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.

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.

In Situ Effect of Intra-Oral Application of Pastes Containing CPP-ACP or CPP-ACPF Against Initial Enamel Erosion

Objective: To evaluate in situ the effect of toothpastes containing casein phosphopeptide - amorphous calcium phosphate (CPP-ACP) and casein phosphopeptide-amorphous calcium phosphate associated to fluoride (CPP-ACPF) on initial erosion prevention. Material and Methods: Bovine enamel blocks (n = 192) were randomly assigned into 4 phases according to the baseline surface hardness: GI: CPP-ACP Paste (MI Paste™), GII: CPP-ACPF Paste (MI Paste Plus™), GIII: Fluoridated paste and GIV: Placebo Paste. In each of the 4 crossover phases, twelve volunteers wore intraoral palatal appliances containing 4 enamel blocks for 2 hours, then the tested treatments were applied intraorally (3 min) and the appliance was maintained in the mouth for another 3 hours. After, the appliances were removed and immersed in hydrochloric acid (0.01 M, pH 2.3) for 30 seconds to promote erosive demineralization. The final surface hardness was evaluated and percentage of surface hardness loss was calculated. The data were analyzed by ANOVA and Tukey's test (α = 5%). Results: The application of CPP-ACP paste, independent of fluoride content, resulted in significant lower enamel hardness loss (GI: 9.26% ±3.48 and GII: 9.14% ±1.73) compared to NaF (GIII: 15.5% ± 3.94) and placebo (GIV: 16.7% ± 4.07) pastes, which did not show difference between them. Conclusion: The CPP-ACP pastes were able to reduce initial erosive demineralization in relation to fluoride and placebo pastes. Nevertheless the formulation of CPP-ACP with fluoride did not provide an additional benefit.

Association of casein micelle size and enzymatic curd strength and dry matter curd yield / Associação do tamanho das micelas de caseína com a força e produção de massa da coalhada

ABSTRACT: The aim of the present study was to explore the association between milk protein content and casein micelle size and to examine the effects of casein micelle size on enzymatic curd strength and dry matter curd yield using reduced laboratory-scale cheese production. In this research, 140 bulk tank milk samples were collected at dairy farms. The traits were analyzed using two linear models, including only fixed effects. Smaller micelles were associated with higher κ-casein and lower αs-casein contents. The casein micellar size (in the absence of the αs-casein and κ-casein effects) did not affect the enzymatic curd strength; however, smaller casein micelles combined with higher fat, lactose, casein and κ-casein contents exhibited a favorable effect on the dry matter curd yield. Overall, results of the present study provide new insights into the importance of casein micelle size for optimizing cheese production.

RESUMO: Este trabalho foi desenvolvido com o objetivo de investigar a associação da composição proteica do leite com o tamanho das micelas de caseína, e o efeito do TMCN sobre a firmeza do coágulo enzimático e da produção de massa seca do coágulo produzido em escala reduzida. Foram coletadas 140 amostras de leite cru de diferentes fazendas. Os dados foram analisados usando dois modelos lineares, incluindo somente efeitos fixos. Menores micelas de caseína foram associadas com maior conteúdo de k-caseína e menor conteúdo de αs-caseína. O tamanho das micelas de caseína sem o efeito da αs-caseína e k-caseína não apresentou efeito sobre a firmeza do coágulo, porém apresentou efeito significatico sobre a produção de massa seca do coágulo. Esses resultados demonstram a importância do tamanho das micelas de caseína para otimizar a produção de queijo.

CPP-ACP complexe as an alternative to treatment of incisor molar hypomineralization: case report / Complexo CPP-ACP como alternativa ao tratamento da hipomineralização molar incisivo: relato de caso

ABSTRACT The objective of this study was to report the clinical treatment of a child with Incisor Molar Hipomineralization. A 5-year-old Brazilian child, male gender, was diagnosed with Molar Incisor Hypomineralization, reporting high teeth sensitivity. After anamnesis and clinical examination, treatment was conducted with three weekly applications of fluoride varnish containing 5% CPP-ACP complex. Also, it was advice to the patient for using a toothpaste containing fluoride and CPP-ACP (MI Paste Plus). After that, molars with great tooth structure loss were restored with resin modified glass ionomer cement. Prior to the first topical application of varnish with CPP-ACP and fluoride toothpaste containing CPP-ACP, a sensitivity test was conducted using thermal stimulus and facial pain scale. It was observed relative sensitivity decrease between sessions, reporting no sensitivity at the last session before the restoration. The treatment of Molar Incisor Hypomineralization teeth with CPP-ACP complex associated with fluoride varnish can be an alternative to reduce sensivity.

