Spirooxadiazoline-oxindoles derived from imatinib show antimyeloproliferative potential in K562 cells.

Imatinib mesylate was the first representative BCR-ABL1 tyrosine kinase inhibitor (TKI) class for the treatment of chronic myeloid leukemia. Despite the revolution promoted by TKIs in the treatment of this pathology, a resistance mechanism occurs against all BCR-ABL1 inhibitors, necessitating a constant search for new therapeutic options. To develop new antimyeloproliferative substances, we applied a medicinal chemistry tool known as molecular hybridization to design 25 new substances. These compounds were synthesized and biologically evaluated against K562 cells, which express BCR-ABL1, a constitutively active tyrosine kinase enzyme, as well as in WSS-1 cells (healthy cells). The new compounds are conjugated hybrids that contain phenylamino-pyrimidine-pyridine (PAPP) and an isatin backbone, which are the main pharmacophoric fragments of imatinib and sunitinib, respectively. A spiro-oxindole nucleus was used as a linker because it occurs in many compounds with antimyeloproliferative activity. Compounds 2a, 2b, 3c, 4c, and 4e showed promise, as they inhibited cell viability by between 45% and 61% at a concentration of 10 µM. The CC50 of the most active substances was determined to be within 0.8-9.8 µM.

A Framework for Real-Time Gestural Recognition and Augmented Reality for Industrial Applications.

In recent decades, technological advancements have transformed the industry, highlighting the efficiency of automation and safety. The integration of augmented reality (AR) and gesture recognition has emerged as an innovative approach to create interactive environments for industrial equipment. Gesture recognition enhances AR applications by allowing intuitive interactions. This study presents a web-based architecture for the integration of AR and gesture recognition, designed to interact with industrial equipment. Emphasizing hardware-agnostic compatibility, the proposed structure offers an intuitive interaction with equipment control systems through natural gestures. Experimental validation, conducted using Google Glass, demonstrated the practical viability and potential of this approach in industrial operations. The development focused on optimizing the system's software and implementing techniques such as normalization, clamping, conversion, and filtering to achieve accurate and reliable gesture recognition under different usage conditions. The proposed approach promotes safer and more efficient industrial operations, contributing to research in AR and gesture recognition. Future work will include improving the gesture recognition accuracy, exploring alternative gestures, and expanding the platform integration to improve the user experience.

Antifungal activity of sustainable histone deacetylase inhibitors against planktonic cells and biofilms of Candida spp. and Cryptococcusneoformans.

The limited therapeutic options for fungal infections and the increased incidence of fungal strains resistant to antifungal drugs, especially Candida spp., require the development of new antifungal drugs and strategies. Histone deacetylase inhibitors (HDACi), like vorinostat, have been studied in cancer treatment and have antifungal effects, acting alone or synergistically with classical antifungals. Here we investigated the antifungal activity of two novel sustainable HDACi (LDT compounds) based on vorinostat structure. Molecular docking simulation studies reveal that LDT compounds can bind to Class-I HDACs of Candida albicans, C. tropicalis, and Cryptococcus neoformans, which showed similar binding mode to vorinostat. LDT compounds showed moderate activity when tested alone against fungi but act synergistically with antifungal azoles against Candida spp. They reduced biofilm formation by more than 50% in C. albicans (4 µg/mL), with the main action in fungal filamentation. Cytotoxicity of the LDT compounds against RAW264.7 cells was evaluated and LDT536 demonstrated cytotoxicity only at the concentration of 200 µmol/L, while LDT537 showed IC50 values of 29.12 µmol/L. Our data indicated that these sustainable and inexpensive HDACi have potential antifungal and antibiofilm activities, with better results than vorinostat, although further studies are necessary to better understand the mechanism against fungal cells.

Fungal infections are neglected diseases that affect more than a billion people worldwide. Some histone deacetylase inhibitors can act against fungal cells. Our data reveal that HDACi LDT536 and LDT537 have potential antibiofilm and antifungal activities.

Potential of yeasts as biocontrol agents against Fusarium graminearum in vitro and on corn.

