In vitro release mechanism and cytotoxic behavior of curcumin loaded casein nanoparticles

Abstract In the recent past, drug delivery through nanoparticles is considered an effective tool to treat various diseases. Biopolymeric nanoparticles such as protein based nanoparticles have vital role as drug carrier as it is non-antigenic, and easily biodegradable. Curcumin, plant polyphenolic anticancerous compound was loaded into the casein nanoparticles by coacervation method. Particle size and surface charge of spherical casein nanoparticles as observed to be 201.4 nm and -86.9 mV. The loading efficiency of curcumin loaded casein nanoparticles was found to 85.05 %. In vitro drug release was performed at different pH (7.4 and 3.0), and the cumulative release was observed to be 24.8 and 20.13% and at different temperatures (25°C and 37°C), the cumulative release was observed to be 24.8 and 28.60 % respectively in 48 h. Curcumin release from casein nanoparticles was shown to be in a steady, and prolonged rate. The nanoparticles were observed to have an effective antimocrobial activity than curcumin in free form. The drug loaded casein nanoparticles were found to be potent particles to protect cells from hydrogen peroxide and UV light damage. The cytotoxic activity of nanoparticles on MCF7 and A549 cells were assayed and was observed to have an IC50 value of 609 and 825.2µg/ml. Cell death was observed to be through apoptosis, accompanied by DNA fragmentation.

Análisis genético de cinco polimorfismos de nucleótido simple de caseínas lácteas obtenidos con chips genómicos en ganado Holstein de Antioquia, Colombia / Genetic analysis of five single nucleotide polymorphisms of dairy caseins obtained with genomic chips in Holstein cattle from Antioquia, Colombia

RESUMEN Los polimorfismos genéticos asociados con las caseínas de la leche son de gran importancia, ya que pueden ser usados como marcadores genéticos para mejorar el rendimiento productivo en los hatos lecheros. El objetivo de este estudio fue evaluar la diversidad y estructura genética de 5 SNP de caseínas de la leche, obtenidos con chips genómicos en vacas y toros de raza Holstein en Antioquia (Colombia). Fueron muestreados 113 animales de raza Holstein en 3 regiones del departamento de Antioquia (norte, centro y oriente) y un cuarto grupo de sementales comerciales. Los animales fueron genotipificados con chips genómicos de alta densidad (Illumina BovineHD e Illumina SNP50 v2), a partir de los cuales se identificaron 5 SNP (ARS-BFGL-NGS-8140, BTA-77380-no-rs, BTA-32346-no-rs, BTB-00821654 y ARS-BFGL-NGS-15809). Para cada SNP se realizó un análisis genético mediante un análisis de varianza molecular (amova) usando el software GenAIEx 6.501. Los SNP con mayor heterocigosidad total (HT) fueron ARS-BFGL-NGS-8140 y BTA-32346-no-rs, con resultados cercanos al 45%; sin embargo, la Ht para ARS-BFGL-NGS-15809, BTA-77380-no-rs y BTB-00821654 estuvo por debajo del 15%. El SNP con mayor diversidad genética fue BTA-32346-no-rs (Ho-He = 0,06; p < 0,05). En esta investigación se evaluó una subpoblación de toros comerciales extranjeros, en la cual se obtuvieron frecuencias alélicas y genotípicas similares a las obtenidas para las subpoblaciones locales, sugiriendo que los alelos de los toros muy posiblemente están fijados en dichas subpoblaciones, por lo que la estructura y diversidad genética tienden a ser bajas en la muestra de estudio.

ABSTRACT Genetic polymorphisms associated with milk caseins have a great importance since they can be used as genetic markers to improve productive performance in dairy herds. The main goal of the present study was to evaluate the diversity and genetic structure of 5 SNPs of milk caseins, obtained with genomic chip in Holstein cows and bulls from Antioquia (Colombia). 113 Holstein animals were sampled in 3 regions of Antioquia (north, center, and east), and a fourth group of commercial sires. Animals were geno-typed with high-density SNP chips (Illumina BovineHD and Illumina SNP50 v2), from which 5 SNPs were identified (ARS-BFGL-NGS-8140, BTA-77380-no-rs, BTA-32346-no-rs, BTB-00821654 and ARS-BFGL-NGS-15809). For each SNP, a genetic analysis was performed by means of an analysis of molecular variance (AMOVA) using the GenAIEx 6.501 software. The SNPs with the highest total heterozygosity (Ht) were ARS-BFGL-NGS-8140 and BTA-32346-no-rs, with results close to 45%; however, the HT for ARS-BFGL-NGS-15809, BTA-77380-no-rs, and BTB-00821654 were below 15%. The SNP with the highest genetic diversity was BTA-32346-no-rs (Ho-He = 0,06; p < 0,05). In this research a subpopulation of foreign commercial bulls was evaluated, in which similar allelic and genotypic frequencies to those for local subpopulations were obtained, suggesting that the alleles of the bulls are very possibly fixed in these subpopulations, so that the structure and genetic diversity tend to be low in the study sample.

