Establishment of methods to analyze the structural and functional integrity of the quail (Coturnix coturnix japonica) sperm plasma membrane.

1. The objectives of this study were to establish the use of the fluorophores Hoechst 33342 and propidium iodide for the evaluation of sperm plasma membrane integrity and to identify an adequate hypoosmotic solution for the evaluation of sperm membrane functionality in quails.2. Sperm samples were collected from the vas deferens of nine quails. After initial evaluation, the samples were subjected to a flash-frozen assay. Three treatments with the following proportions of fresh sperm and sperm subjected to flash freezing were prepared as follows: 100:0 (T100), 50:50 (T50), and 0:100 (T0). The hypoosmotic swelling test used distilled water (0 mOsm/l) and fructose solutions (50, 100, and 200 mOsm/l).3. Immediately after recovery, the samples showed 75.6 ± 5.0% motility with vigour of 3.7 ± 0.3 and 96.1 ± 0.5% of the sperm appeared normal. The membrane integrity test showed 62.2 ± 5.2% intact sperm at T100, 29.0 ± 4.1% at T50 and 0.1 ± 0.1% at T0. Moreover, a greater number of reactive sperm (74.7 ± 6.7%) were observed when incubated in distilled water (0 mOsm/l) in comparison to other solutions (P < 0.05).4. The association of fluorescent probes composed of Hoechst 33342 and propidium iodide provided an efficient assessment of the integrity of the plasmatic membrane of quail spermatozoa. However, the study identified that the hypoosmotic swelling test has little predictive value regarding sperm membrane functionality in this species.

Effect of glycoelectrolytic supplement on post-weaning piglet performance and intestinal integrity.

This study evaluated the effects of glycoelectrolytic supplements on the performance, blood parameters, and intestinal morphology of piglets during the post-weaning period. In the study, a total of 240 piglets weaned aged 17 22.60 + 1.10 days were used. The control group (n = 120) received only water, and the treatment group (n = 120) received an oral glycoelectrolytic supplement diluted in water (0.75%) during the first three days after weaning. Feed intake, daily weight gain, final weight, feed conversion ratio, and post-weaning mortality were evaluated. On the third day after weaning, the blood glucose levels of all piglets were analyzed. Blood was collected from 12 piglets from each treatment group on day 3 after weaning for blood count analysis, and intestinal fragments were collected for anatomopathological and morphometric evaluation. Better feed conversion ratio (1.29) and higher consumption of liquids (0.639 L/day) were observed in the piglet group supplemented with glycoelectrolytes on day 3 after weaning (P < 0.05). The supplemented group presented a higher glycemic index (80.78 mg/dL), average corpuscular volume (67.35 fL), and average corpuscular hemoglobin (20.46 pg) than the control group (P < 0.05). The evaluation of intestinal integrity and the probability of diarrhea occurrence were similar between the groups (P > 0.05). Oral glycoelectrolytic supplementation can be an option for piglets immediately after weaning as it improves feed conversion and consumption of liquids, in addition to increasing blood glucose without the occurrence of diarrhea, thus reducing dehydration and energy deficit.

IL-1ß-pre-conditioned mesenchymal stem/stromal cells' secretome modulates the inflammatory response and aggrecan deposition in intervertebral disc.

Mesenchymal stem/stromal cells (MSCs) have been increasingly used in clinical trials for low-back pain (LBP) and intervertebral disc (IVD) degeneration with promising results. Their action mechanisms are not fully understood, but they reduce IVD pro-inflammatory markers in a pro-inflammatory/degenerative IVD microenvironment. In this study the therapeutic potential of the MSC secretome, as an alternative cell-free approach for treating degenerated IVDs, was examined. Human bone marrow-derived MSC secretome (MSCsec) was collected after 48 h of preconditioning in IL-1ß (10 ng/mL) and low oxygen (6 % O2), mimicking the degenerative IVD. IL-1ß-pre-conditioning of MSCs increased secretion of pro-inflammatory markers hIL-6, hIL-8, hMCP-1, etc. The therapeutic effect of MSCsec was tested in a pro-inflammatory/degenerative IVD ex vivo model. MSCsec down-regulated IVD gene expression of pro-inflammatory cytokines (bIL-6, bIL-8) and matrix degrading enzyme bMMP1, while bMMP3 and bTIMP2 were up-regulated, at 48 h. After 14 d, MSCsec-treated IVDs revealed increased aggrecan deposition, although no differences in other ECM components were observed. Protein analysis of the MSCsec-treated IVD supernatant revealed a significant increase of CXCL1, MCP-1, MIP-3α, IL-6, IL-8 and GRO α/ß/γ (related to TNF, NOD-like receptor and neutrophil chemotaxis signalling), and a decrease of IFN-γ, IL-10, IL-4, IL-5 and TNF-α (associated with T-cell receptor signalling). MSCsec-treated IVD supernatants did not promote angiogenesis and neurogenesis in vitro. Overall, MSCsec can be a safe therapeutic approach, presenting a strong immunomodulatory role in degenerated IVD while potentiating aggrecan deposition, which can open new perspectives on the use of MSCsec as a cell-based/ cell-free therapeutic approach to LBP.