AIMS: The antifungal effect of the yeast species Kluyveromyces marxianus, Meyerozyma caribbica, and Wickerhamomyces anomalus was evaluated against two Fusarium graminearum strains (FRS 26 and FSP 27) in vitro and on corn seeds. METHODS AND RESULTS: The antifungal effect of the yeasts against F. graminearum was evaluated using scanning electron microscopy and extracellular chitinase and glucanase production to further elucidate the biocontrol mode of action. In addition, the germination percentage and vigor test were investigated after applying yeast on corn seeds. All the yeast strains inhibited fungal growth in vitro (57.4%-100.0%) and on corn seeds (18.9%-87.2%). In co-culture with antagonistic yeasts, F. graminearum showed collapsed hyphae and turgidity loss, which could be related to the ability of yeasts to produce chitinases and glucanases. The three yeasts did not affect the seed corn germination, and W. anomalus and M. caribbica increased corn seed growth parameters (germination percentage, shoot and root length, and shoot dry weight). CONCLUSION: Meyerozyma caribbica and W. anomalus showed satisfactory F. graminearum growth inhibition rates and did not affect seed growth parameters. Further studies are required to evaluate the application of these yeasts to the crop in the field.

Trajectory of blood pressure levels and weight gain in pregnant women: A group-based approach.

BACKGROUND AND OBJECTIVES: Morbidities associated with increased blood pressure levels during pregnancy represent one of the main causes of maternal mortality. The objective was to identify patterns of systolic blood pressure (SPB) trajectory in pregnant women undergoing prenatal care at the Unified Health System, and associations with weight gain trajectory, demographic, obstetric, anthropometric data, and health related behaviors. METHODS AND RESULTS: Cohort study with pregnant women using the public health services in Brazil. Data were collected through questionnaires and medical records. Trajectory patterns of SBP and weight gain were identified by a group-based trajectory model. For trajectory analysis, 460 women had SBP information available, totaling 2839 measurements, with an average of 6.2 measurements during pregnancy. Three SBP trajectory patterns were identified and classified as "Group 1" (48.0%), with a mean of 103 mmHg (95% CI 102.5-103.7 mmHg), "Group 2" (42.7%) with a mean of 114 mmHg (95% CI 113.7-114.9 mmHg), and "Group 3" (9.1%) with the highest mean SBP value of 130 mmHg (95% CI 128.8-131.5 mmHg). It was observed that regardless of the weight gain trajectory group, women classified in the group with the highest SBP had the highest SBP levels. The probability of being classified in Group 3 was higher among women with higher education, who started pregnancy presenting obesity, and who were using antihypertensive drugs. CONCLUSION: The probability of belonging to groups with a greater trajectory of SBP during pregnancy was associated with obesity, education, and hypertension under treatment.

Depletion of TAX1BP1 Amplifies Innate Immune Responses during Respiratory Syncytial Virus Infection.

Respiratory syncytial virus (RSV) is the main cause of acute respiratory infections in young children and also has a major impact on the elderly and immunocompromised people. In the absence of a vaccine or efficient treatment, a better understanding of RSV interactions with the host antiviral response during infection is needed. Previous studies revealed that cytoplasmic inclusion bodies (IBs), where viral replication and transcription occur, could play a major role in the control of innate immunity during infection by recruiting cellular proteins involved in the host antiviral response. We recently showed that the morphogenesis of IBs relies on a liquid-liquid-phase separation mechanism depending on the interaction between viral nucleoprotein (N) and phosphoprotein (P). These scaffold proteins are expected to play a central role in the recruitment of cellular proteins to IBs. Here, we performed a yeast two-hybrid screen using RSV N protein as bait and identified the cellular protein TAX1BP1 as a potential partner of this viral protein. This interaction was validated by pulldown and immunoprecipitation assays. We showed that TAX1BP1 suppression has only a limited impact on RSV infection in cell cultures. However, RSV replication is decreased in TAX1BP1-deficient (TAX1BP1 knockout [TAX1BP1KO]) mice, whereas the production of inflammatory and antiviral cytokines is enhanced. In vitro infection of wild-type or TAX1BP1KO alveolar macrophages confirmed that the innate immune response to RSV infection is enhanced in the absence of TAX1BP1. Altogether, our results suggest that RSV could hijack TAX1BP1 to restrain the host immune response during infection. IMPORTANCE Respiratory syncytial virus (RSV), which is the leading cause of lower respiratory tract illness in infants, remains a medical problem in the absence of a vaccine or efficient treatment. This virus is also recognized as a main pathogen in the elderly and immunocompromised people, and the occurrence of coinfections (with other respiratory viruses and bacteria) amplifies the risks of developing respiratory distress. In this context, a better understanding of the pathogenesis associated with viral respiratory infections, which depends on both viral replication and the host immune response, is needed. The present study reveals that the cellular protein TAX1BP1, which interacts with the RSV nucleoprotein N, participates in the control of the innate immune response during RSV infection, suggesting that the N-TAX1BP1 interaction represents a new target for the development of antivirals.