Validation of a method employing liquid chromatography to identify and quantify glycomacropeptide of casein in raw milk / Validación de un método mediante cromatografía líquida para identificar y cuantificar glicomacropéptido de caseína en la leche cruda

Background: The information on official testing methods, or regulatory methods in Colombia to test whey in milk is limited; this restriction of information goes against the possibility of mitigating the risk of food fraud. Objectives: The validation of an HPLC method to determine casein glycomacropeptide (c-GMP), a protein that countries such as Brazil, Spain, and Ecuador have used as an indicator of raw milk adulteration with whey, was carried out. Methods: A 10mL sample of raw milk is precipitated with 24% TCA using ultrasound, a process followed by filtration. The collected fraction ensured the separation of c-GMP and then injected into the liquid chromatography. Results: A 30 minutes analysis allowed the determination of c-GMP with a retention time of 12.9 ± 0.5 minutes. The performance characteristics method in the validation exercise were: recovery percentage 99.97%, linearity R2> 0.95; % RSD accuracy <5.3%. Conclusion, the method exhibits desirable attributes for the intended purpose

Antecedentes: En Colombia la información de dominio público en metodologías de análisis de lactosuero en leche es limitada, restringiendo la posibilidad de acceder a ellas para mitigar el riesgo de fraude alimentario. Objetivos: Se realizó validación de un método por HPLC para determinar en leche cruda c-GMP, proteína usada como indicador de adulteración en países como Brasil y Ecuador. Metodos: Una muestra de 10mL de leche cruda es precipitada con TCA al 24% empleando ultrasonido, proceso seguido por filtración. La fracción recolectada aseguró la separación del c-GMP para luego inyectar al cromatógrafo líquido. Resultados: La determinación de c-GMP permitió el análisis en 30 minutos con tiempo de retención de 12,9 ± 0,5 minutos. Las características de desempeño del método en el ejercicio de validación fueron: porcentaje de recuperación 99,97%, linealidad R2>0,95; precisión %RSD< 5,3%. Conclusión: el método al final del ejercicio exhibe atributos para el fin previsto

Biomateriales que inducen la remineralización del esmalte dental y dentina / Biomaterials that induce remineralization of the dentin and enamel surface

Desde los sesentas, con la invención del vidrio bioactivo, los tratamientos de remineralización se han popularizado entre los cirujanos dentistas y su utilización es cada vez mayor; la remineralización, en conjunto con las adecuadas medidas de higiene preventiva, representa uno de los mejores abordajes mínimamente invasivos y a un costo comparativamente bajo. Este estudio documental tiene por objetivo establecer una mejor comprensión del uso clínico de los biomateriales que inducen la remineralización de la superficie del esmalte dental y dentina. Se realizó una exploración utilizando motores de búsqueda (bases de datos en PubMed, Medigraphic, y Science Direct). El proceso de localización de los estudios relevantes se efectuó introduciendo palabras clave como: silicatos de calcio, fosfopéptidos de caseína-fosfato de calcio amorfo, remineralización, esmalte y dentina, incluyéndose en el procedimiento artículos de antigüedad no superior a siete años, en español e inglés, publicados en revistas científicas aprobadas por pares.Actualmente, no es posible remineralizar del todo la estructura dentaria, por lo cual, en un futuro cercano, los esfuerzos de la odontología de remineralización deben apuntar al desarrollo de agentes biomiméticos inteligentes que restauren al cien por ciento la estructura dental perdida (AU)

Since the sixties, with the invention of bioactive glass, remineralization treatments have become popular among dental surgeons. Their usage is increasing; remineralization, in conjunction with appropriate preventive hygiene measures, represents one of the best minimally invasive treatments at a relatively low cost. This documentary study aims to establish a better understanding of the clinical use of biomaterials that induce remineralization of the surface of teeth enamel and dentin. A search was conducted using search engines (PubMed and Medigraphic databases, and Science Direct). The search process for the relevant studies was carried out by introducing keywords such as calcium silicates, phosphopeptides of amorphous calcium casein-phosphate, remineralization, enamel and dentin, including in the search articles no older than seven years in Spanish and English published in scientific reviewed journals. Currently, it is not possible to completely remineralize the dentary structure so, in the near future, remineralization dentistry efforts should aim to develop (AU)

Allele and genotype frequency for milk beta-casein in dairy cattle in the northern region of Tocantins State, Brazil / Frequências alélicas e genotípicas para beta-caseína do leite em bovinos leiteiros da microrregião de Araguaína, Tocantins

At present, there is a concern about the quality of milk and diseases related to its consumption, as it can generate discomfort and allergic reactions in some individuals due to its protein components. Thus, the present study was developed to identify the allele and genotype frequencies of genes for ß-casein, A1 and A2, in dairy herds in the region of Araguaína-TO, Brazil. Genetic material from 421 animals (crossbred dairy cattle in lactation) was used. All animals were numbered for identification, and DNA samples were extracted from hair bulbs. Samples for two markers from the polymorphic regions were characterized and confirmed by real-time PCR using the ABI Prism® 7500 Sequence Detection System (Applied Biosystems). Allele and genotype frequencies were determined using the TaqMan™ detection system, where the primer and probe release different fluorescence signals for each allele of the polymorphism. The sampled herd showed frequencies of 28.27% for the A1 allele and 71.73% for the A2 allele. Genotype frequencies were 52.96% (223/421) for A2A2; 37.53% (158/421) for the A1A2 genotype; and 9.50% (40/421) for the A1A1 genotype. The frequency of the A1 allele for ß-casein in dairy herds from the northern region of Tocantins was low and is per the results of previous studies. Although the A2A2 genotype of ß-casein had a high relative frequency, the A1A2 genotype is still rather frequent, warranting greater selection pressure.(AU)