RESUMO O objetivo desse estudo foi relatar o tratamento clínico de uma criança com Hipomineralização Molar Incisivo e, portanto, alto índice de sensibilidade dentária. Uma criança de cinco anos de idade, do gênero masculino, foi diagnosticada com Hipomineralização Molar Incisivo, relatando alta sensibilidade dentária. Após a anamnese e exame clínico, o tratamento iniciou-se com três aplicações de verniz fluoretado a 5% com complexo de CPP-ACP, associado à um dentifrício fluoretado contendo CPP-ACP (MI Paste Plus), em intervalos semanais. Após as três sessões de aplicação de verniz, os molares com grande perda de estrutura dentária foram restaurados com cimento de ionômero de vidro modificado por resina (Vitremer). Previamente à primeira aplicação tópica do verniz fluoretado com CPP-ACP e utilização do dentifrício fluoretado contendo CPP-ACP foi realizado um teste de sensibilidade dentária usando estímulo térmico e escala facial de dor, repetindo-se a cada sessão. Verificou-se que a sensibilidade diminuiu entre as sessões, com ausência de sensibilidade na última sessão antes da restauração. O tratamento de dentes com Hipomineralização Molar Incisivo utilizando-se o complexo CPP-ACP associado à aplicação tópica de flúor pode ser uma alternativa para redução da sensibilidade dentária.

Influence of biofilm formation on the mechanical properties of enamel after treatment with CPP-ACP crème

Abstract: The study aimed to investigate the effects of bacterial biofilms on changes in the surface microhardness of enamel treated with casein phosphopeptide—amorphous calcium phosphate (CPP-ACP) with and without fluoride. Human enamel blocks with incipient caries-like lesions were divided into four groups of 13: G1: Saliva (Control); G2: fluoride dentifrice (Crest™, 1100 ppm as NaF); G3: CPP-ACP (MI Paste; Recaldent™); and G4: CPP-ACPF (MI Paste Plus; Recaldent™ 900 ppm as NaF). The specimens were soaked in demineralizing solution for 6 h and remineralized in artificial saliva for 18 h alternately for 10 days. The dentifrice was prepared with deionized water in a 1 : 3 ratio (w/w) or applied undiluted in the case of the CPP-ACP group. The surface microhardness (SMH) was evaluated at baseline, after artificial caries, after pH cycling and treatment with dentifrices, and after incubation in media with Streptococcus mutans for biofilm formation. The biofilms were exposed once a day to 2% sucrose and the biofilm viability was measured by MTT reduction. The percentage of change in surface microhardness (%SMHC) was calculated for each block. The data were analyzed by nonparametric test comparisons (α = 0.05). The %SMHC values observed in G2 were different from those of G1, G3, and G4 (p < 0.05). After biofilm formation, %SMHC was positive in G2 and G4 when compared to G1 and G3, but resistance to demineralization after biofilm formation was similar in all groups. In conclusion, the presence of biofilms did not influence the treatment outcomes of anticaries products.

Produtos lácteos podem contribuir com a prevenção da cárie dentária? Scoping review / Can milk and dairy products help with dental caries prevention? Scoping review

Objetivo: A associação entre leite e cárie dentária ainda é objeto de debate. Nesta scoping review, verificou-se a relação entre ingestão de leite e laticínios e a prevenção da cárie dentária. Material e método: Foi realizada pesquisa bibliográfica, a partir de consulta a artigos indexados nas bases de dados PubMed, Scielo e Lilacs publicados entre 2006 e janeiro/2016, incluindo estudos clínicos e laboratoriais, publicados nas línguas inglês, português e espanhol. Resultados: Dentre as 96 publicações inicialmente identificadas, apenas seis contemplaram os critérios de inclusão e exclusão. Os artigos apresentaram metodologias diversas que investigaram o efeito do leite, caseína, proteína do soro e compostos à base de fosfopeptídeos da caseína e fosfato de cálcio amorfo. O efeito preventivo dessas substâncias foi mais observado em crianças e em amostras de dentes extraídos. Conclusão: A diversidade metodológica dificultou comparações e conclusões, mas apontou para o potencial do uso do leite e laticínios no controle da cárie.