Brachio-cervical inflammatory myopathy associated with systemic sclerosis. Case series and review of literature.

This study was aimed at describing a case series of brachio-cervical inflammatory myopathy (BCIM) associated with systemic sclerosis (SSc), due to its rarity and limited coverage in published data. Another aim was to provide a literature review. We reported four cases of BCIM-SSc from our tertiary center. In addition, we researched the literature and found six articles featuring 17 patients who fit this phenotype. We pooled all cases and reported their features. Most patients were female and had limited SSc, and the median time of BCIM presentation was three years after SSc diagnosis. Asymmetric muscle involvement, scapular winging, dropped head, axial weakness, camptocormia, dysphagia, and dermatomyositis stigmas were common features. All patients had esophageal involvement. Most had positive antinuclear antibody results, a scleroderma pattern in their capillaroscopy images, elevated serum creatine phosphokinase, myopathic electrophysiology, and muscle involvement in magnetic resonance imaging. Muscle histopathological findings varied widely, but in general all showed the presence of lymphoid infiltrates, muscle atrophy, increased MHC-I expression, MAC deposits, vasculopathy, and muscle fiber necrosis. The response to immunosuppressive therapy was highly irregular. BCIM-SSc is a rare disorder that shares many similar phenotypes among the described cases, but has a highly heterogeneous response to treatment. At present, more data on the physiopathology, clinical features, and treatment is still needed.

Composition of semen and foreskin mucosa aerobic microbiota and its impact on sperm parameters of captive collared peccaries (Pecari tajacu).

AIM: To evaluate the bacterial composition of collared peccary semen and foreskin mucosa, and to verify the sensitivity of isolates to antimicrobials used in semen conservation and to Aloe vera gel, which is an alternative external cryoprotectant. METHODS AND RESULTS: Nine foreskin mucosa and ejaculate samples from adult animals were used. Sperm characteristics and bacterial load were evaluated in fresh semen. The preputial mucosa and semen bacterial isolates were identified and tested against five concentrations of each antimicrobial (streptomycin-penicillin and gentamicin) and A. vera gel. Corynebacterium sp. and Staphylococcus sp. were isolated in greater numbers than others in both semen (64·10 and 20·51%, respectively) and the foreskin mucosa (60·60 and 24·25%, respectively), and ranged from 0·4 to 21 × 105 colony-forming units (CFU) per ml. The average load of Corynebacterium sp. was negatively correlated (P < 0·05) with the sperm membrane integrity (r = -0·73055) and curvilinear velocity (r = -0·69048). Streptomycin-penicillin and gentamicin inhibited most micro-organisms, and A. vera showed lower antimicrobial activity. CONCLUSION: Several Gram-positive bacteria are present in semen and foreskin mucosa of collared peccary, and the benefits of using primarily penicillin-streptomycin and gentamicin antimicrobials in the bacterial control of diluted semen of these animals are strongly indicated. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides insight into the reproductive microbiota of captive male-collared peccary. This work provides a theoretical basis to assist reproductive biotechnologies for ex situ conservation of the species.

Risk Factors Associated with Probable Sleep Bruxism of Children and Teenagers with Cerebral Palsy.

