Effect of treatment with phosphate, casein phosphopeptide and fluoride on the remineralization: in vitro study.

This study aimed to evaluate in vitro the effect protocols and anticaries agents containing casein amorphous calcium fluoride phosphopeptide-phosphate (CPP-ACPF, MI Paste Plus), sodium trimetaphosphate (TMP) and fluoride (F), in remineralization of caries lesions. Bovine enamel blocks with initial caries lesions were divided into groups (n = 12): 1) Toothpaste without F-TMP-MI Plus (Placebo); 2) Toothpaste 1100 ppm F (1100F), 3) 1100F + MI Paste Plus (1100F-MI Paste Plus), 4) Toothpaste with 1100F + Neutral gel with 4,500 ppm F + 5%TMP (1100F + Gel TMP) and 5) Toothpaste with 1100F + Neutral gel with 9,000 ppm F (1100F + Gel F). For the 4 and 5 groups the gel was applied only once for 1 minute, initially to the study. For the 3 group, after treatment with 1100F, MI Paste Plus was applied 2x/day for 3 minute. After pH cycling, the percentage of surface hardness recovery (%SHR); integrated loss of subsurface hardness (ΔKHN); profile and depth of the subsuperficial lesion (PLM); concentrations of F, calcium (Ca) and phosphorus (P) in enamel was determined. The data were analyzed by ANOVA (1-criterion) and Student-Newman-Keuls test (p < 0.001). Treatment with 1100F alone led to ~ 28% higher remineralization when compared to treatment with 1100F associated with MI Paste Plus (p < 0.001). The 1100F and 1100F + Gel F groups showed similar values for %SHR (p = 0.150). 1100F + Gel TMP treatment also remineralized the enamel surface by ~ 30% and 20% when compared to the 1100F + Gel F and 1100F groups (p < 0.001). The lower lesion depth (ΔKHN) was observed for the 1100F + Gel TMP group (p < 0.001), where it was 54% and 44% lower in comparison to the 1100F and 1100F + Gel F groups (p < 0.001). Polarized light microscopy photomicrographs showed subsurface lesions in all groups, but these lesions were present to a lower extent in the 1100F + Gel TMP group (p < 0.001). Treatment with 1100F + Gel TMP promoted an increase in the concentration of Ca in the enamel by ~ 57% and ~ 26% when compared to the 1100F and 1100F + MI Paste Plus groups (p < 0.001), respectively. There were no significant differences between the 1100F, 1100F + MI Paste Plus and 1100F + Gel F groups (p > 0.001). Similar values of P in the enamel were observed in the 1100F, 1100F + MI Paste Plus and 1100F + Gel F groups (p > 0.001), except for the 1100F + Gel TMP group, which presented a high concentration (p < 0.001). We conclude that the 1100F+TMP gel treatment/protocol led to a significant increased remineralization when compared to the other treatments/protocols and may be a promising strategy for patients with early caries lesions.

Evaluation of the cytotoxic activity of sorafenib-loaded camel milk casein nanoparticles against hepatocarcinoma cells.

Sorafenib, a multikinase inhibitor is used to treat hepatocellular and renal carcinoma. However, a low solubility impedes its bioavailability and thus, effectiveness. This study aims to enhance its effectiveness by using novel camel milk casein nanoparticles as a delivery system. This study evaluates the cytotoxicity of sorafenib encapsulated in camel milk casein nanoparticles against human hepatocarcinoma cells (HepG2 cells) in vitro. Optimal drug loaded nanoparticles were stable for 1 month, had encapsulation efficiency of 96%, exhibited a particle size of 230 nm, zeta potential of -14.4 and poly disparity index of 0.261. Treatment with it led to cell morphology and DNA fragmentation as a characteristic of apoptosis. Flow cytometry showed G1 phase arrest of cell cycle and 26% increased apoptotic cells population upon treatment as compared to control. Sorafenib-loaded casein nanoparticles showed 6-fold increased ROS production in HepG2 cells as compared to 4-fold increase shown by the free drug. Gene and protein expression studies done by qPCR and western blotting depicted upregulation of tumor suppressor gene p53, pro-apoptotic Bax, and caspase-3 along with downregulated anti-apoptotic Bcl-2 gene and protein expression which further emphasized death by apoptosis. It is concluded regarding the feasibility of these casein nanoparticles as a delivery system with enhanced therapeutic outcomes against hepatocellular carcinoma cells.