Aim: The association between milk and dental caries remains controversial. In this scoping review, we checked the relation between intake of milk and dairy products with the prevention of dental caries. Material and Methods: The literature search followed the protocol based on papers indexed in the databases PubMed, Scielo and Lilacs, published between 2006 and January/2016, including clinical studies and laboratories, published in English, Portuguese and Spanish. Results: Among the 96 publications initially identified, only six contemplated the inclusion and exclusion criteria. The papers showed diversities of methodologies that investigate the effect of milk, casein, whey protein and protein compounds of casein phosphopeotidies and amorphous calcium phosphate. The preventive effect of those substances was more observed in children and in samples of extracted teeth. Conclusion: The methodological diversity of studies hampers the comparisons and conclusions, but pointed out to a potential use of milk and dairy products in the control of dental caries.

Effect of Chewing Gum Containing CPP-ACP on Salivary Flow and Buffer Capacity: An in vivo Study

Objective: To evaluate salivary flow and buffer capacity by means of mechanical and chemical-mechanical stimuli, through the use of chewing gums. Material and Methods: The study was a cross-sectional study with 12 volunteers, divided into three groups, in three phases: Group A: paraffin gum; Group B: Chewing gum without sucrose, flavored (Trident®); Group C: Flavored chewing gum, without sucrose and amorphous calcium casein-phosphate phosphopeptide (Trident Total®). The stimulated total saliva was collected after 5 minutes of mastication of one of the products and the volume was expressed in mL / min. The same sample was submitted to pH measurement with the use of a digital potentiometer, where the results were classified in normal buffer capacity (final pH between 5.0 and 7.0) or low (final pH <4.0). The results were evaluated regarding the normality of the sample distribution (Shapiro-Wilk test), Analysis of Variance (ANOVA) and Tukey's test. Results: Chewing gums increased the salivary flow of the volunteers, when compared to the control group (paraffin) (1.53 mL / min), differing statistically from the group, although there was no difference between Trident® (2.09 mL / Min) and Trident Total® (2.06mL / min). Regarding the buffer capacity, the values obtained were 6.94 (paraffin), 6.99 (Trident®) and 6.93 (Trident Total®), with no difference between groups (p = 0.713). Conclusion: It was concluded that chewing gums, with and without CPP-ACP, increased the salivary flow in relation to the control group. In relation to buffer capacity the values obtained for chewing gums with and without CPP-ACP, are shown to be within the normal range.

Effects of glycomacropeptide in human milk and formula milk on proliferation ofbiifdobacterium infantis in vitro / 中华围产医学杂志

ObjectiveTo explore the effects of glycomacropeptide (GMP) in human milk and formula milk on proliferation ofbifidobacterium infantis and their dose-response relationship.Methods Casein was isolated from the milk of 30 healthy postpartum women from Guangzhou Women and Children's Medical Center in September 2014, and hydrolyzed by rennet to obtain GMP, which was then purified by ultrafiltration and ion exchange chromatography. Human milk GMP and cow milk GMP (0, 250, 500, 1 000, 1 500, 2 000 and 3 000 mg/L) were added tobifidobacterium infantis liquid medium, and cultured under anaerobic conditions. Concentration of bacteria was measured by turbidimetric microplate assay (detection of OD600 nmvalue of medium). Difference of proliferative activities ofbiifdobacterium infantis in human milk GMP and cow milk GMP was compared with independent samplest-test.ResultsPurified human milk GMP concentration was 1 712.20 mg/L, with a purity of 80.3%. Increasing the cow milk GMP initial concentration in the culture medium at 250-2 000 mg/L could increase the concentration and proliferation rate ofbiifdobacteria infantis. When cultured at 36 h with GMP of various concentrations, the proliferation ofbiifdobacteria infantis maintained at a logarithmic phase. Therefore, 36 h was chosen as the test time point to compare the proliferation ofbifidobacterium infantis. At 36 h, when GMP in the medium was 1 000, 1 500, 2 000 and 3 000 mg/L, concentrations ofbiifdobacteria infantis in human milk GMP were 2.255±0.036, 2.583±0.088, 2.877±0.080 and 3.219±0.081, respectively, which were significantly higher than those in cow milk GMP (2.115±0.053, 2.312±0.064, 2.542±0.090 and 2.894±0.076;t=4.867, 5.569, 6.192 and 6.516; allP<0.01).Conclusions Both human milk GMP and cow milk GMP can promote the proliferation ofbiifdobacterium infantisin vitro, and the proliferative activity in human milk is greater than in cow milk at the same concentration of GMP.