AIM: To assess the association between probable sleep bruxism (PSB) and other occlusal characteristics in children and teenagers with Cerebral Palsy (CP). STUDY DESIGN: A cross-sectional study was carried out with 148 participants (74 with CP and 74 without special needs) aged between 2 and 14 years old. Participants underwent an oral clinical examination to evaluate the occlusal characteristics. Parents/caregivers filled out a questionnaire with information related to the typical sounds of PSB, sociodemographic factors and the presence of harmful oral habits in the study participants. Data analysis was carried out, using Chi-square or Fisher's exact test and Odds Ratio (p<0.05). RESULTS: PSB and malocclusion (68.9% and 95.9%, respectively) were more prevalent in participants with CP than in participants without CP. The association between PSB and the presence of a wear facet was statistically significant (p < 0.001) in participants with CP. In this group, 64.8% of participants with PSB presented tooth wear. CONCLUSION: Presence of dental wear facets was significantly associated with PSB in individuals with cerebral palsy.

Effects of pre- or post-exercise whey protein supplementation on body fat and metabolic and inflammatory profile in pre-conditioned older women: A randomized, double-blind, placebo-controlled trial.

BACKGROUND AND AIM: Protein supplementation and resistance training (RT) are interventions that may counteract decline in muscle mass and increase in fat mass, thus reducing the risk of developing chronic diseases during the aging process. The objective of this study was to investigate the effect of whey protein (WP) pre- or post-RT on metabolic and inflammatory profile in pre-conditioned older women. METHODS AND RESULTS: Seventy older women participated in this investigation and were randomly assigned to one of three groups: WP pre-RT and placebo post-RT (WP-PLA, n = 24), placebo pre-RT and WP post-RT (PLA-WP, n = 23) and placebo pre and post-RT (PLA-PLA, n = 23). Each group ingested 35 g of PLA or WP pre- and post-RT. RT was carried out over 12 weeks (three times/week; 3 x 8-12 repetition maximum). Body composition, blood pressure, blood samples and dietary intake were assessed pre- and post-intervention. After the intervention, WP groups showed greater improvements in appendicular lean soft tissue (ALST: WP-PLA, 3.1%; PLA-WP, 3.9%; PLA-PLA, 1.8%) and total cholesterol/high density lipoprotein cholesterol ratio (TC/HDL-C: WP-PLA, -12.11%; PLA-WP, -13.2%; PLA-PLA, -0.7) when compared with PLA-PLA. WP post-RT also showed improvements (P < 0.05) in ALST/appendicular fat mass ratio (PLA-WP, 5.8%; PLA-PLA, 1.3%), total body fat (PLA-WP, -3.8%; PLA-PLA: -0.1) and trunk fat mass (PLA-WP, -3.1%; PLA-PLA, -0.3%) when compared with PLA-PLA. CONCLUSION: WP pre- or post- RT promotes improvements in ALST and TC/HDL-C ratio in pre-conditioned older women. WP administered after RT was more effective in improving metabolic health Z-score and in reducing body fat compared to placebo group.

Biological and molecular characterization of two Anticarsia gemmatalis multiple nucleopolyhedrovirus clones exhibiting contrasting virulence.

Baculovirus natural populations are known to be genetically heterogeneous and such genotypic diversity could have implications in the performance of biocontrol agents. The Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV) has been widely used to control the velvetbean caterpillar, Anticarsia gemmatalis, in Brazil. In the present work, morphological and molecular analyses as well as the biological activity of AgMNPV genotypes derived from a Brazilian field isolate (AgMNPV-79) were carried out. The existence of genotypic variants in the population was confirmed by DNA restriction analysis. Although difference in virulence was observed among the variants, the most (Ag79-01) and the least (AgL-16) virulent clones do not show any morphological and cytopathological changes when compared to the most studied isolate (AgMNPV-2D). The complete genome analysis of the two viral clones showed the presence of single open reading frames (ORFs) of the pe-38 and he65 genes, which contrasts with the two split ORFs present in the genome of the AgMNPV-2D isolate. The viral clone AgL-16 has many variations in the ie-2 and pe-38 genes, which are transcription regulatory genes responsible for the regulation of viral early gene expression during insect cell infection. Furthermore, other genes showed alterations like the odv-e56, which have an essential role in the maturation and envelopment of the ODVs, and bro-a and bro-b genes which were fused to form a single ORF. For the Ag79-01, although the total number of single nucleotide variants (SNVs) was more prominent in the pe-38 gene, its genome showed very few modifications in comparison to the AgMNPV-2D genome.