Glutathione - IR 797 coupled Casein Nano-Trojan for augmenting the therapeutic efficacy of camptothecin in highly invasive triple negative breast cancer.

The rapid metastasis & heterogenic constitution of triple negative breast cancer (TNBC) limits drug entry to the tumor, reducing treatment effectiveness. To address this, we have synthesized Casein nanoparticles (Cn NPs) with attached glutathione (GSH), a natural ligand for cancer cell overexpressed γ-glutamyl transpeptidase (GGT). Cn NPs encapsulated with Camptothecin and NIR dye IR 797 (CCN NPs) for combinatorial therapy of TNBC. The GSH-CCN nanoparticles (CCNG NPs) act as a Nano-Trojan to deceive the cancer cells by delivering therapeutic payloads directly to specific target cells. In this study, Casein Nano-Trojan is equipped with GSH as a targeting ligand for GGT. The binding of CCNG NPs with cell surface receptors switched the anionic charge to catanionic, prompting the target cell to engulf the nanoparticles. The Casein Nano-Trojan releases its therapeutic payload inside the target cell, potentially inhibiting proliferation & inducing a high percentage of cell death (85 ± 7 %). Disintegration of mitochondrial membrane potential, inhibition of both migration & re-growth were observed. Immunofluorescence, acridine orange/ethidium bromide stain, and nuclear fragmentation assay further confirmed the substantial DNA damage induced by the high expression of γH2AX and p53. Significant therapeutic efficacy was observed in the 3D spheroids of 4T1 cells and in vivo breast cancer mice model (BALB/c). These findings demonstrate that CCNG NPs could be an effective treatment approach for highly metastatic triple negative breast cancer.

Effect of three different remineralising agents on prevention against acidic erosion of primary teeth: an in vitro study.

PURPOSE: This study aimed to evaluate and compare the protective effect of fluoride varnish (Enamelast™, Ultradent Inc., Cologne, Germany), casein phosphopeptide-amorphous calcium phosphate fluoride/CPP-ACPF (MI Paste Plus, GC Corp., Tokyo, Japan) and self-assembling P11-4 peptide (Curodont™ Protect, Credentis AG, Windisch, Switzerland), against acidic erosion of primary teeth. METHODS: Forty primary anterior teeth were randomly assigned to four groups (n = 10): group 1: control, group 2: fluoride varnish, group 3: CPP-ACPF and group 4: self-assembling P11-4 peptide. After applying remineralising agents, except for the control group, all specimens underwent an erosive challenge of carbonated soft drink and artificial saliva for 15 cycles of 6 s each at 6-h intervals for a day. Groups were compared in terms of surface microhardness, roughness readings, and surface scanning with an extra-oral scanner (D800; 3Shape A/S) before and after the erosive process. RESULTS: All experimental groups showed superior results than the control group regarding microhardness, surface roughness, and substance loss. The fluoride varnish group showed significantly favourable results in microhardness change. There was no significant difference between the experimental groups regarding surface roughness and enamel loss measurements. CONCLUSION: 5% NaF fluoride varnish, CPP-ACPF and self-assembling P11-4 peptide protect the enamel of primary teeth against erosion compared to artificial saliva alone.

An active new formulation of iron carried by aspartyl casein for iron-deficiency anemia: results of the ACCESS trial.

