it was found the increased expression of Ku70, Ku80 and Rab5C, besides the methyltransferases NSUN2 and GLYM in EVs after irradiation. Interestingly, the EVs secreted by irradiated cells are capable of transfering resistance to IR to non-irradiated cells. Moreover, the results showed that the treatment of SW837 cells with EVs from irradiated cells promoted methylation in 98% of the analyzed DNA in comparison with the treatment with EVs from non-irradiated cells. The hypermethylated genes are involved in pathways related to immune system, as antigen presentation, primary immunodeficiency signaling and dendritic cells maturation. Lastly, it was identified that the A33 expression is related to the colorectal tumors differentiation degree, and this protein is present in EVs secreted by rectal adenocarcinoma, indicating that it may be used to isolate EVs specific from colorectal tissues. The data obtained in this work pointed to mechanisms related to resistance to neoadjuvant therapy in rectal adenocarcinoma that together will allow to identify new therapeutic targets with the potential to improve the response to chemoradiation, as well as to identify markers of response to neoadjuvant therapy before the treatment, and, in this way, avoid the non-responder patients to receive toxic therapies and improve health sustainability, sparing cost with non-efficient drugs for a group of patients.

Chemical salivary composition and its relationship with periodontal disease and dental calculus

Aim: To determine the relationship between the chemical composition of saliva, periodontal disease and dental calculus. Methods: An observational analytical cross-sectional study was conducted with patients over 55 years of age. Ethical principles of autonomy and risk protection were applied according to the international standards. Sociodemographic and diagnosis variables (presence of dental calculus and periodontal status) were considered to measure salivary concentrations of glucose (by the glucose oxidase/peroxidase method, amylase (by the colorimetric test), urea (by the amount of indophenol), total protein (by the Bradford method) and albumin (by the nephelometric method). Patients chewed a sterile rubber band and 3 mL of stimulated saliva were collected. The samples were stored at -5 °C, centrifuged at 2,800 rpm for 10 min, and the supernatant was removed and stored at -20 °C. Data were presented as frequencies and proportions for qualitative variables and measures of central tendency and dispersion for quantitative variables. Data were analyzed by either analysis of variance or Kruskal Wallis test . A p value <0.05 was considered statistically significant. Results: Significant relationships were observed between the concentration of salivary urea and periodontal status (p = 0.03) and the presence of dental calculus and urea (p = 0.04) was demonstrated. Conclusions: A relationship between the salivary urea concentration and the presence of periodontal disease and dental calculus is suggested.

A comparison of CA125, HE4, risk ovarian malignancy algorithm (ROMA), and risk malignancy index (RMI) for the classification of ovarian masses

OBJECTIVE: Differentiation between benign and malignant ovarian neoplasms is essential for creating a system for patient referrals. Therefore, the contributions of the tumor markers CA125 and human epididymis protein 4 (HE4) as well as the risk ovarian malignancy algorithm (ROMA) and risk malignancy index (RMI) values were considered individually and in combination to evaluate their utility for establishing this type of patient referral system. METHODS: Patients who had been diagnosed with ovarian masses through imaging analyses (n = 128) were assessed for their expression of the tumor markers CA125 and HE4. The ROMA and RMI values were also determined. The sensitivity and specificity of each parameter were calculated using receiver operating characteristic curves according to the area under the curve (AUC) for each method. RESULTS: The sensitivities associated with the ability of CA125, HE4, ROMA, or RMI to distinguish between malignant versus benign ovarian masses were 70.4%, 79.6%, 74.1%, and 63%, respectively. Among carcinomas, the sensitivities of CA125, HE4, ROMA (pre-and post-menopausal), and RMI were 93.5%, 87.1%, 80%, 95.2%, and 87.1%, respectively. The most accurate numerical values were obtained with RMI, although the four parameters were shown to be statistically equivalent. CONCLUSION: There were no differences in accuracy between CA125, HE4, ROMA, and RMI for differentiating between types of ovarian masses. RMI had the lowest sensitivity but was the most numerically accurate method. HE4 demonstrated the best overall sensitivity for the evaluation of malignant ovarian tumors and the differential diagnosis of endometriosis. All of the parameters demonstrated increased sensitivity when tumors with low malignancy potential were considered low-risk, which may be used as an acceptable assessment method for referring patients to reference centers.

Evolución del estado nutricional en pacientes en hemodiálisis durante 4 años de seguimiento