Atualmente existe uma preocupação em relação à qualidade e doenças que estão relacionadas ao consumo de leite, pois o mesmo pode gerar desconfortos e reações alergicas em alguns indivíduos devido aos seus constituintes protéicos. Assim, o presente estudo teve como objetivo identificar a frequência alélica e genotípica de genes para beta caseína, A1 e A2, em rebanhos leiteiros da região de Araguaína-TO. Foram utilizados material genético de 421 animais (bovinos leiteiros mestiços em lactação), e todos os animais foram numerados para identificação e amostras de DNA foram extraídas de bulbo de folículos pilosos. As amostras para dois marcadores das regiões polimórficas foram caracterizadas e confirmadas por PCR em tempo real, usando um sistema de detecção de sequências ABI Prism® 7500 (Applied Biosystems). As frequências alélicas e genotípicas foram determinadas utilizando o sistema de detecção TaqMan ™, no qual o primer e a sonda emitem diferentes sinais de fluorescência para cada alelo do polimorfismo. Observou-se frequência do alelo A1 de 28,27%, e do alelo A2 de 71,73% no rebanho amostral. A frequência genotípica de A2A2 foi de 52,96% (223/421), com genótipo A1A2 de 37,53% (158/421), e de 9,50% (40/421) animais com genótipo A1A1. A frequência do alelo A1 para beta-caseína em rebanhos leiteiros da região norte do Tocantins foi baixa e seguiu a mesma tendência já observada em estudos anteriores. Os genótipos A2A2 da beta-caseína apresentaram frequência relativa alta, entretanto o genótipo A1A2 ainda é bastante frequente, necessitando de maior pressão de seleção.(AU)

Influencia de la suplementación con vitamina D, omega 3 o el uso de una dieta libre de gluten y caseína en el comportamiento de niños y niñas con trastorno del espectro autista / Influence of the vitamin D, omega 3 supplementation or the use of a gluten and casein free diet in the behavior of boys and girls with autism spectrum disorder

La prevalencia de trastorno del espectro autista ha ido en aumento, sin embargo, en Chile no existen lineamientos nutricionales acerca del posible tratamiento de la condición. Es por ello que el objetivo de esta revisión fue analizar la evidencia actual en relación al uso de una dieta libre de gluten y caseína, suplementación de vitamina D y omega 3 y su impacto en el comportamiento de niños/as con trastorno del espectro autista. Hay evidencia con resultados en torno a los beneficios de la suplementación con vitamina D debido a su carácter neuroprotector y su función neuromuscular. A su vez, la evidencia con omega 3 (DHA) es estadísticamente significativa para irritabilidad, hiperactividad, letargo, comportamiento estereotipado, conciencia social, comunicación y disminución de la severidad del autismo. Respecto a la dieta libre de gluten y caseína lo observado es que no existe evidencia que respalde los beneficios que esta exclusión entregaría. Aún falta evidencia para declarar un manejo nutricional específico para el tratamiento de los síntomas gastrointestinales y de comportamiento, más allá de la suplementación con aquellos micronutrientes en déficit.

The prevalence rate of autism spectrum disorder has been increasing, however, in Chile there are no nutritional guidelines about the possible treatment of the condition. That is the reason why the aim of this review is to analyze the current evidence regarding the use of a gluten and casein free diet, vitamin D and omega 3 supplementation and its impact in the behavior of children's with spectrum disorder autistic. There is evidence based on significant results regarding the benefits of vitamin D supplementation due to its neuroprotective character and neuromuscular function. At the same time the omega 3 evidence is statistically significant in the diminution of irritability hyperactivity, lethargy, stereotypical behavior, severity of autism and increase of social consciousness and communication. Regarding the gluten-free and casein-free diet, what was observed is that there is no evidence to support the benefits that this exclusion would provide. The evidence has not been conclusive to declare a specific nutritional management for the treatment of gastrointestinal and behavioral symptoms, beyond supplementation with those micronutrients in deficit.

Comparative effects of cpp-acp and xylitol f-varnishes on the reduction of tooth erosion and its progression

Abstract Evaluated the effect of CPP-ACP/NaF and xylitol/NaF varnishes in reduce erosion and progression of erosion. Forty enamel blocks were divided into four groups (n=10): G1=CPP-ACP/NaF varnish (MI varnishTM); G2=xylitol/NaF varnish (Profluorid®); G3=NaF varnish (Duraphat®, positive control) and G4=deionized water (MilliQ®, negative control). Samples were immersed in Sprite ZeroTM (pH 2.58, 4x/day, 3 days), in between immersions, the specimens stayed in artificial saliva. After 3 days of erosion, the eroded area was divided in two (half of one received an additional varnish layer while the other half repeated the same 3-day erosion cycle). The 3D, non-contact profilometry technique was used to determinate tooth structure loss (TSL) and surface roughness (SR). Scanning electron microscopy (SEM) and 3D images were utilized to evaluate the topography of the samples. Mann-Whitney, one-way ANOVA and Tukey tests were used (significance level of 0.05%). SEM and 3D images were descriptively evaluated. After 3 or 6 days of erosion, all tested varnishes were better than G4 (p<0.05) for TSL and SR. In addition, G1 had lower values for TSL than G3 (p<0.05) after 3 days of erosion. Under SEM and 3D images observation, all groups presented porosity, irregularities and depressions on the surface enamel after 3 and 6 days of erosion, more pronounced in G4. An application of topical NaF varnishes was effective in reducing TSL and enamel roughness after erosion challenges, being the CCP-ACP/NaF varnish more effective than NaF varnish and water after 3 days of erosion.