Oral iron supplementation is the cornerstone for the management of iron-deficiency anemia. A new oral formulation of iron conjugated with N-aspartyl-casein (Fe-ASP) (Omalin®, Uni-Pharma) is studied in the ACCESS double-blind, double-dummy randomized clinical trial; 60 patients were randomized to 12-week oral treatment twice every day either with oral ferrous sulfate (FeSO4) delivering 47 mg elementary iron or oral Fe-ASP delivering 40 mg elementary iron. Participants had hemoglobin less than 10 g/dl, decreased red blood cell (RBC) count, and ferritin lower than 30 ng/ml; patients with a medical history of malignancy were excluded. The primary endpoint was the increase of Hb in the first 4 weeks of treatment, and the study was powered for non-inferiority. A new score of global improvement was introduced where all participants were given one point for any at least 10% increase of Hb, RBC, and reticulocytes. At week 4, the mean (SE) change of Hb was 0.76 g/dl in the FeSO4 group and 0.83 g/dl in the Fe-ASP group (p: 0.876). The odds for worse allocation of the global score were 0.35 in the Fe-ASP group compared to the FeSO4 group. Patients in the Fe-ASP group experienced a significant decrease in the number of IDA-related physical signs by week 4. No differences were found between the two groups in any of the patient-reported outcomes of fatigue and of gastrointestinal adverse events either at week 4 or at week 12. ACCESS is the most recent clinical trial showing the non-inferiority of Fe-ASP to FeSO4 for the primary endpoint of the Hb change.

Diffusion-mediated carving of interior topologies of all-natural protein nanoparticles to tailor sustained drug release for effective breast cancer therapy.

Proteins are promising base materials for developing drug carriers with efficient blood circulation due to low possibilities of clearance by macrophages. However, such natural biopolymers have highly sophisticated molecular structures, preventing them from being assembled into nano-platforms with manipulable payload release profiles. Here, we report the self-assembly of two natural proteins (milk casein and rice protein) into protein nanoparticles (NPs, ∼150 nm) with tailorable release profiles. Diffusion of plant-derived paclitaxel (PTX)-containing eugenol into the hydrophobic cores of the NPs and subsequent dialysis to remove eugenol from the cores lead to the carving of the NP interiors. With the increase in the mass ratios of casein and rice protein, this process generates all-natural NPs with PTX loaded in their full cavities, semi-full cavities, or solid cores. These NPs can be efficiently uptaken by breast cancer cells and could kill the cancer cells efficiently. PTX in these NPs demonstrates increasingly sustained in vivo release profiles from full cavities, semi-full cavities, to solid cores, gradually extending its pharmacokinetic profiles in blood plasma to favor drug accumulation in breast tumor models. Consequently, the NPs with solid cores completely inhibit tumor growth in vivo, more effectively than those with full and semi-full cavities. Our work opens up a new avenue to the use of diffusion-mediated nanoscale carving in producing biomaterials with controllable interior topologies relevant to drug release profiles.

RNA sequencing and bioinformatics analysis revealed PACSIN3 as a potential novel biomarker for platinum resistance in epithelial ovarian cancer.

BACKGROUND: Failure to respond to treatment in epithelial ovarian cancer can often be attributed to platinum-based chemotherapy resistance. However, the possible mechanisms or candidate biomarkers associated with platinum resistance are yet to be elucidated, even though many researchers have performed related studies. METHODS: We performed RNA sequencing of clinical specimens obtained from patients with platinum-sensitive or resistant epithelial ovarian cancer (EOC). Furthermore, various bioinformatics approaches, including spatial analysis of functional enrichment, were used to identify key regulators and associated underlying mechanisms of platinum resistance in EOC. RESULTS: Through RNA-sequencing, we identified 263 differentially expressed genes (98 upregulated and 165 downregulated) and subjected them to Gene Oncology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses, which were characterized to the traditional platinum-resistant characteristics. Subsequently, the gene interaction network and module analysis by spatial analysis of functional enrichment software demonstrated protein kinase C and casein kinase substrate in neurons 3 (PACSIN3) as the only upregulated hub gene, and neurotensin (NTS) and KIAA0319 as downregulated hub genes in platinum-resistant EOC. We selected PACSIN3 for further analysis because it has not been studied in relation to response to platinum-based chemotherapy. PACSIN3 was significantly upregulated in ovarian cancer cells compared to immortalized human ovarian surface epithelial cells. In addition, cisplatin-induced apoptosis was measured in PACSIN3 knockout OVCA433 and BRCA-mutated EOC cell line, SNU251, by a fluorescence-activated cell sorting-based Annexin-V/propium iodide double staining assay, which revealed a significant increase in apoptosis. CONCLUSIONS: Taken together, the present study presents PACSIN3 as a promising predictive biomarker associated with platinum resistance, especially in BRCA-mutated epithelial ovarian cancers.

Pharmaceutical Potential of Casein-Derived Tripeptide Met-Lys-Pro: Improvement in Cognitive Impairments and Suppression of Inflammation in APP/PS1 Mice.