Short communication: In vitro and in vivo probiotic potential of Lactobacillus plantarum B7 and Lactobacillus rhamnosus D1 isolated from Minas artisanal cheese.

Some Lactobacillus strains may contribute to the health of the host when administered in adequate concentrations, demonstrating their probiotic potential. In contrast, Listeria monocytogenes is a foodborne pathogen that can cause enteropathy, meningoencephalitis, abortion, and septicemia. The aim of this survey was to evaluate the in vitro and in vivo probiotic potential of Lactobacillus plantarum B7 and Lactobacillus rhamnosus D1, isolated from Minas artisanal cheese of the Serra da Canastra (Minas Gerais, Brazil), against Lis. monocytogenes. We submitted B7 and D1 to in vitro testing (antibiogram, tolerance to bile salts and artificial gastric fluid, and spot-on-lawn) and in vivo testing (relative weight gain in mice). Both Lactobacillus strains demonstrated in vitro inhibitory activity against Lis. monocytogenes, as well as sensitivity to antimicrobials and resistance to gastric acids and bile salts. In the in vivo assays, mice treated with D1 gained more weight than mice in the other groups. These results indicate that D1 could have higher probiotic potential than B7 because improvements in feed conversion may help animals fight infection.

3D printing in the design of pharmaceutical dosage forms.

Three-dimensional (3D) printing technologies are manufacturing approaches with widespread use in industry (e.g. automotive, automobile, pharmaceutical industries). With regard to its use in pharmaceutical industry, 3D printing is demonstrating to be of added value attributed to the possibility of printing tailored pharmaceutical products, namely personalized medical devices, such as implants and other dosage forms. However, with the approval of the first 3D-printed drug-product in 2015, a new perspective has arisen, i.e. the use of this technology to produce solid oral dosage forms exhibiting complex drug release profiles and allowing for individual dosing. Technological hurdles and regulatory issues still have to be overcome before this technology can truly find its place in the healthcare sector, where it can certainly contribute to a personalized and patient-centered healthcare. This manuscript offers a comprehensive analysis of the most extensively used methods of 3D printing in the pharmaceutical field, with examples of solid oral dosage forms and other medical devices currently under development or already marketed.

A functional glycoproteomics approach identifies CD13 as a novel E-selectin ligand in breast cancer.

BACKGROUND: The glycan moieties sialyl-Lewis-X and/or -A (sLeX/A) are the primary ligands for E-selectin, regulating subsequent tumor cell extravasation into distant organs. However, the nature of the glycoprotein scaffolds displaying these glycans in breast cancer remains unclear and constitutes the focus of the present investigation. METHODS: We isolated glycoproteins that bind E-selectin from the CF1_T breast cancer cell line, derived from a patient with ductal carcinoma. Proteins were identified using bottom-up proteomics approach by nanoLC-orbitrap LTQ-MS/MS. Data were curated using bioinformatics tools to highlight clinically relevant glycoproteins, which were validated by flow cytometry, Western blot, immunohistochemistry and in-situ proximity ligation assays in clinical samples. RESULTS: We observed that the CF1_T cell line expressed sLeX, but not sLeA and the E-selectin reactivity was mainly on N-glycans. MS and bioinformatics analysis of the targeted glycoproteins, when narrowed down to the most clinically relevant species in breast cancer, identified CD44 glycoprotein (HCELL) and CD13 as key E-selectin ligands. Additionally, the co-expression of sLeX-CD44 and sLeX-CD13 was confirmed in clinical breast cancer tissue samples. CONCLUSIONS: Both CD44 and CD13 glycoforms display sLeX in breast cancer and bind E-selectin, suggesting a key role in metastasis development. Such observations provide a novel molecular rationale for developing targeted therapeutics. GENERAL SIGNIFICANCE: While HCELL expression in breast cancer has been previously reported, this is the first study indicating that CD13 functions as an E-selectin ligand in breast cancer. This observation supports previous associations of CD13 with metastasis and draws attention to this glycoprotein as an anti-cancer target.

Meta-analysis of the energy and protein requirements of hair sheep raised in the tropical region of Brazil.