Resumo Avaliou-se o efeito dos vernizes CPP-ACP/NaF e xilitol/NaF na redução da erosão e progressão da erosão. Quarenta blocos de esmalte foram divididos em quatro grupos (n=10): G1=verniz CPP-ACP/NaF (verniz MITM); G2=verniz xilitol/NaF (Profluorid®); G3=verniz NaF (Duraphat®, controle positivo) e G4=água desionizada (MilliQ®, controle negativo). As amostras foram imersas em refrigerante Sprite ZeroTM (pH 2.58, 4x/dia, 3 dias), entre imersões, os espécimes ficaram em saliva artificial. Após 3 dias de erosão, a área erodida foi dividida em duas (metade recebeu uma camada adicional de verniz, enquanto a outra metade repetiu o mesmo ciclo de erosão de 3 dias). A técnica de perfilometria 3D de não contato foi utilizada para determinar a perda de estrutura dentária (PED) e rugosidade superficial (RS). Microscopia eletrônica de varredura (MEV) e imagens em 3D foram utilizadas para avaliar a topografia das amostras. Testes de Mann-Whitney, One-way ANOVA e Tukey foram utilizados (nível de significância de 0,05%). Imagens do MEV e 3D foram avaliadas descritivamente. Após 3 ou 6 dias de erosão, todos os vernizes testados foram melhores que G4 (p<0,05) para PED e RS. Além disso, o G1 apresentou menores valores de PED do que o G3 (p<0,05) após 3 dias de erosão. Observando as imagens em MEV e 3D, todos os grupos apresentaram porosidade, irregularidades e depressões no esmalte superficial após 3 e 6 dias de erosão, sendo mais pronunciados no G4. Uma aplicação tópica de vernizes fluoretados foi eficaz na redução da rugosidade e PED do esmalte após desafios de erosão. Além disso, o grupo CPP-ACP/NaF teve melhor desempenho na redução da PED quando comparado ao verniz de NaF e a água, após 3 dias de erosão.

Comparative effect of calcium mesoporous silica versus calcium and/or fluoride products against dental erosion

Abstract The aim of this study was to compare the effects of a single application of a new calcium mesoporous silica nanoparticle (Ca2+-MSN) versus other calcium and/or fluoride products against dental erosion. Enamel blocks were half-covered and assigned to six groups (n = 10): Ca2+-MSNs; casein phosphopeptide-amorphous calcium phosphate mousse (CPP-ACP); CPP-ACP/F− (900 ppm F−); titanium tetrafluoride (TiF4 1%); sodium fluoride (NaF 1.36%); and Milli-Q® water (negative control). A single application for each product was completed on the exposed areas of the blocks and were submitted to an erosive challenge. Differences in volumetric roughness (Sa), and tooth structure loss (TSL) by use of three-dimensional noncontact optical profilometry were evaluate. Scanning electron microscopy (SEM) was performed. Analysis of variance and Tukey's test for Sa and the Kruskal-Wallis and Mann-Whitney U tests (p< 0.05) for TSL, respectively. Results: When evaluating Sa, all products presented differences in roughness when compared with the control group (p< 0.05) but not with each other (p > 0.05). However, when analyzing the TSL, it was observed that Ca2+-MSNs, TiF4, and NaF were more effective in preventing dental erosion versus CPP-ACP, CPP-ACP/F−, and Milli-Q® water (p< 0.05). In the SEM images, the negative control presented the worst loss of dental structure, with more porous enamel. Ca2+-MSNs were as effective as TiF4 and NaF to reduce the tooth structure loss.

Resumo O objetivo deste estudo foi comparar os efeitos de uma única aplicação de uma nova nanopartícula de sílica mesoporosa de cálcio (Ca2+ -MSN) versus outros produtos à base de cálcio e / ou fluoreto contra a erosão dentária. Blocos de esmalte foram parcialmente cobertos e distribuídos em seis grupos (n = 10): Ca2+ -MSNs; fosfopeptídeos de caseína/fosfato de cálcio amorfo (CPP-ACP); CPP-ACP / F- (900 ppm F-); tetrafluoreto de titânio (TiF4 1%); fluoreto de sódio (NaF 1,36%); e água Milli-Q® (controle negativo). Uma única aplicação para cada produto foi realizada nas áreas expostas dos blocos e submetida a desafio erosivo. Diferenças na rugosidade volumétrica (Sa) e na perda de estrutura dentária (TSL) por meio de perfilometria tridimensional de não contato foram avaliadas. Microscopia eletrônica de varredura (MEV) foi realizada. Foram realizadas análise de variância e teste de Tukey para os testes Sa e Kruskal-Wallis e Mann-Whitney (p<0,05) para TSL, respectivamente. Na avaliação de Sa, todos os produtos apresentaram diferenças de rugosidade quando comparados ao grupo controle (p<0,05), mas não entre si (p> 0,05). No entanto, ao analisar o TSL, observou-se que Ca2+ -MSNs, TiF4 e NaF foram mais eficazes na prevenção da erosão dental versus CPP-ACP, CPP-ACP / F- e Milli-Q® (p<0,05). Nas imagens de MEV, o controle negativo apresentou a pior perda de estrutura dentária, com o esmalte mais poroso. A Ca2+ -MSNs foi tão eficaz quanto o TiF4 e o NaF para reduzir a perda da estrutura dentária.

Índice de CMP em leite pasteurizado comercializado em Minas Gerais, Brasil, durante os anos de 2011 a 2017 / Caseinomacropeptide index in pasteurized milk retailed in Minas Gerais, Brazil from 2011 to 2017

Cheese whey is a nutritious byproduct in the dairy industry, however, due to low commercial value, its use as a milk adulterant is a common practice not easily detected by routine analysis. In Brazil, quantification of caseinomacropeptide (CMP) index, using High Performance Liquid Chromatography (HPLC), is officially used to investigate illegal cheese whey addition to milk. Milk with CMP index above 30mg/L is considered not suitable for human consumption. The objective of this research was to report the CMP index in 185 samples of pasteurized milk, representing 73 commercial brands produced in 51 counties and ten mesoregions of the state of Minas Gerais, from 2011 to 2013 (58 samples) and 2015 to 2017 (127 samples). CMP index was considered normal (up to 30mg/L) in 75.1% of the samples. However, 21.1% presented CMP index above 75mg/L and 3.8% from 31 to 75mg/L. CMP index above 75mg/L was found in 17.4% of the samples produced during the dry season (April to September) and in 24.7% during the rainy season (October to March). These data point to the need of more efficient monitoring and inspection processes to hinder adulteration with cheese whey addition to milk.(AU)