BACKGROUND: Tripeptide Met-Lys-Pro (MKP), a component of casein hydrolysates, has effective angiotensin-converting enzyme (ACE) inhibitory activity. Brain angiotensin II enzyme activates the NADPH oxidase complex via angiotensin II receptor type 1 (AT1) and enhances oxidative stress injury. ACE inhibitors improved cognitive function in Alzheimer's disease (AD) mouse models and previous clinical trials. Thus, although undetermined, MKP may be effective against pathological amyloid-ß (Aß) accumulation-induced cognitive impairment. OBJECTIVE: The current study aimed to investigate the potential of MKP as a pharmaceutical against AD by examining MKP's effect on cognitive function and molecular changes in the brain using double transgenic (APP/PS1) mice. METHODS: Experimental procedures were conducted in APP/PS1 mice (n = 38) with a C57BL/6 background. A novel object recognition test was used to evaluate recognition memory. ELISA was used to measure insoluble Aß40, Aß42, and TNF-α levels in brain tissue. Immunohistochemical analysis allowed the assessment of glial cell activation in MKP-treated APP/PS1 mice. RESULTS: The novel object recognition test revealed that MKP-treated APP/PS1 mice showed significant improvement in recognition memory. ELISA of brain tissue showed that MKP significantly reduced insoluble Aß40, Aß42, and TNF-α levels. Immunohistochemical analysis indicated the suppression of the marker for microglia and reactive astrocytes in MKP-treated APP/PS1 mice. CONCLUSION: Based on these results, we consider that MKP could ameliorate pathological Aß accumulation-induced cognitive impairment in APP/PS1 mice. Furthermore, our findings suggest that MKP potentially contributes to preventing cognitive decline in AD.

Evaluation of the changes in physical properties and mineral content of enamel exposed to radiation after treating with remineralization agent.

OBJECTIVE: The aim of this study was to investigate the effect of different remineralization agents on the physical properties and elemental content of enamel exposed to radiation. MATERIAL AND METHOD: The enamel surfaces of impacted third molar teeth were prepared, and six study groups were created (n = 6). Next, 60 Gy radiation was applied to each group. Between applications, each group except for the control group was treated with a different remineralization agent (sodium fluoride, casein phosphopeptide-amorphous calcium phosphate (CPP-ACP), casein phosphopeptide amorphous calcium phosphate with fluorite (CPP-ACFP), bioactive glass, or chitosan). The results were evaluated in terms of pre- and post-radiation values and the difference between the two. The paired-samples t test and analysis of variance test were used in the analysis of normally distributed hardness and roughness values, while Wilcoxon's signed ranks test, and the Kruskal Wallis and Mann-Whitney U tests were used in the analysis of elemental content without normal distribution. RESULTS: A statistically significant decrease was observed in microhardness measurements in all groups. Intragroup evaluation revealed a statistically significant difference between the NaF and bioactive glass groups (p < 0.05). No significant difference was observed between the groups' roughness measurements (p < 0.05). Intergroup evaluation of surface roughness revealed a significant difference in the CPP-ACFP and chitosan groups (p < 0.05). Pre- and post-radiation oxygen, magnesium, and potassium levels and Ca/P ratios also differed significantly (p < 0.05). CONCLUSION: Radiation caused a statistically significant difference in the microhardness and elemental content of enamel. However, no significant difference was observed in enamel roughness. The applied remineralizing agents have a partial ameliorating effect on the adverse impacts of radiation. CLINICAL RELEVANCE: Radiation causes changes in the mechanical properties and elemental content of tooth enamel. Remineralizing agent application is a promising option in reducing the adverse effects of irradiation.

Outcomes of dogs treated for extrahepatic congenital portosystemic shunts with thin film banding or ameroid ring constrictor.