Hydrolyzed collagen (gelatin) decreases food efficiency and the bioavailability of high-quality protein in rats / Colágeno hidrolisado (gelatina) reduz a eficiência alimentar e a biodisponibilidade de proteína de alta qualidade em ratos

Objective Although deficient in all indispensable amino acids, gelatin is used in protein-restricted diets. Food efficiency and protein quality of casein and gelatin mixtures in low protein diets in Wistar rats were investigated. Methods The rats were treated with protein-restricted diets (10.0 and 12.5%) containing casein (control diets), casein with gelatin mixtures (4:1 of protein content), and gelatin as sources of protein. The food conversion ratio, protein efficiency ratio, relative and corrected protein efficiency ratio, true protein digestibility, and hepatic parameters were estimated. Results After 28 days of the experiment, food efficiency of 10.0% casein/gelatin diet decreased when compared to that of 10.0% casein diet, and the protein efficiency ratio of the casein/gelatin mixtures (10.0%=2.41 and 12.5%=2.03) were lower than those of the casein (10.0%=2.90 and 12.5%=2.32). After 42 days of the experiment, the weight of the liver of the animals treated with 10.0 and 12.5% casein/gelatin diets, and the liver protein retention of the 12.5% casein/gelatin diet group of animals were lower than those of the control group. Conclusion Gelatin decreases food efficiency and high-quality protein bioavailability in protein-restricted diets. .

Objetivo A gelatina é deficiente em todos os aminoácidos indispensáveis, mas é usada em dietas com restrição de proteína. A eficiência alimentar e a qualidade da proteína de misturas de caseína com gelatina em dietas com baixo teor de proteína foram investigadas em ratos Wistar. Métodos Ratos foram tratados com dietas restritas em proteína (10,0 e 12,5%), contendo, como fonte de proteína: caseína (dieta controle), misturas de caseína com gelatina (4:1 do teor de proteína) e gelatina. Foram estimados os seguintes índices: taxa de conversão alimentar, quociente de eficiência proteica, quociente de eficiência proteica relativo e corrigido, digestibilidade verdadeira da proteína e parâmetros hepáticos. Resultados Após 28 dias de experimento, a eficiência alimentar da dieta caseína:gelatina a 10,0% diminuiu em comparação com a dieta de caseína a 10,0%, e o quociente de eficiência proteica das misturas caseína: gelatina (10,0%=2,41 e 12,5%=2,03) foi menor do que aqueles da caseína (10,0%=2,90 e 12,5%=2,32). Após 42 dias de experimento, o peso do fígado dos animais tratados com mistura caseína:gelatina a 10,0 e 12,5% e a retenção proteica no fígado dos animais do grupo caseína: gelatina a 12,5% diminuíram em comparação ao grupo-controle. Conclusão Gelatina reduz a eficiência alimentar e a biodisponibilidade de proteína de alta qualidade em dietas restritas em proteínas. .