La malnutrición proteíco calorica asi como la inflamación sistémica y metabólica son trastornos frecuentes entre los pacientes con insuficiencia renal crónica sometidos a tratamiento renal sustitutivo (Hemodiálisis), lo que contribuye a su morbilidad y mortalidad. En este trabajo se ha seguido a 90 pacientes de ambos sexos con insuficiencia renal crónica que fueron tratados con hemodiálisis periódicamente en nuestra unidad durante cuatro años. A todos los pacientes se le realizaron mediciones trimestrales de albúmina plasmática (Alb), colesterol total (CT), proteínas totales (PT) y mensuales de transferrina (Tr), y se les efectuaron mediciones antropométricas de peso, altura e índice de masa corporal calculado mediante la formula peso/talla², y agrupada según la clasificación de la OMS en IMC < 18,50 infrapeso, 18,50 a 24,99 normal, 1,25 a 29,99 sobrepeso y >30 del IMC s/OMS. El objetivo de este trabajo fue evaluar el estado nutricional de estos pacientes mediante la valoración de parámetros bioquímicos y parámetros antropométricos y determinar si estos pacientes sufren alteraciones que sugieran deterioro nutricional directamente relacionado con el tiempo en diálisis. Durante los 4 años todos los pacientes manifestaron un importante descenso de los parámetros bioquímicos, en cambio el IMC no presentó cambios significativos en relación a la desnutrición. Por lo tanto la desnutrición de los pacientes en diálisis es un hecho patente, el IMC no se corresponde con lo parámetros bioquímicos observados, por lo que el deterioro nutricional de estos pacientes se manifiesta principalmente mediante los parámetros bioquímicos estudiados.

Protein-calorie malnutrition as well as systemic inflammation and metabolic disorders are common among patients with chronic renal failure undergoing renal replacement therapy (hemodialysis), which contributes to their morbidity and mortality. This work has followed 90 patients of both sexes with chronic kidney disease who were treated with hemodialysis periodically in our unit for four years. All patients were performed quarterly measurements of plasma albumina (Alb), total cholesterol (TC), total protein (TP) and monthly transferrin (Tr), Anthropometric measurements of height and weight were taken on all patients by using a balance/stadiometer (Perperson 113481); weight was measured in kilograms and height in centimetres. BMI was calculated with this formula: weight/height2 and classified according to the WHO criteria: BMI.

Sistema NADPH oxidasa: nuevos retos y perspectivas / NADPH oxidase system: new challenges and perspectives

El sistema NADPH oxidasa es un complejo multiproteico encargado de producir especies reactivas del oxígeno (ROS, por reactive oxygen species) en diferentes células y tejidos. Es de gran importancia en las células fagocíticas (principalmente neutrófilos y macrófagos) porque participa en la destrucción de microorganismos patógenos, mediante la fagocitosis y la formación de las trampas extracelulares de neutrófilos (NET, por neutrophils extracelular traps), así como en la activación de procesos inflamatorios. Las alteraciones en la producción de ROS por parte de las células fagocíticas a causa de defectos genéticos en los componentes del sistema generan la inmunodeficiencia primaria denominada enfermedad granulomatosa crónica (EGC). Este es un artículo de revisión sobre los componentes del sistema NADPH oxidasa, su distribución celular, mecanismo de activación y acción, así como de las funciones que desempeña en otros tejidos. Además, se revisan los defectos moleculares que llevan a la EGC y el tratamiento de esta, incluyendo la terapia con IFNγ, y finalmente las perspectivas para el estudio del sistema.

The NADPH oxidase system is a multiprotein complex that acts as the main source of reactive oxygen species (ROS) in different cells and tissues. In phagocytic cells (mainly macrophages and neutrophils) it is essential for eliminating pathogenic microorganisms, by phagocytosis and the formation of neutrophil extracellular traps (NETs). It also contributes to inflammatory processes. Genetic defects in the components of the system cause alterations in the production of ROS by phagocytic cells, leading to the primary immunodeficiency known as chronic granulomatous disease (CGD). This is a review article on the components of the NADPH oxidase system, its cellular distribution, activation, mechanisms of action, and roles in other tissues. The different molecular defects that lead to EGC are also reviewed, as well as its treatment, including therapy with IFNγ, and the prospects for the study of the system.

Urinary proteomics evaluation in interstitial cystitis/painful bladder syndrome: a pilot study

PURPOSE: Interstitial cystitis/painful bladder syndrome (IC/PBS) is characterized by chronic pain, pressure and discomfort felt in the pelvis or bladder. An in-depth shotgun proteomics study was carried out to profile the urinary proteome of women with IC/PBS to identify possible specific proteins and networks associated with IC/PBS. MATERIALS AND METHODS: Urine samples from ten female IC/PBS patients and ten female asymptomatic, healthy control subjects were analyzed in quadruplicate by liquid chromatography-tandem mass spectrometry (LC-MS/MS) on a hybrid linear ion trap-orbitrap mass spectrometer. Gas-phase fractionation (GPF) was used to enhance protein identification. Differences in protein quantity were determined by peptide spectral counting. RESULTS: a-1B-glycoprotein (A1BG) and orosomucoid-1 (ORM1) were detected in all IC/PBS patients, and = 60 percent of these patients had elevated expression of these two proteins compared to control subjects. Transthyretin (TTR) and hemopexin (HPX) were detected in all control individuals, but = 60 percent of the IC/PBS patients had decreased expression levels of these two proteins. Enrichment functional analysis showed cell adhesion and response to stimuli were down-regulated whereas response to inflammation, wounding, and tissue degradation were up-regulated in IC/PBS. Activation of neurophysiological processes in synaptic inhibition, and lack of DNA damage repair may also be key components of IC/PBS. CONCLUSION: There are qualitative and quantitative differences between the urinary proteomes of women with and without IC/PBS. We identified a number of proteins as well as pathways/networks that might contribute to the pathology of IC/PBS or result from perturbations induced by this condition.