OBJECTIVE: To compare the outcomes of dogs treated at a single institution for single extrahepatic congenital portosystemic shunts (CPSS) by thin film banding (TFB) or by placement of an ameroid constrictor (AC). STUDY DESIGN: Retrospective case series. ANIMALS: Seventy-six client-owned dogs with CPSS treated with TFB (n = 53) or AC (n = 23). METHODS: Records were reviewed for signalment, preoperative, intraoperative, and postoperative management and short-term outcomes. Data on second surgeries were reviewed. Long-term outcomes were obtained via an owner-directed health-related quality of life questionnaire. The rates of complications, mortality, and revision surgery were compared between the treatment groups. RESULTS: Postoperative complications occurred in 15 (28%) dogs with TFB (9% mortality, n = 5) and 8 (35%) dogs with an AC (4% mortality, n = 1). Long-term follow-up was available in 41 of 56 dogs at a median of 55 months (range, 15-89). Revision surgery for persistent shunting was performed in 14 (29%) dogs treated initially with TFB and in no dogs treated initially with AC (P = .007). Median long-term outcome scores were good in both groups; nine of 14 revision surgeries led to favorable outcomes. CONCLUSION: Persistent shunting requiring revision surgery was more common when CPSS were treated with TFB than with an AC, but both treatments achieved favorable long-term outcomes. CLINICAL SIGNIFICANCE: Treatment of CPPS by placement of an AC rather than TFB seems more reliable for shunt attenuation and prevention of revision surgeries.

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)

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.

White Spot Lesion Remineralisation Agents in Primary Teeth: A Systematic Review.

PURPOSE: To perform a systematic review of articles related to the clinical efficacy of topical fluoride varnishes/gels, casein phosphopeptide-amorphous calcium phosphate (CPP-ACP), and other remineralisation agents of white spot lesions (WSL) in primary teeth. MATERIALS AND METHODS: Electronic and manual searches were conducted through diverse electronic databases. The search was limited to randomised, clinical, controlled trials, and quasi-experimental papers in full-text version. Suitable individual studies were evaluated through a previously reported quality system, their data extracted and carefully analysed. RESULTS: The search identified 298 citations, and 95 were chosen and reviewed in full text. Nine relevant citations met the eligibility criteria for inclusion in the systematic review. Pair comparisons were made between fluoride varnishes, CPP-ACP, dental lasers, and control interventions. CONCLUSIONS: Clinicians are encouraged to discuss more high-quality studies in order to provide sufficient evidence and to confirm the clinical utility of remineralisation agents of WSL in primary teeth.

Avoidance of Cow's Milk-Based Formula for At-Risk Infants Does Not Reduce Development of Celiac Disease: A Randomized Controlled Trial.

BACKGROUND & AIMS: Feeding during the first months of life might affect risk for celiac disease. Individuals with celiac disease or type 1 diabetes have been reported to have high titers of antibodies against cow's milk proteins. Avoidance of cow's milk-based formula for infants with genetic susceptibility for type 1 diabetes reduced the cumulative incidence of diabetes-associated autoantibodies. We performed a randomized controlled trial in the same population to study whether weaning to an extensively hydrolyzed formula reduced the risk of celiac disease autoimmunity or celiac disease. METHODS: We performed a double-blind controlled trial of 230 infants with HLA-defined predisposition to type 1 diabetes and at least 1 family member with type 1 diabetes. The infants were randomly assigned to groups fed a casein hydrolysate formula (n = 113) or a conventional formula (control, n = 117) whenever breast milk was not available during the first 6-8 months of life. Serum samples were collected over a median time period of 10 years and analyzed for antibodies to tissue transglutaminase (anti-TG2A) using a radiobinding assay, to endomysium using an immunofluorescence assay, and antibodies to a deamidated gliadine peptide using an immunofluorometry assay. Duodenal biopsies were collected if levels of anti-TG2A exceeded 20 relative units. Cow's milk antibodies were measured during the first 2 years of life. RESULTS: Of the 189 participants analyzed for anti-TG2A, 25 (13.2%) tested positive. Of the 230 study participants observed, 10 (4.3%) were diagnosed with celiac disease. We did not find any significant differences at the cumulative incidence of anti-TG2A positivity (hazard ratio, 1.14; 95% confidence interval, 0.51-2.54) or celiac disease (hazard ratio, 4.13; 95% confidence interval, 0.81-21.02) between the casein hydrolysate and cow's milk groups. Children who developed celiac disease had increased titers of cow's milk antibodies before the appearance of anti-TG2A or celiac disease. CONCLUSIONS: In a randomized controlled trial of 230 infants with genetic risk factors for celiac disease, we did not find evidence that weaning to a diet of extensively hydrolyzed formula compared with cow's milk-based formula would decrease the risk for celiac disease later in life. Increased titers of cow's milk antibody before anti-TG2A and celiac disease indicates that subjects with celiac disease might have increased intestinal permeability in early life. ClinicalTrials.gov Number: NCT00570102.