The effect of CPP-ACP and Nd:YAG laser on the bond strength of softened dentin

The purpose of this study was to investigate the effect of CPP-ACP treatment and Nd:YAG laser on microtensile bond strength (µTBS) of softened dentin. Sixty samples were obtained from thirty sound third molars. All samples were submitted to dentin softening procedure, by the immersion of the specimens in 30 mL of Sprite Zero for 30min. Afterwards, the samples were randomly divided according to the CPP-ACP treatment: CG-Control group; MP-treated with CPP-ACP paste (MI Paste); MPP-treated with CPP-ACP+900 ppm NaF paste (MI Paste Plus). Each group was further divided according to bonding procedure: NL-No laser; L–Laser irradiation after adhesive application and before polymerization. The laser parameters used were 1.4 W, 10 Hz, 140 mJ/pulse, with an optic fiber of 320 µm, generating energy of 174 J/cm2 per pulse. All samples were restored with Clearfil SE Bond/Filtek Z350 XT. After 24 h, the restored samples were cut into beams (± 1 mm2adhesive interface area) and subjected to a µTBS test. Data were analyzed by two-way ANOVA test and Holm-Sidak post-hoc method (α = 0.05). The treatment with CPP-ACP pastes did not significantly affect softened dentin µTBS (p = 0.070). Statistic revealed significant reduction on µTBS values for CG/L, leading to the rejection of the second null hypothesis (p < 0.001). Both CPP-ACP based pastes did not affect µTBS of softened dentin for the adhesive system utilized. The Nd:YAG laser irradiation after application of adhesive system did affect µTBS values of softened dentin samples untreated with CPP-ACP based pastes.

Usefulness of casein specific IgE and IgG antibodies to immediate type cow's milk allergy

PURPOSE: Cow's milk-specific IgE (CM-IgE) has been proposed as one of the valuable markers for diagnosis of clinical cow's milk (CM) allergy. In this study, we evaluated the additional usefulness of casein-specific IgE (casein-IgE) and IgG (casein-IgG) for the diagnosis of clinical CM allergy. METHODS: Fifty-eight subjects, aged from 3 months to 154 months, were enrolled. Thirty-four patients showed immediate-type of clinical CM allergy, and 24 patients were atopic controls. The serum levels of CM-IgE, casein-IgE, and casein-IgG were measured. Patients were divided into 2 groups: those aged under 12 months and those aged 12 months or over. The diagnostic values of each antibody were analyzed and compared using the Mann-Whitney U-test and receiver operating characteristic curves. RESULTS: CM allergy had significantly higher levels of CM-IgE and casein-IgE, and lower levels of casein-IgG/IgE ratio when compared to atopic controls in both age groups (P<0.05). CM-IgE and casein-IgE were shown to be better predictive markers for immediate-type CM allergy in patients under 12 months, while casein-IgG/IgE ratio was a more useful marker in those aged 12 months or over. Considering 100% positive predictive values, cutoff points were 1.04 kU/L for CM-IgE, 0.11 kU/L for casein-IgE, 19.5 for casein-IgG/IgE ratio in patients aged under 12 months, and 7.1 kU/L for CM-IgE, 1.41 kU/L for casein-IgE, 12.51 for casein-IgG/IgE ratio in those aged 12 months or over. CONCLUSION: CM-IgE, casein-IgE, and casein-IgG/IgE ratio are useful markers for predicting immediate-type CM allergy. Further studies are needed on diagnostic decision points for CM allergy using combination of cutoff values of these 3 markers.

Sodium fluoride and casein phosphopeptide-amorphous calcium phosphate cream plus sodium fluoride efficacy in preventing enamel erosion in a simulated oral environment study model.

Aim: The aim of this study was to evaluate the effectiveness of dentifrices containing high concentrations of sodium fluoride (NaF) and casein phosphopeptide‑amorphous calcium phosphate cream plus fluoride (CPP‑ACPF) in prevention of the erosion in a simulated oral environment study model. Subjects and Methods: Fifteen flat human enamel specimens were polished and half of the surfaces were protected with adhesive tape. Initial Knoop microhardness (KHN) and surface roughness (SR) were measured, and specimens were assigned to four groups: Control (placebo toothpaste – G1); CPP‑ACPF (G2), NaF 1450 ppm (G3), and NaF 5000 ppm (G4). Enamel surfaces were brushed 3 times daily in association with demineralization‑remineralization cycles (5s in cola drink + 5s in artificial saliva/10 cycles/twice daily) and the specimens were maintained in a salivary flow simulator apparatus. After 14 days, KHN and SR were measured again, and the enamel surfaces were analyzed by scanning electronic microscopy (SEM). Statistical Analysis Used: Data were analyzed using the two‑way ANOVA and Student–Newman– Keuls multiple range test (α =0.05). Results: All the tested groups presented a decrease in KHN after 14 days (P < 0.05). There was no statistical significance among materials tested. Significant increase in SR was observed for all groups. SEM analysis showed morphological alterations with honeycomb structures in enamel surfaces in the four experimental groups. Conclusions: It was concluded that tooth brushing with dentifrices with high concentration of NaF and CPP‑ACPF cream was not able to prevent enamel erosion in simulated oral environment.