Effect of CPP-ACP on remineralization of artificial caries-like lesion: an in situ study

Abstract The purpose of this double-blind, randomized, crossover in situ study is to compare remineralization of preformed enamel lesions by casein phosphopeptide-stabilized amorphous calcium phosphate (CPP-ACP) and fluoride dentifrice products. During each of four 10-day experimental legs, 10 participants wore intraoral removable palatal acrylic appliances with four human enamel slabs with preformed lesions. A 0.03-mL treatment paste was dripped extraorally onto the enamel blocks once a day for 3 min. The four randomly allocated treatments were as follows: CO- Control: silica dentifrice without fluoride; MP: MI Paste; MPP: MI Paste Plus and FD: Fluoride dentifrice - 1100 ppm F as NaF). Knoop surface hardness (SH) test was performed in three stages (T0 - sound enamel, T1 - after preformed lesion, and T2 - after treatment) and the cross-sectional hardness (CSH) test was performed after treatment using a 50-gram Knoop load for 15 s. Knoop hardness number (KHN) was similar between treatments. %SHr was significantly higher in the MP, FD, and MPP when compared to CO group (Kruskal-Wallis and Mann-Whitney tests, p < 0.05). Harder enamel was found in MP (75 μm) and FD groups at 75 to 175 μm. Treatment with DF, MP, and MPP promoted an increase of 20.27%, 19.24%, and 14.71%, respectively, in Integral Hardness Change (ΔIHC) when compared to CO (p<0.05). Remineralizing agents (MP, MPP, and DF) were able to inhibit demineralization of human enamel subjected to high cariogenic challenge in situ. DF had the greatest preventive potential against the progression of carious lesions.

The use of a new calcium mesoporous silica nanoparticle versus calcium and/or fluoride products in reducing the progression of dental erosion

Abstract Objective There is increasingly common the consumption more times a day of foods and acidic drinks in the diet of the population. The present study aimed to evaluate and compare the effects of a calcium mesoporous silica nanoparticle single application of other calcium and/or fluoride products in reducing the progression of dental erosion. Methodology Half of the eroded area was covered of 60 blocks of enamel, after which the block was submitted to the following treatments: (Ca2+-MSN), casein phosphopeptide-amorphous calcium phosphate (CPP-ACP); CPP-ACP/F-(900 ppm F−); titanium tetrafluoride (TiF4 1%) (positive control); sodium fluoride (NaF 1.36%) (positive control); and Milli-Q® water (negative control) before being submitted to a second erosive challenge. A surface analysis was performed via a three-dimensional (3D) noncontact optical profilometry to assess the volumetric roughness (Sa) and tooth structure loss (TSL) and and through scanning electron microscopy (MEV). An analysis of variance (ANOVA) and Tukey's test were performed. Results Regarding Sa, all experimental groups exhibited less roughness than the control (p<0.05). The TSL analysis revealed that the Ca2+-MSN and NaF groups were similar (p>0.05) and more effective in minimizing tooth loss compared with the other groups (p<0.05). Conclusions The Ca2+-MSN and NaF treatments were superior compared with the others and the negative control.

Autism spectrum disorder: a systematic review about nutritional interventions / Transtorno do espectro autista: uma revisão sistemática sobre intervenções nutricionais

ABSTRACT Objective: To identify and analyze the scientific evidence of nutritional interventions performed in children and adolescents with Autism Spectrum Disorder. Data sources: A systematic review was conducted in the MEDLINE, Cochrane Library, Embase, LILACS, Google Scholar, PubMed, PsycINFO and Periódicos CAPES databases, using a search strategy to identify studies published between January 2003 and March 2018, in Portuguese, English and Spanish. Were included studies that described nutritional interventions in children and adolescents with autism spectrum disorders and assessed autistic behavior and/or gastrointestinal symptoms. We excluded other review articles and studies that did not include a control group in the research design. The studies were reviewed for descriptive information, and the quality of evidence was assessed through the GRADE system. Data synthesis: 18 studies were included in the review, being 16 randomized clinical trials, 1 case-control study and 1 open-label trial. As a result, the implementation of a gluten-free and casein-free diet was the most used intervention among the studies. Of the total, 10 studies showed a positive association of intervention with the evaluated results, while 8 did not find of a significant association. Conclusions: Although some authors report progress in the symptoms associated with autism in individuals with Autistic Spectrum Disorder undergoing nutritional interventions, there is little scientific evidence to support the use of nutritional supplements or dietary therapies in children and adolescents with autism.

RESUMO Objetivo: Identificar e analisar as evidências científicas de intervenções nutricionais realizadas em crianças e adolescentes com Transtorno do Espectro Autista. Fontes de dados: Realizou-se uma revisão sistemática nas bases de dados MEDLINE, Cochrane Library, Embase, LILACS, Google Acadêmico, PubMed, PsycINFO e Periódicos CAPES utilizando estratégia de busca abrangente para identificar estudos publicados entre janeiro de 2003 e março de 2018, em língua portuguesa, inglesa e espanhola. Foram incluídos estudos que descreveram intervenções nutricionais em crianças e adolescentes com Transtorno do Espectro Autista e avaliaram sintomas comportamentais e/ou sintomas gastrintestinais, sendo excluídos artigos de revisão e estudos que não incluíram um grupo controle em seu delineamento. Os estudos foram examinados para obter informações descritivas, e a qualidade de evidência foi avaliada por meio do Sistema GRADE (Grading of Recommendations Assessment, Development and Evaluation). Síntese dos dados: Dezoito estudos foram incluídos na revisão (16 ensaios clínicos randomizados, um estudo de caso-controle e um ensaio clínico aberto). As intervenções e os resultados variaram, entretanto a implementação de uma dieta livre de glúten e caseína foi a intervenção mais utilizada entre os estudos. Do total, dez estudos encontraram associação positiva entre intervenção e resultados avaliados, enquanto oito não encontraram associação significativa. Conclusões: Embora alguns autores exponham progressos nos sintomas associados ao autismo em indivíduos com esse transtorno submetidos a intervenções nutricionais, há poucas evidências científicas para apoiar o uso destas em crianças e adolescentes com autismo.