In situ effect of CPP-ACP chewing gum upon erosive enamel loss

Abstract Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) is able to increase salivary calcium and phosphate levels at an acidic pH. Previous studies demonstrated that a CPP-ACP chewing gum was able to enhance the re-hardening of erosion lesions, but could not diminish enamel hardness loss. Therefore, there is no consensus regarding the effectiveness of CPP-ACP on dental erosion. Objective This in situ study investigated the ability of a CPP-ACP chewing gum in preventing erosive enamel loss. Material and Methods: During three experimental crossover phases (one phase per group) of seven days each, eight volunteers wore palatal devices with human enamel blocks. The groups were: GI - Sugar free chewing gum with CPP-ACP; GII - Conventional sugar free chewing gum; and GIII - No chewing gum (control). Erosive challenge was extraorally performed by immersion of the enamel blocks in cola drink (5 min, 4x/day). After each challenge, in groups CPP and No CPP, volunteers chewed one unit of the corresponding chewing gum for 30 minutes. Quantitative analysis of enamel loss was performed by profilometry (µm). Data were analyzed by Repeated-Measures ANOVA and Tukey's test (p<0.05). Results The use of chewing gum (CPP and No CPP) resulted in lower erosive enamel loss compared with the control group (p<0.05). CPP-ACP chewing gum (CPP) did not improve the protection against erosive enamel loss compared with conventional chewing gum (No CPP) (p>0.05). Conclusion The CPP-ACP chewing gum was not able to enhance the anti-erosive effect of conventional chewing gum against enamel loss.

Randomized clinical trial of the effect of NovaMin and CPP-ACPF in combination with dental bleaching

Abstract Objective This randomized, controlled, double-blind clinical study evaluated the effect of calcium sodium phosphosilicate (NovaMin) and casein phosphopeptide-amorphous calcium phosphate with fluoride (CPP-ACPF) on the prevention of post-operative sensitivity and on the effects of clinical bleaching treatment. Material and Methods Sixty volunteers were selected according to inclusion and exclusion criteria and were randomly assigned into three groups (n=20): CG (control group) patients, who were treated with 35% hydrogen peroxide; NOVAG (NovaMin group) patients, who were treated with 35% hydrogen peroxide followed by the application of NovaMin; and CPPG (CPP group) patients, who were treated with 35% hydrogen peroxide followed by the application of CPP-ACPF. Both bioactive agents were applied for five minutes. An evaporative stimulus associated with a modified visual scale was used to analyze sensitivity 24 hours after each bleaching session. The color evaluation was performed on the maxillary central incisors using a spectrophotometer. Associations between the intervention group, bleaching session, and reported sensitivity were tested using Chi-square partitioning. Results Color change values (ΔE) were analyzed using analysis of variance (ANOVA). The significance level used for both tests was 5%. In the intragroup assessment, the Friedman test showed that only the CPP-ACPF group showed no statistically significant difference (p<0.05) between baseline and first bleaching session. In the intergroup assessment, the Kruskal-Wallis test showed that the CPPG had less postoperative sensitivity after the first session, when compared to the other groups (p<0.05). Color change analysis (ΔE) showed a significant difference between the means obtained in the different bleaching sessions in all groups (p<0.05). Conclusions This study showed that the combination of CPP-ACPF with 35% hydrogen peroxide significantly reduced post-operative sensitivity in the first session, compared with the other evaluated treatments. The association of CPP-ACPF and NovaMin did not affect the color change induced by tooth bleaching.

Administration of bovine casein-derived peptide prevents cognitive decline in Alzheimer disease model mice.