New Imatinib Derivatives with Antiproliferative Activity against A549 and K562 Cancer Cells.

Tyrosine kinase enzymes are among the primary molecular targets for the treatment of some human neoplasms, such as those in lung cancer and chronic myeloid leukemia. Mutations in the enzyme domain can cause resistance and new inhibitors capable of circumventing these mutations are highly desired. The objective of this work was to synthesize and evaluate the antiproliferative ability of ten new analogs that contain isatins and the phenylamino-pyrimidine pyridine (PAPP) skeleton, the main pharmacophore group of imatinib. The 1,2,3-triazole core was used as a spacer in the derivatives through a click chemistry reaction and gave good yields. All the analogs were tested against A549 and K562 cells, lung cancer and chronic myeloid leukemia (CML) cell lines, respectively. In A549 cells, the 3,3-difluorinated compound (3a), the 5-chloro-3,3-difluorinated compound (3c) and the 5-bromo-3,3-difluorinated compound (3d) showed IC50 values of 7.2, 6.4, and 7.3 µM, respectively, and were all more potent than imatinib (IC50 of 65.4 µM). In K562 cells, the 3,3-difluoro-5-methylated compound (3b) decreased cell viability to 57.5% and, at 10 µM, showed an IC50 value of 35.8 µM (imatinib, IC50 = 0.08 µM). The results suggest that 3a, 3c, and 3d can be used as prototypes for the development of more potent and selective derivatives against lung cancer.

Effectiveness of fluoride-containing toothpastes associated with different technologies to remineralize enamel after pH cycling: an in vitro study.

BACKGROUND: To evaluate the efficacy of fluoride-containing toothpastes with different technologies to remineralize artificial caries lesions in enamel. METHODS: Bovine enamel blocks were divided into three thirds: intact (untreated), demineralized (artificial caries lesion), and treated (caries lesion, pH cycling with dentifrices). Enamel blocks were randomly distributed into five groups (n = 12): Fluoride-free toothpaste, Colgate Oral Care (NC); Arginine-containing toothpaste, Colgate Total Daily Repair (PC); Silicate-based fluoride toothpaste: REFIX technology, regenerador + sensitive (RDC), NR-5 technology, Regenerate Enamel Science (RES), and NOVAMIN technology, Sensodyne Repair and Protect (SRP). The specimens were submitted to a pH cycling model for 6 days. The efficacy of the toothpastes was estimated by calculating the surface microhardness recovery (%SMHR) and the fluorescence recovery (ΔFRE) with quantitative light-induced fluorescence. The cross-sectional micromorphology of the enamel surface was also assessed using scanning electron microscopy. Elemental analyses (weight%) were determined with an energy-dispersive X-ray spectrometer (EDS). The results were compared to that of the control (NC). Data were statistically analyzed (5%). RESULTS: %SMHR could be ranked as follows: RDC = PC = RES = SRP > NC. Significantly higher %SMHR and ΔFRE means were observed after enamel treatment with RDC (22.7 and 46.9, respectively). PC (%SMHR = 18.8) was as efficacious as RDC to recover the surface microhardness with a significantly lower mean of ΔFRE (19.5). Only RDC was able to promote the formation of a mineralized layer on the surface of enamel enriched with silicon on the surface. CONCLUSIONS: The silicate-based fluoride toothpaste containing REFIX technology demonstrated greater efficacy in the remineralizing artificial caries than the other products.