The objective of this study was to estimate, through mathematical models, energy and protein requirements for maintenance and gain of hair sheep raised in the tropical region of Brazil. To determine the equation parameters, a meta-analysis of seven independent experiments of nutrient requirements was performed, comprising a total of 243 experimental units (animals), which were conducted under tropical conditions, using hair sheep in growing and finishing phases and endowed of the following quantitative data for each animal: body weight (BW), empty body weight (EBW), average daily gain (ADG), empty body gain (EBG), heat production (HP), metabolizable energy intake (MEI), retained energy (RE), metabolizable protein intake (MPI) and body protein content. The regression equations generated were as follows: for Net Energy for maintenance, (NEm ): LogHP(MJEBW-0.75day-1)=-0.6090(±0.07470)+0.5149(±0.07216)×MEI(MJEBW-0.75day-1); for Net Energy for gain, (NEg ): LogRE(MJEBW-0.75day-1)=0.03084(±0.05334)+0.8455(±0.04355)×LogEBG(kg/day); for Metabolizable Protein for maintenance,(MPm ): MPI(g/day)  = 24.8470 (±7.3646) + 560.28 (±99.6582) × EBG(kg/day) ; for Net Protein for gain, (NPg ): NPg(kg/day)=0.1941×EBW(kg)-0.1058. The NEm requirement was 0.246 MJ EBW-0.75  day-1 . The metabolizable energy for maintenance requirement was 0.391 MJ EBW-0.75  day-1 . Considering an ADG of 100 g, the NEg requirement ranged from 0.496 to 1.701 MJ/day for animals with BW ranging from 10 to 40 kg respectively. The efficiencies of use of the metabolizable energy for maintenance and gain were 0.63 and 0.36 respectively. The MPm requirement was 3.097 g EBW-0.75  day-1 . Considering an ADG of 100 g, the NPg requirement ranged from 12.4 to 10.5 g/day for animals with BW ranging from 10 to 40 kg respectively. The total metabolizable energy and protein requirements were lower than those reported by the NRC and AFRC systems. Thus, our results support the hypothesis that nutrient requirements of hair sheep raised in tropical regions differ from wool sheep raised in temperate regions. Therefore, the use of the equations designed in this study is recommended.

Effect of resistance training on phase angle in older women: A randomized controlled trial.

Phase angle (PhA) is an angular-derived bioelectrical impedance parameter based on resistance and reactance that has been widely used in different populations as an objective indicator of cellular health. The purpose of this study was to analyze the effect of resistance training (RT) on PhA in older women. Forty-one older women (67.2 ± 4.5 years, 64.0 ± 12.3 kg, 154.7 ± 5.2 cm, and 26.6 ± 4.8 kg/m2 ) were randomly assigned to one of two groups: a training group (TG, n = 19) that underwent progressive RT for 12 weeks (eight exercises, three sets of 10-15 repetitions maximum, three times per week) or a control group (CG, n = 22) that did not perform any type of exercise during the intervention period. Resistance, reactance, PhA, total body water (TBW), intracellular (ICW) and extracellular (ECW) water were assessed by spectral bioelectrical impedance. There was a significant group by time interaction (P < 0.05) for PhA, TBW, and ICW in which only the TG increased their scores after the intervention period (PhA = +6.5%, TBW = +2.8%, and ICW = +5.1%). The results suggest that progressive RT promotes an increase in PhA in older women. Therefore, the PhA seems to be a good parameter to assess changes in cellular health during RT intervention.

Solid lipid nanoparticles affect microbial colonization and enzymatic activity throughout the decomposition of alder leaves in freshwater microcosms.