There is a growing interest in identifying natural food ingredients that may serve to prevent dementia such as that due to Alzheimer disease (AD). Peptides derived from food proteins have been demonstrated to have various physiological activities such as a hypotensive action. Recent findings have indicated possible associations of hypertension with AD progression, and suggest that angiotensin converting enzyme (ACE) inhibitors with potential to pass through the blood brain barrier (BBB) may reduce the risk of AD. In this study, we investigated the effect of milk peptide (CH-3) on cognitive function in AD model mice. CH-3 contains a tripeptide (methionine-lysine-proline, MKP) that has been found to have a strong ACE inhibitory effect and the potential to pass through the BBB. Adult male ddY mice were used in this study, and an animal model of AD was induced by intracerebroventricular (ICV) injection of Aß1-42. CH-3 (250 mg/kg/day) or MKP (0.5 mg/kg/day) was orally administered every day starting 2 days before ICV injection. At 3 weeks after ICV injection, cognitive function was evaluated by the Morris water maze test. Brain samples were obtained after behavioral testing, and expression of inflammatory cytokines and NADPH oxidase subunits was measured by real-time quantitative RT-PCR. ICV injection of Aß1-42 significantly impaired cognitive function compared with that in PBS-injected mice. Daily administration of CH-3 markedly attenuated this Aß1-42-induced cognitive decline. Aß1-42 injection significantly enhanced the expression of tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS) and p22phox in the mouse hippocampus compared with PBS injection, and showed a tendency to increase the expression of monocyte chemoattractant protein-1 (MCP-1), p47phox and gp91phox, whereas CH-3 treatment markedly reduced Aß1-42-induced TNF-α, MCP-1, iNOS, p47phox and gp91phox expression. Finally, administration of MKP also attenuated Aß1-42-induced cognitive impairment with an increase in cerebral blood flow. The present study demonstrated that repeated oral administration of CH-3 to AD model mice not only improved cognitive function but also suppressed the expression of inflammatory cytokines and production of oxidative stress, and suggests its therapeutic potential for preventing cognitive impairment in AD.

Diets enriched in whey or casein improve energy balance and prevent morbidity and renal damage in salt-loaded and high-fat-fed spontaneously hypertensive stroke-prone rats.

High-fat diets induce obesity and increase risks of diabetes and cardiovascular and renal disorders. Whey- or casein-enriched diets decrease food intake and weight gain; however, their cardiovascular and renal benefits are unclear. We determined whether whey- and casein-enriched diets improve energy balance and are protective against renal damage and morbidity associated with stroke in an obesogenic and hypertensive experimental setting. We also assessed whether the hypophagic effects of these diets were due to reduced diet preference. In experiment 1, spontaneously hypertensive stroke-prone rats were randomized to (a) control (CON; 14% kcal protein, 33% fat), (b) whey (WHY; 40% protein, 33% fat), (c) casein (CAS; 40% protein, 33% fat) or (d) chow (CHW; 24% protein, 13% fat) for 12 weeks with 1% salt in drinking water for CON, WHY and CAS groups. Our results demonstrated that both WHY and CAS produced short-term hypophagia, moderately increased energy expenditure and decreased respiratory quotient, body weight and lean mass, with effects of WHY being more prolonged. Further, only WHY decreased fat mass and blood pressure. Importantly, both WHY and CAS prevented morbidity associated with stroke and decreased indices of renal inflammation (tumor necrosis factor-α, interleukin-6) and damage (osteopontin, renal lesions). In experiment 2, following four initial conditioning trials, the preference for CON, WHY or CAS diet was determined. Both WHY and CAS decreased food intake during conditioning and decreased preference. In conclusion, diets enriched in whey or casein improved energy balance, increased survival and prevented renal damage in salt-loaded and high-fat-fed spontaneously hypertensive stroke-prone rats.

Dietary Whey and Casein Differentially Affect Energy Balance, Gut Hormones, Glucose Metabolism, and Taste Preference in Diet-Induced Obese Rats.