A Milk Protein, Casein, as a Proliferation Promoting Factor in Prostate Cancer Cells

PURPOSE: Despite most epidemiologic studies reporting that an increase in milk intake affects the growth of prostate cancer, the results of experimental studies are not consistent. In this study, we investigated the proliferation of prostate cancer cells treated with casein, the main protein in milk. MATERIALS AND METHODS: Prostate cancer cells (LNCaP and PC3), lung cancer cells (A459), stomach cancer cells (SNU484), breast cancer cells (MCF7), immortalized human embryonic kidney cells (HEK293), and immortalized normal prostate cells (RWPE1) were treated with either 0.1 or 1 mg/mL of alpha-casein and total casein extracted from bovine milk. Treatments were carried out in serum-free media for 72 hours. The proliferation of each cell line was evaluated by an 3-(4,5-Dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. RESULTS: alpha-Casein and total casein did not affect the proliferations of RWPE1, HEK293, A459, SNU484, MCF7, HEK293, or RWPE1 cells. However, PC3 cells treated with 1 mg/mL of alpha-casein and casein showed increased proliferation (228% and 166%, respectively), and the proliferation of LNCaP cells was also enhanced by 134% and 142%, respectively. The proliferation mechanism of alpha-casein in PC3 and LNCaP cells did not appear to be related to the induction of Insulin-like growth factor-1 (IGF-1), since the level of IGF-1 did not change upon the supplementation of casein. CONCLUSIONS: The milk protein, casein, promotes the proliferation of prostate cancer cells such as PC3 and LNCaP.

A Milk Protein, Casein, as a Proliferation Promoting Factor in Prostate Cancer Cells

PURPOSE: Despite most epidemiologic studies reporting that an increase in milk intake affects the growth of prostate cancer, the results of experimental studies are not consistent. In this study, we investigated the proliferation of prostate cancer cells treated with casein, the main protein in milk. MATERIALS AND METHODS: Prostate cancer cells (LNCaP and PC3), lung cancer cells (A459), stomach cancer cells (SNU484), breast cancer cells (MCF7), immortalized human embryonic kidney cells (HEK293), and immortalized normal prostate cells (RWPE1) were treated with either 0.1 or 1 mg/mL of alpha-casein and total casein extracted from bovine milk. Treatments were carried out in serum-free media for 72 hours. The proliferation of each cell line was evaluated by an 3-(4,5-Dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. RESULTS: alpha-Casein and total casein did not affect the proliferations of RWPE1, HEK293, A459, SNU484, MCF7, HEK293, or RWPE1 cells. However, PC3 cells treated with 1 mg/mL of alpha-casein and casein showed increased proliferation (228% and 166%, respectively), and the proliferation of LNCaP cells was also enhanced by 134% and 142%, respectively. The proliferation mechanism of alpha-casein in PC3 and LNCaP cells did not appear to be related to the induction of Insulin-like growth factor-1 (IGF-1), since the level of IGF-1 did not change upon the supplementation of casein. CONCLUSIONS: The milk protein, casein, promotes the proliferation of prostate cancer cells such as PC3 and LNCaP.