Effect of fermented milk with kefir grains on the in vitro demineralization of bovine tooth enamel / Efecto de la leche fermentada con granos de kéfir sobre la desmineralización in vitro del esmalte dental bovino

The dental caries is a progressive destruction of the teeth tissue due to the disbalance in the normal molecule interactions between the enamel and the bio!lm, which alters the demineralization-remineralization process. Milk fermentation produces caseinphosphopeptides with proved remineralizing capacity of the enamel. The presence of these peptides in fermented milk with ke!r grains has been described. The purpose of this work was to evaluate in vitro the capacity of milk ke!r to prevent the demineralization of dental enamel. Bovine incisors (n=68, 17 per group) were treated for 72 h with different solutions: I: artificial saliva at pH 7.2 , II: demineralizing solution at pH 4.5, III: supernatant of kefir fermented milk at pH 4.5, IV: milk supernatant at pH 4.5. The effects of treatments were evaluated by the change in the weight of the specimens, calcium concentration in the solution and by scanning electron microscopy (SEM) of the enamel. Kefir milk supernatant prevented the demineralization process, that was evidenced by a change in weight and calcium concentration that were not different from group I, although the pH was 4.5. In contrast, group IV showed a decrease in weight and an increase in calcium concentration, compared with group I (one way ANOVA, p<0.05). Images of SEM agree with the values of weight and calcium concentration. These results indicate that kefir milk supernatant has a protective effect on enamel demineralization in vitro. (AU)

La caries dental es una patología debido a un desequilibrio en las interacciones moleculares normales entre el esmalte y la biopelícula, que altera el proceso de desmineralización remineralización. La fermentación de la leche produce fosfopéptidos de caseína con probada capacidad remineralizante del esmalte, y se ha descripto la presencia de estos péptidos en la leche fermentada con granos de kéfir. El propósito de este trabajo fue evaluar in vitro la capacidad del kéfir de leche para prevenir la desmineralización del esmalte dental. Sesenta y ocho incisivos bovinos (17 por grupo) fueron tratados durante 72 h con diferentes soluciones: I: saliva artificial, pH 7.2, II: solución desmineralizante, pH 4.5, III: sobrenadante de leche fermentada con kefir, pH 4.5, IV: sobrenadante de leche, pH 4.5. El proceso de desmineralización se evaluó mediante el cambio en el peso de las muestras, la concentración de calcio en la solución y microscopía electrónica de barrido (SEM) del esmalte. El sobrenadante de leche fermentada con kéfir impidió el proceso de desmineralización, que se evidenció por un cambio en el peso y la concentración de calcio que no discreparon del grupo I, a pesar de haber tenido un pH de 4.5. En contraste, el grupo IV mostró una disminución en el peso y un aumento en la concentración de calcio, en comparación con el grupo I (ANOVA a un criterio, p<0.05). Las imágenes SEM concuerdan con los cambios en el peso y la concentración de calcio en los grupos estudiados. Los datos obtenidos demuestran que el sobrenadante de la leche tratada con kéfir tiene un efecto protector sobre la desmineralización del esmalte in vitro, inducida por el pH ácido. (AU)

El caseinato de sodio y la caseína α inhiben la proliferación de la línea celular mieloide de ratón 32D clone 3 (32Dcl3) mediante el TNF-α / Sodium caseinate and alfa-casein inhibit proliferation of the mouse myeloid cell line 32D clone 3 (32Dcl3) via TNF-α

Resumen Introducción. Se ha demostrado que el caseinato de sodio y sus componentes (caseínas α, β y κ) inhiben la proliferación de la línea celular hematopoyética de ratón 32D clone 3 (32Dcl3) e inducen su diferenciación hacia macrófagos. Se sabe que la caseína α induce la producción de IL-1β y que esta última citocina inhibe la proliferación celular mediante la producción del factor de necrosis tumoral alfa (TNF-α), pero se desconoce si el caseinato de sodio y las caseínas inducen la producción de TNF y si este es el responsable de la inhibición de la proliferación. Objetivo. Evaluar si el caseinato de sodio y las caseínas α, β y κ inhiben la proliferación de la línea celular 32Dcl3 mediante la producción de TNF-α. Materiales y métodos. Se usaron diferentes concentraciones de caseinato de sodio y de las caseínas α, β y κ en las células 32Dcl3. Posteriormente, se evaluaron la viabilidad celular mediante una prueba con el MTT [3-(4,5-dimetiltiazol-2-ilo)-2,5-difeniltetrazol], la inducción de apoptosis con la citometría de flujo y la síntesis del TNF-α con el ELISA. Además, se hicieron pruebas de neutralización con anti-TNF-α en células 32Dcl3 tratadas con caseinato de sodio y caseína α, y se evaluó la proliferación celular. Resultados. Se encontró que el caseinato de sodio y las caseínas α, β y κ reducían la proliferación de la línea celular 32Dcl3 sin afectar la viabilidad, y que solo el caseinato y la caseína α inducían la apoptosis y la liberación al medio de TNF-α. La proliferación de células 32Dcl3 tratadas con caseinato y caseína α se restableció al usar anticuerpos anti-TNF-α. Conclusión. El TNF-α fue el principal responsable de la inhibición de la proliferación en las células 32Dcl3 tratadas con caseinato de sodio o caseína α.