Solid lipid nanoparticles (SLNs) are used as carriers for drug delivery, and are high biocompatible and designed to endure in the host organism. Despite its current industrial production is low, many of these substances are available on the market, and much more are in the production pipeline. As a result, many of them will end in aquatic systems raising the question whether they can pose a risk to aquatic biota and the associated ecological processes. Microbial decomposers of plant litter, play a key role in forested streams being responsible for the energy flow between terrestrial and aquatic environments. Here, we investigated the effects of SLNs on alder leaf litter decomposition by aquatic microbes. Alder leaves were immersed in a stream of Northeast Portugal to allow microbial colonization before being exposed in microcosms of two types of SLNs at two concentrations for 42 days. Results showed that rates of leaf decomposition decreased with exposure to SLNs. Bacterial biomass was not inhibited by SLNs, and cultivable fungi densities remained constant (SLN-A) or increased (SLN-C) compared with control microcosms. The type and concentration of SLNs influenced differently the leaf colonization by fungi as well as fungal sporulation rate. These effects were accompanied by changes in the community extraenzymatic profile: the activities of alkaline phosphatase, acidic phosphatase, Naphthol-AS-BI-phosphohydrolase (P cycle) and lipases increased in the SLNs microcosms. This study provided the first evidence of the adverse effects of the release of SLNs to streams on leaf litter decomposition. Those effects seem to depend on the composition and concentration of SLNs, as well on the microbial target group, or enzyme. Thus, prior to massive industrial production of these nanomaterials, some measures should be taken to avoid environmental impact affecting the microbial communities responsible for detritus decomposition.

Hypertrophy-type Resistance Training Improves Phase Angle in Young Adult Men and Women.

The main purpose of the present study was to investigate the effect of a hypertrophy-type resistance training protocol on phase angle, an indicator of cellular integrity, in young adult men and women. 28 men (22.2±4.3 years, 67.8±9.0 kg and 174.2±6.8 cm) and 31 women (23.2±4.1 years, 58.7±12.1 kg and 162.7±6.4 cm) underwent a progressive RT for 16 weeks (2 phases, 8 weeks each), 3 times per week, consisting of 10 to 12 whole body exercises with 3 sets of 8-12 repetitions maximum. Phase angle, resistance, reactance and total body water (intra and extracellular water compartments) were assessed by bioimpedance spectroscopy (Xitron 4200 Bioimpedance Spectrum Analyzer). Total body water, intracellular water and phase angle increased significantly (P<0.05) in men (7.8, 8.3, and 4.3%, respectively) and women (7.6, 11.7, and 5.8% respectively), with no significant difference between sexes (P>0.05). Bioimpedance resistance decreased (P<0.05) similarly in both sex (men=-4.8%, women=-3.8%). The results suggest that regardless of sex, progressive RT induces an increase in phase angle and a rise in cellular hydration.

Mendelian inheritance, genetic linkage, and genotypic disequilibrium for nine microsatellite loci in Cariniana estrellensis (Raddi) Kuntze (Lecythidaceae).

Cariniana estrellensis is one of the largest trees found in Brazilian tropical forests. The species is typical of advanced stages of succession, characteristic of climax forests, and essential in genetic conservation and environmental restoration plans. In this study, we assessed Mendelian inheritance, genetic linkage, and genotypic disequilibrium in nine microsatellite loci for a C. estrellensis population. We sampled and genotyped 285 adult trees and collected seeds from 20 trees in a fragmented forest landscape in Brazil. Based on maternal genotypes and their seeds, we found no deviation from the expected 1:1 Mendelian segregation and no genetic linkage between pairwise loci. However, for adults, genotypic disequilibrium was detected for four pairs of loci, suggesting that this result was not caused by genetic linkage. Based on these results, we analyzed microsatellite loci that are suitable for use in population genetic studies assessing genetic diversity, mating system, and gene flow in C. estrellensis populations.

Supervised toothbrushing programs in primary schools and early childhood settings: A scoping review.

OBJECTIVE: In this article we report the findings of a scoping review that aimed to identify and summarise the range of programs and guidelines available for toothbrushing programs in schools and early childhood settings. Dental caries is one of the most common preventable diseases affecting children worldwide. Untreated caries can impact on child health and wellbeing, development, socialisation and school attendance. Supervised toothbrushing programs in schools and other early childhood settings can be effective in improving the oral health of young children. There is limited understanding of the salient issues to consider when developing such programs or how they are best implemented in real world settings. METHODS: A scoping review methodology was utilised to provide a summary of the guidelines and programs available. Key search terms were developed, mapped and utilised to identify guidelines and programs across 6 databases and key search engines. RESULTS: We located 26 programs and guidelines that met the inclusion and exclusion criteria for the review. These were collated and summarised across key countries and critical aspects of program development and implementation were identified. Toothbrush type and storage, toothpaste strength and method of dispensing, toothbrush storage, staff training and parental consent are key considerations that varied widely. CONCLUSIONS AND RECOMMENDATIONS: Guidelines for supervised toothbrushing programs vary within and across countries due to differences in water fluoridation and availability of low fluoride toothpastes. The results of this review provide critical information to be considered when establishing and implementing toothbrushing programs in these settings.

d-α-tocopherol nanoemulsions: Size properties, rheological behavior, surface tension, osmolarity and cytotoxicity.