Polimorfismos genéticos da kappa-caseína e da beta-lactoglobulina e produção de leite em bovinos / Kappa-casein and beta-lactoglobulin genetic polymorphisms and milk yield in bovines

As variantes gênicas da beta-lactoglobulina (β-LG) e da kappa-caseína (κ-CN) bovinas são associadas à produção, qualidade e características de processamento do leite. O objetivo deste trabalho foi analisar as frequências dos genótipos AA, AB e BB, por meio da técnica de PCR-RFLP, da β-LG e da κ-CN bovinas, e suas associações com a produção de leite (kg leite/dia) em bovinos das raças Girolanda, Holandesa e Jersey. Para a κ-CN, a frequência do genótipo AA foi maior nos animais das raças Holandesa (37%) e Girolanda (63%). Na raça Jersey, houve predomínio do genótipo BB (60%). Para a β-LG, o genótipo AB foi o mais encontrado nas raças Girolanda (54%) e Holandesa (58%), enquanto nos animais da raça Jersey houve predomínio do genótipo BB (45%). Houve associação do alelo B da κ-CN com maior produtividade leiteira nas raças Girolanda e Holandesa, e do alelo A da β-LG com maior produtividade de leite na raça Jersey. As variantes genéticas da κ-CN podem ser usadas como marcadores na seleção para a produtividade leiteira nas raças Girolanda e Holandesa. Para a raça Jersey, as variantes da β-LG seriam mais adequadas para essa seleção.

Bovine beta-lactoglobulin (β-LG) and kappa-casein (κ-CN) genic variants are associated with productivity, quality and processing features of milk. The objective of this study was to analyze through the PCR-RFLP technique, the frequency of AA, AB and BB genotypes of bovine β-LG and κ-CN, and their association to milk production (kg milk/day) in Girolanda, Holstein and Jersey cattle. For κ-CN, the frequency of the AA genotype was higher in Holstein (37%) and Girolanda (63%), while there was a predominance of the BB genotype in Jersey (60%). For β-LG, the BB genotype was the most found in Girolanda (54%) and Holstein (58%), while there was a predominance of the BB genotype (45%) in Jersey. There was a positive association between B allele of κ-CN and milk production in the Girolanda and Holstein cattle and between A allele of β-LG and milk production in the Jersey cattle. Genetic variants of κ-CN could be used as markers for the selection for productivity in Girolanda and Holstein cattle. The genetic variants of β-LG would be more appropriate for this selection in the Jersey breed.

Effects of casein on iodine metabolism in body and thyroid of mice under excessive iodine / 中国地方病学杂志

Objective To observe the effects of casein on iodine metabolism in blood,urine and thyroid of mice under excessive iodine.Methods A 2 by 3 factorial design was used in the experiment.The levels of iodine and casein were 50 and 600 μg/L in drinking water and 0( Ⅰ ),10％( Ⅱ ),20％( m ) in food,respectively.After six and twelve months,iodine content in serum,thyroid gland and urine was determined by arsenic-cerium catalyzed spectrophotometry.Results In six months,the levels of serum iodine in 50 Ⅰ,50 Ⅱ,50Ⅲ,600 Ⅰ,600 Ⅱ,and 600Ⅲ groups were (85.59 ± 8.78),(64.59 ± 9.06),(72.53 ± 11.69),(110.04 ± 9.37),(81.06 ± 9.94),(86.63 ± 19.59)μg/L,respectively; the levels of iodine content in thyroid gland were (0.21 ± 0.09),(0.29 ±0.08),(0.24 ± 0.05),(0.50 ± 0.10),(0.37 ± 0.13),(0.42 ± 0.12)g/kg,respectively; the levels of urinary iodine median were 87.5,68.1,105.5,746.5,828.3,1014.2 μg/L,respectively.The variance analysis results of factorial design showed iodine and casein significantly influenced the serum iodine level(F =27.95,18.52,all P ＜0.05),however,there was no interaction between the two(F =0.81,P ＞ 0.05); iodine significantly influenced the iodine content in thyroid gland(F =31.35,P ＜ 0.05),the presence of iodine interacted with casein(F =3.34,P ＜0.05).In twelve months,the levels of serum iodine were (88.54 ± 12.33),(72.45 ± 7.73),(72.93 ± 13.61),( 106.26 ± 12.00),(90.03 ± 7.90),( 104.88 ± 11.67)μg/L,respectively; the levels of iodine content in thyroid gland were (0.58 ± 0.12),(0.40 ± 0.14),(0.69 ± 0.16),(0.84 ± 0.13),(0.89 ± 0.13),(1.02 ± 0.11 )g/kg,respectively;the levels of urinary iodine median were 104.8,121.5,102.7,829.1,1080.8,895.2 μg/L,respectively.The variance analysis results of factorial design showed iodine and casein significantly influenced the iodine content in serum and thyroid gland (F =42.78,7.42 and 66.62,7.90,all P ＜ 0.05),however,there was no interaction between the two(F =1.93,2.31,all P ＞ 0.05).Conclusions Long-term intake of iodine can significantly increase the iodine content in serum,thyroid gland and urine,but casein may accelerate iodine excretion and partly reduce the effect.