Abstract Introduction: Sodium caseinate (CS) and its components (alpha-casein, beta-casein, and kappa-casein) have been shown to inhibit the proliferation of the mouse hematopoietic 32D clone 3 (32Dcl3) cell line and induce its differentiation into macrophages. It is well-known that alpha-casein induces IL-1β production and that this cytokine inhibits the proliferation via the production of tumor necrosis factor alpha (TNF-alpha), but it is not known if CS and the caseins inhibit the proliferation via TNF-alpha production. Objective: To evaluate if CS and alpha-casein, beta-casein and kappa-casein inhibit the proliferation on 32Dcl3 cell line via TNF-alpha. Materials and methods: We used different concentrations of CS, alpha-casein, beta-casein and kappa-casein in 32Dcl3 cells to evaluate cell proliferation. We assessed cell viability by MTT, induction to apoptosis by flow cytometry, and TNF-alpha synthesis by ELISA. Additionally, we performed anti-TNF-alpha neutralization assays on 32Dcl3 cells treated with CS and alpha-casein and we evaluated proliferation. Results: The results showed that CS, alpha-casein, beta-casein, and kappa-casein reduced proliferation of the 32Dcl3 cell line without affecting the viability and that only CS and alpha-casein induced apoptosis and the release of TNF-alpha. The 32Dcl3 cells treated with CS and alpha-casein reestablished their proliferation by using anti-TNF-alpha antibodies. Conclusion: TNF-alpha was the main responsible for the inhibition of proliferation in 32Dcl3 cells treated with CS or alpha-casein.

Caseina em sistemas de produção de São Luis de Montes Belos, GO / Caseine in production systems in the São Luis de Montes Belos, GO

A caseína é encontrada no leite de vaca, perfazendo cerca de 80% do conteúdo proteico desse alimento, possuindo alto valor econômico para a indústria láctea. Objetivou-se avaliar o teor de caseína no leite de propriedades rurais que pertencem ao Arranjo Produtivo Lácteo (APL) de São Luís de Montes Belos, Goiás. Coletaram-se 158 amostras de leite de tanques de expansão de propriedades da região do APL. A determinação da composição de caseína foi realizada pelo método de infravermelho. Com os resultados obtidos foi determinada a frequência absoluta e relativa de amostras com teor de caseína consideradas em acordo ou desacordo com o preconizado pela literatura. Observou-se que 35,45% das amostras coletadas estavam em desacordo. Conclui-se que há necessidade da adoção de medidas relacionadas ao manejo nutricional e ou sanitário dos rebanhos para que se consiga um leite com qualidade.

An in Vitro Evaluation of Remineralizing Capacity of Self-Assembling Peptide (SAP) P11-4 and Casein Phosphopeptides-Amorphous Calcium Phosphate (CPP-ACP) on Artificial Enamel

Objective: To determine and compare the remineralizing capacity of self-assembling peptide (SAP) P11-4 and casein phosphopeptides­amorphous calcium phosphate (CPP-ACP) on enamel. Material and Methods: Enamel samples were divided into 2 groups. Group I was treated with Self­assembling peptide (SAP) P11-4 and group II with casein phosphopeptides-amorphous calcium phosphate (CPP-ACP). In both groups, remineralizing capacity was assessed at baseline, 2 weeks, 6 weeks and 12 weeks. Student's t- test and ANOVA were applied, with the significance level set at 5%. Results: The mean calcium weight % was evaluated at baseline, 2 weeks, 6 weeks and 12 weeks. In Group I, there was increase in mean value (62.12 ± 1.24) from baseline to 12 weeks (67.36 ± 2.14). However, there was decrease in phosphate weight % from 37.16 ± 2.52 at baseline to 35.72 ± 2.11 at 12 weeks. In Group II, mean calcium weight % was 64.18 ± 1.52 at baseline, which ultimately increased to 66.01 ± 2.03 at 12 weeks. Phosphate weight % showed reduction from 37.34 ± 2.23 at baseline to 35.04 ± 2.02 at 12 weeks. Ca/P ratio showed significant improvement. There was significant difference in Ca/P ratio at 2 weeks, 6 weeks and 12 weeks in both groups (p<0.05). Conclusion: Self-assembling peptide (SAP) P11-4 found to be more effective and efficient as compared to casein phosphopeptides­amorphous calcium phosphate (CPP-ACP).

Comparative Evaluation of Remineralizing Potential of a Paste Containing Bioactive Glass and a Topical Cream Containing Casein Phosphopeptide-Amorphous Calcium Phosphate: An in Vitro Study

Objective: To evaluate and compare the remineralization potential of a dentifrice containing bioactive glass and a topical cream containing casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) in remineralizing artificial carious lesion on enamel. Material and Methods: Forty-five freshly extracted human permanent premolar teeth were selected. Samples were divided into three groups: GI - regular tooth paste without specific remineralizing agent; GII - tooth paste containing calcium sodium-phosphosilicate (novamin) and GIII - topical cream containing casein phosphopeptide-amorphous calcium phosphate. All the sound enamel samples were viewed under scanning electron microscope (SEM) to assess the topographical pictures of enamel surface and energy dispersing x-ray analysis (EDAX) was done to estimate quantitatively the amounts of mineral (calcium and phosphorous). The mineral content of calcium and phosphorus after demineralization in each group was noted. The samples were then subjected to SEM and EDAX. Results: GI does not show any increase in the calcium and phosphorus after applying toothpaste without any remineralizing agent but GII and GIII showed a net increase in calcium and phosphorous values after applying concern-remineralizing agents. Inter group comparison showed GIII yield higher net calcium and phosphorous values than GII. Conclusion: Two remineralizing agents showed remineralization potential on enamel surfaces. Casein phosphopeptide-amorphous calcium phosphate showed better remineralizing potential than calcium sodium phosphosilicate. Hence CPP-ACP can be considered as the material of choice in remineralizing early enamel carious lesions.