The aim of this study was the assessment of the physicochemical stability of d-α-tocopherol formulated in medium chain triglyceride nanoemulsions, stabilized with Tween®80 and Lipoid®S75 as surfactant and co-surfactant, respectively. d-α-tocopherol was selected as active ingredient because of its well-recognized interesting anti-oxidant properties (such as radical scavenger) for food and pharmaceutical industries. A series of nanoemulsions of mean droplet size below 90 nm (polydispersity index < 0.15) have been produced by high-pressure homogenization, and their surface electrical charge (zeta potential), pH, surface tension, osmolarity, and rheological behavior, were characterized as a function of the d-α-tocopherol loading. In vitro studies in Caco-2 cell lines confirmed the safety profile of the developed nanoemulsions with percentage of cell viability above 90% for all formulations.

Ibuprofen nanocrystals developed by 22 factorial design experiment: A new approach for poorly water-soluble drugs.

The reduction of the particle size of drugs of pharmaceutical interest down to the nano-sized range has dramatically changed their physicochemical properties. The greatest disadvantage of nanocrystals is their inherent instability, due to the risk of crystal growth. Thus, the selection of an appropriate stabilizer is crucial to obtain long-term physicochemically stable nanocrystals. High pressure homogenization has enormous advantages, including the possibility of scaling up, lack of organic solvents and the production of small particles diameter with low polydispersity index. The sequential use of high shear homogenization followed by high pressure homogenization, can modulate nanoparticles' size for different administration routes. The present study focuses on the optimization of the production process of two formulations composed of different surfactants produced by High Shear Homogenization followed by hot High Pressure Homogenization. To build up the surface response charts, a 22 full factorial design experiment, based on 2 independent variables, was used to develop optimized formulations. The effects of the production process on the mean particle size and polydispersity index were evaluated. The best ibuprofen nanocrystal formulations were obtained using 0.20% Tween 80 and 1.20% PVP K30 (F1) and 0.20% Tween 80 and 1.20% Span 80 (F2). The estimation of the long-term stability of the aqueous suspensions of ibuprofen nanocrystals was studied using the LUMISizer. The calculated instability index suggests that F1 was more stable when stored at 4 °C and 22 °C, whereas F2 was shown to be more stable when freshly prepared.

Lactic acid microbiota identification in water, raw milk, endogenous starter culture, and fresh Minas artisanal cheese from the Campo das Vertentes region of Brazil during the dry and rainy seasons.

Minas artisanal cheese, produced in the Campo das Vertentes region of Brazil, is made from raw milk and endogenous starter cultures. Although this cheese is of great historical and socioeconomic importance, little information is available about its microbiological and physical-chemical qualities, or about its beneficial microbiota. This work was aimed at evaluating the qualities of the cheese and the components used for its production, comparing samples collected during the dry and rainy seasons. We also conducted molecular identification and isolated 50 samples of lactic acid bacteria from cheese (n=21), water (n=3), raw milk (n=9), and endogenous starter culture (n=17). The microbiological quality of the cheese, water, raw milk, and endogenous starter culture was lower during the rainy period, given the higher counts of coagulase-positive Staphylococcus and total and thermotolerant coliforms. Enterococcus faecalis was the lactic acid bacteria isolated most frequently (42.86%) in cheese samples, followed by Lactococcus lactis (28.57%) and Lactobacillus plantarum (14.29%). Lactobacillus brevis (5.88%), Enterococcus pseudoavium (5.88%), Enterococcus durans (5.88%), and Aerococcus viridans (5.88%) were isolated from endogenous starter cultures and are described for the first time in the literature. The lactic acid bacteria identified in the analyzed cheeses may inhibit undesirable microbiota and contribute to the safety and flavor of the cheese, but this needs to be evaluated in future research.