Effects of excessive iodine and casein on histomorphalogy and ultrastructure of mouse thyroid / 中国地方病学杂志

Objective To observe the effects of casein and excessive iodine on histomorphalogY and ultrastructure of mouse thymicL Methods Based on 2 × 3 factorial design,the experimental mice were divided into 6 groupg Animal models were estabhshed by feeding the mice with different levels of iodine water and casein food.The levels of iodine were 50,600 μg/L in drinking water and 0(Ⅰ),10%(Ⅱ),20%(Ⅲ)of casein in food respectively.After 12 months,the thyroid weight was measured and the morphology of thyroid was observed under optical and electron microscope.Results Factorial analysis showed that iodine factors obviously affected the thyroid absolute and relative weiights of mice(F=16.23,9.47,P＜0.01),and there was interaction between casein and iodine(F=5.29,4.68,P＜0.01 or＜0.05).Compared wiht 150Ⅰ[(5.91±0.82)rag,(117.0±22.2)mg/kg]and 50Ⅲ[(4.90±0.63)rag,(106.1±13.3)mg/kg]groups.thyroid absolute and relative weights of the mice increased in 600 Ⅰ[(7.60±2.40)mg,(143.3±43.2)mg/kg]and 600Ⅲ[(8.63±1.88)mg,(166.2±39.4)mg/kg]groups(P＜0.05 or＜0.01),respectively.But compared with 600 Ⅰ and 600Ⅲ groups.they were reduced obviously in 600Ⅱ[(5.76±1.13)mg,(109.8±16.5)mg/kg]group(P＜0.05 or＜O.01).Colloid goiter,lymphocyte infiltration were found,some of the follicles epithelial cells appeared active under light and electron microscope in iodine excels group,which,however,decreased obviously along with the increase of casein dose.Conclusions Long-term excessive iodine may cause colloid goiter and inflammation injury of mice,possibly leading the development of thyroiditis in mice,which may be partly reduced by casein.

Kaempferol is a potent inhibitor of recombinant human protein kinase CK2 holoenzyme in vitro / 中国药理学与毒理学杂志

AIM In order to search inhibitors of protein kinase CK2, we observed the inhibitory effects of kaempferol on recombinant human protein kinase CK2 holoenzyme and its kinetics in vitro. METHODSCloning, prokaryotic expression and purification of human protein kinase CK2 α' and β subunits by gene engineering, the two subunits were mixed at equal molar ratio to reconstitute CK2 holoenzyme and identify its biological properties. The CK2 activity was assayed by detecting incorporation of 32P of [γ-32P]ATP into the substrate. The inhibitory effect of kaempferol on CK2 was assayed in the presence of different concentrations of kaempferol. Kinetic analysis of kaempferol-induced inhibition was carried out in the condition that casein concentration was fixed at 2 g·L-1 and ATP was changed at various concentrations(10, 20, 40, 80 μmol·L-1), or ATP was fixed at 10 μmol·L-1 and casein was changed at different concentrations (1, 2, 4, 8 g·L-1). RESULTS Kaempferol was shown to strongly inhibit the holoenzyme activity of recombinant human protein kinase CK2 with IC50 of 1.9 μmol·L-1, which was more effective than chrysin, morin and genistein which are both known as CK2 special inhibitors. Kinetic studies of kaempferol on recombinant human CK2 showed that kaempferol acted as a noncompetitive inhibitor with substrate ATP(Ki=1.1 μmol·L-1) and casein (Ki=3.1 μmol·L-1). CONCLUSIONKaempferol is a novel potent inhibitor of protein kinase CK2 in vitro. Discussions indicate that flavonoid inhibitors of CK2 may adopt different orientations in theactive site of CK2 and that these are determined by the number and position of their hydroxyl groups.