Influência da função renal no consumo alimentar e no estado nutricional de pacientes com insuficiência renal crônica que iniciam tratamento conservador / Influence of renal function in nutritional status and food consumption of pacients with chronic renal insufficiency treated in a conservative way

Pacientes com insuficiência renal crônica (IRC) apresentam freqüentemente algum grau de desnutrição energético-protéica. Uma das principais causas desta condição é a redução no consumo alimentar. A diminuição do consumo de energia e proteínas se deve principalmente à anorexia, freqüentemente observada nesses pacientes. Como a anorexia é um sintoma subjetivo, uma maneira de avaliar sua presença é por meio do consumo alimentar em relação ao nível de função renal tornando assim possível determinar a influência de uremia sobre a anorexia. Assim, o objetivo deste estudo foi o de analisar o consumo alimentar espontâneo e o estado nutricional de pacientes com (IRC) no início do tratamento conservador em relação ao nível de função renal. Para tal, foi analisado um banco de dados que compreendeu um período de 10 anos e foram estudados 487 pacientes com IRC na fase de tratamento conservador, quando de sua primeira avaliação nutricional, e continha informações da primeira consulta sobre ingestão alimentar e parâmetros antropométricos. Os seguintes parâmetros foram analisados: consumo de energia e proteína obtido a partir do registro alimentar de 3 dias, parâmetros antropométricos e albumina sérica. A ingestão de proteína também foi estimada por meio do PNA, que é o aparecimento de nitrogênio uréico equivalente à proteína. Na análise dos dados demográficos dessa população, observamos predominância do gênero masculino (61,6%), e os pacientes se encontravam em média na quinta década de vida (53,4±15,0 anos). Observamos como principal causa da IRC o diabetes mellitus (27,7%), seguido da nefropatia hipertensiva (25,2%). O clearance de creatinina médio foi de 34,1±18,8 mLmin/1,73 m². O consumo médio de energia foi de 24,9±8,2 Kcal/Kg/dia e o de proteínas foi de 0,95±0,36 g/Kg/dia. O valor médio do PNA foi de 0,98±0,32 g/Kg/dia e a albumina média foi de 3,8±0,7 g/dL. A adequação de peso e o índice de massa corporal...

Efeitos renais crônicas em trabalhadores expostos ao chumbo e suas relaçöes com a pressäo arterial / The chronic renal effects in workers occupationally exposed to lead and its correlation with arterial pressure

Determinaram-se as atividades de duas enzimas de membrana: alanina-aminopeptidase (AAP), -glutamil-transpeptidase e da enzima lisossomal N-acetil-B-D-glucosaminidase, os níveis de proteína total, albumina e ácido delta-aminolevulínico em urinas de indivíduos expostos ocupacionalmente ao chumbo e de indivíduos näo expostos. Todos os indivíduos apresentavam creatinina sérica inferior a 1,5 mg/dl. Foram determinados os níveis sangüíneos de chumbo e aferidas pressäo arterial diastólica e pressäo arterial sistólica. Objetivou-se investigar a toxicidade renal crônica do chumbo e sua possível correlaçäo com a pressäo arterial em indivíduos com funçäo renal normal. A pressäo arterial diastóeca foi maior no grupo exposto. Observaram-se correlaçöes das pressöes com tempo de exposiçäo e com chumbo no sangue

Estudo das gorduras totais, valor calórico total e proteinas totais no colostro de puérperas, segundo o nível sócio-econômico, o estado nutricional e a paridade / Study of the total fats, total value caloric and total proteins in colostrum of puerperas, according to social-economic level, the nutritional state and the parity

As concentraçöes de Gorduras Totais, Valor Calórico Total e Proteínas Totais do Colostro, obtido de puérperas primípara e multíparas de baixo e alto nível sócio-econômico eutróficas e desnutridas, foram determinadas com o propósito de se conhecer a possível influência das condiçöes maternas (condiçäo nutricional, nível sócio-econômico e paridade) sobre a qualidade do colostro. Näo foram encontradas diferenças significantes do conteúdo desses elementos entre os grupos de puérperas estudadas. Com base no presente estudo, conclui-se que as puérperas podem manter um aleitamento natural independentemente do seu nível sócio-econômico, do seu estado nutricional e da sua paridade

Proteinas séricas en niños de diferentes medios socio-económicos / Blood protein in child from different socioeconomic rules

En el estudio realizado sobre proteinas totales, albúminas, globulinas, relación alb./glab. y nitrógeno de los alfa aminoácidos en niños pertenecientes a distintos estractos sociales de la ciudad de Chosica y del Departamento de Lambayeque, encontrándose que los valores medios hallados está dentro de los valores normales dados por el método utilizado. Al efectuar la comparación estadística de los grupos estudiados, se obtuvo valores mayores en los niños de los pueblos jóvenes que en las ciudades de Chosica y Lambayeque, encontrándose una diferencia significativa en el índice de consumo de proteinas.