The in vitro remineralizing effect of CPP-ACP and CPP-ACPF after 6 and 12 weeks on initial caries lesion

Abstract Objective: The aim of this in vitro study was to determine the effects of remineralization promoting agents containing casein phosphopeptide-stabilized amorphous calcium phosphate (CPP-ACP), or CPP-ACP in combination with fluoride (CPP-ACPF) on artificial white spot lesions (WSLs) after 6 and 12 weeks. Methodology: White spot lesions were created on 123 sectioned premolars (246 specimens) with a demineralization solution during a 96 hours pH-cycling regime. Two experimental groups were created: a CPP-ACP group (Tooth Mousse™), and a CPP-ACPF group (Mi Paste Plus™). Additionally, two control groups were created, one using only a conventional toothpaste (1450 ppm fluoride) and another one without any working agents. All teeth were also daily brushed with the conventional toothpaste except the second control group. Tooth Mousse™ and Mi Paste Plus™ were applied for 180 seconds every day. The volume of demineralization was measured with transverse microradiography. Six lesion characteristics regarding the lesion depth and mineral content of WSLs were also determined. Results: The application of CPP-ACP and CPP-ACPF had a significant regenerative effect on the WSLs. Compared to Control group 1 and 2 the volume of demineralization after 6 weeks decreased significantly for CPP-ACP (respectively p<0.001 and p<0.001) and CPP-ACPF (respectively p=0.001 and p=0.003). The same trend was observed after 12 weeks. For the CPP-ACPF group, WSL dimensions decreased significantly between 6 and 12 weeks follow-up (p=0.012). The lesion depth reduced significantly after application of CPP-ACP and CPP-ACPF but increased significantly in the Control groups. Mineral content increased for CPP-ACP and CPP-ACPF after an application period of 12 weeks, but this was only significant for CPP-ACP. Conclusions: Long-term use of CPP-ACP and CPP-ACPF in combination with a conventional tooth paste shows beneficial effects in the recovery of in vitro subsurface caries lesions.

Effects of in-office bleaching agent combined with different desensitizing agents on enamel

Abstract Objective: To analyze color change, microhardness and chemical composition of enamel bleached with in-office bleaching agent with different desensitizing application protocols. Materials and Methods: One hundred and seventeen polished anterior human enamel surfaces were obtained and randomly divided into nine groups (n = 13). After recording initial color, microhardness and chemical composition, the bleaching treatments were performed as G1: Signal Professional White Now POWDER&LIQUID FAST 38% Hydrogen peroxide(S); G2: S+Flor Opal/0.5% fluoride ion(F); G3: S+GC Tooth Mousse/Casein Phosphopeptide-Amorphous Calcium Phosphate (CPP-ACP) paste(TM); G4: S+UltraEZ/3% potassium nitrate&0.11% fluoride(U); G5: S+Signal Professional SENSITIVE PHASE 1/30% Nano-Hydroxyapatite (n-HAP) suspension(SP); G6: S-F mixture; G7: S-TM mixture; G8: S-U mixture; G9: S-SP mixture. Color, microhardness and chemical composition measurements were repeated after 1 and 14 days. The percentage of microhardness loss (PML) was calculated 1 and 14 days after bleaching. Data were analyzed with ANOVA, Welch ANOVA, Tukey and Dunnett T3 tests (p<0.05). Results: Color change was observed in all groups. The highest ΔE was observed at G7 after 1 day, and ΔE at G8 was the highest after 14 days (p<0.05). A decrease in microhardness was observed in all groups except G6 and G7 after 1 day. The microhardness of all groups increased after 14 days in comparison with 1 day after bleaching (p>0.05). PML was observed in all groups except G6 and G7 after bleaching and none of the groups showed PML after 14 days. No significant changes were observed after bleaching at Ca and P levels and Ca/P ratios at 1 or 14 days after bleaching (p>0.05). F mass increased only in G2 and G6, 1 day after bleaching (p<0.05). Conclusions: The use of desensitizing agents containing fluoride, CPP-ACP, potassium nitrate or n-HAP after in-office bleaching or mixed in bleaching agent did not inhibit the bleaching effect. However, they all recovered microhardness of enamel 14 days after in-office bleaching.

A Case of an Infant Diagnosed with Cow's Milk Allergy and Concurrent Meat Allergy via ImmunoCAP ISAC® / 고신대학교의과대학학술지

Approximately 13–20% of infants with milk allergies concurrently exhibit beef allergies. Here, we report a 24-month-old infant who exhibited both pork and beef allergies, concurrently with a milk allergy. The infant's laboratory test results were: 3.73 ISU-E (ISAC standardized unit for IgE) for cow milk β-lactoglobulin, 23.8 ISU-E for casein, 12.8 ISU-E for cow milk Bos d 6 of serum albumin, and 4.85 ISU-E for cat Fel d 2. This case report summarizes an infant patient diagnosed with a meat allergy that was associated with cow's milk allergy, using ImmunoCAP ISAC®. Not only ImmunoCAP ISAC® but also immunocap can be used to diagnose milk allergy and meat allergy at the same time, immunocap testing for component antigen is rare. ImmunoCAP ISAC® is used to diagnose these allergies in our case study, as it has advantage that only 1ml of blood is needed to run various component antigen tests.