BACKGROUND: Dietary whey and casein proteins decrease food intake and body weight and improve glycemic control; however, little is known about the underlying mechanisms. OBJECTIVE: We determined the effects of dietary whey, casein, and a combination of the 2 on energy balance, hormones, glucose metabolism, and taste preference in rats. METHODS: In Expt. 1, Obesity Prone CD (OP-CD) rats were fed a high-fat control diet (33% fat energy) for 8 wk, and then randomly assigned to 4 isocaloric dietary treatments (n = 12/group): the control treatment (CO; 14% protein energy from egg white), the whey treatment (WH; 26% whey + 14% egg white), the casein treatment (CA; 26% casein + 14% egg white), or the whey plus casein treatment (WHCA; 13% whey + 13% casein + 14% egg white) for 28 d. Measurements included food intake, energy expenditure, body composition, metabolic hormones, glucose tolerance and key tissue markers of glucose and energy metabolism. In Expt. 2, naïve OP-CD rats were randomly assigned to 3 groups (n = 8/group). During an 8 d conditioning period, each group received on alternate days either the CO or WH, CO or CA, or CO or WHCA. Subsequently, preferences for the test diets were assessed on 2 consecutive days with food intake measurements at regular intervals. RESULTS: In Expt. 1, food intake was decreased by 17-37% for the first 14 d in the WH and CA rats, and by 18-34% only for the first 4 d in the WHCA compared with the CO rats. Fat mass decreased by 21-28% for the WH rats and 17-33% for the CA rats from day 14 onward, but by 30% only on day 28 in WHCA rats, relative to CO rats. Thus, food intake, body weight, and fat mass decreased more rapidly in WH and CA rats than in WHCA rats. Energy expenditure in WH rats decreased for the first 4 d compared with CA and WHCA rats, and for the first 7 d compared with the CO rats. Circulating leptin, glucose-dependent insulinotropic polypeptide, interleukin 6, and glucose concentrations were lower in WH, CA, and WHCA rats than in CO rats. Plasma glucagon-like peptide 1 concentrations were greater in WH than in CA or WHCA rats. The improvements in glucose tolerance were greater in WH than in WHCA rats. The plasma membrane glucose transporter 4 (GLUT4)-to-total GLUT4 ratio in skeletal muscle was greater in CA and WHCA rats than in CO rats; other markers of glucose and energy metabolism in the adipose and cardiac tissues did not differ. In Expt. 2, during 4 conditioning trials, daily food intake was decreased in WH, CA, and WHCA rats by 26-37%, 30-43%, and 23-33%, respectively, compared with CO rats. Preferences for WH and CA rats were 45% and 31% lower, respectively, than those for CO rats, but that for WHCA rats did not differ. CONCLUSION: Together, these data demonstrate that in obese rats, whey, casein, and their combination improve energy balance through differential effects on food intake, taste preference, energy expenditure, glucose tolerance, and gut hormone secretion.

Short- and Long-Term Soy Diet Versus Casein Protects Liver Steatosis Independent of the Arginine Content.

Nonalcoholic fatty liver disease, a major cause of abnormal liver function, is often associated with obesity. Arginine (ARG) plays a role in modulating body weight/fat, but limited data exist as to the role of ARG in soy protein's ability to protect from liver steatosis. We investigated the role of native ARG in the soy protein isolate (SPI) in reducing liver steatosis in male obese Zucker rats. Rats (N=48; 6 weeks old) were randomly assigned to one of three diets for 8 or 16 weeks: the casein (CAS) diet as control (0.6% ARG), CAS diet supplemented to contain 1.3% ARG, or an SPI diet containing isoflavones (1.3% ARG). SPI and ARG rats gained significantly more weight (P<.05) than CAS rats after 16 weeks only. The SPI rats had lower liver steatosis scores after 8 and 16 weeks (P<.05 and P<.001, respectively) compared to CAS and ARG rats. SPI rats had lower serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels (P<.05) compared to CAS after 16 weeks, and AST was lower (P<.05) compared to ARG rats. After 16 weeks, the SPI rats had lower (P<.05) serum ALT and AST levels than at 8 weeks. Our results suggest that a longer period of SPI feeding results in lower liver steatosis and serum ALT and AST levels, while the ARG diet had no effect on steatosis or ALT and AST levels. We found that the SPI diet reduced (P<.001) serum tumor necrosis factor-α (TNF-α) compared to CAS and ARG diets after 8 and 16 weeks. The SPI diet significantly reduced (P<.001) interleukin-6 (IL-6) when compared to the CAS diet at 8 weeks, but there was no significant difference at 16 weeks. Based on the findings of our study, the protective effect of SPI in reducing liver steatosis is not modulated by its native arginine content.