Nanoencapsulated fluoride as a remineralization option for dental erosion: an in vitro study.

OBJECTIVE: To compare the in vitro performance of different dentifrices indicated for dental erosion and a new dentifrice with controlled fluoride release system (NanoF) in terms of surface microhardness remineralization in enamel erosion lesions. MATERIALS AND METHODS: 72 human enamel specimens were divided into 6 groups (n = 12): PC (100% NaF - positive control); NC (Placebo - negative control); 50%nF (50% NanoF + 50% free NaF), 100%nF (100% NanoF); PN (Sensodyne® ProNamel™) and AG (Colgate® Sensitive Pro-Relief™). A surface microhardness analysis was performed before (SH0) and after (SH1) the erosion lesion formation. The blocks were submitted to a 5-day de-remineralization cycling model, consisting of 90 s immersion on 0.1% citric acid (4x/day) and 1 min treatment with dentifrice slurries along with 1 mL/block of human saliva (2x/day). Lastly, the final surface microhardness analysis (SH2) was measured and the percentage of surface microhardness remineralization (%SMHR) was calculated. Data were analysed with 2-way ANOVA and Tukey's test (p < .05). RESULTS: Statistically significant differences were observed for SH2 and %SMHR between NC and AG with the other groups (p < .05). The best %SMHR from the experimental groups was found in 100%nF and PN. CONCLUSION: Dentifrices with NanoF exhibited a surface microhardness remineralization similar to sodium fluoride (PC). Therefore, NanoF dentifrice can be an alternative to prevent and treat patients with dental erosion.

ELOVL5 Participates in Embryonic Lipid Determination of Cellular Membranes and Cytoplasmic Droplets.

Embryonic lipids are crucial for the formation of cellular membranes and dynamically participate in metabolic pathways. Cells can synthesize simple fatty acids, and the elongation of fatty acids facilitates the formation of complex lipids. The aim of this work was to investigate the involvement of the elongation of very long chain fatty acid enzyme 5 (ELOVL5) in embryonic development and lipid determination. Bovine embryos were produced in vitro using a standard protocol and randomly divided to receive one of three treatments at Day 4: morpholino (Mo) gene expression knockdown assay for ELOVL5 (ELOVL5-Mo), Mo antisense oligonucleotides for the thalassemic ß-globulin human mRNA (technical control Mo), and placebo (biological control). The phenotypes of embryonic development, cell number, ELOVL5 protein abundance, lipid droplet deposits, and lipid fingerprint were investigated. No detrimental effects (p > 0.05) were observed on embryo development in terms of cleavage (59.4 ± 3.5%, 63.6 ± 4.1%, and 65.4 ± 2.2%), blastocyst production (31.3 ± 4.2%, 28.1 ± 4.9%, and 36.1 ± 2.1%), and blastocyst cell number (99.6 ± 7.7, 100.2 ± 6.2, 86.8 ± 5.6), respectively, for biological control, technical control Mo, and ELOVL5-Mo. ELOVL5 protein abundance and cytoplasmic lipid droplet deposition were increased (p < 0.05) in ELOVL5-Mo-derived blastocysts compared with the controls. However, seven lipid species, including phosphatidylcholines, phosphatidylethanolamines, and triacylglycerol, were downregulated in the ELOVL5-Mo-derived blastocysts compared with the biological control. Therefore, ELOVL5 is involved in the determination of embryonic lipid content and composition. Transient translational blockage of ELOVL5 reduced the expression of specific lipid species and promoted increased cytoplasmic lipid droplet deposition, but with no apparent deleterious effect on embryonic development and blastocyst cell number.

Cashew Nut Shell Liquid (CNSL) as a Source of Drugs for Alzheimer's Disease.

Alzheimer's disease (AD) is a complex neurodegenerative disorder with a multifaceted pathogenesis. This fact has long halted the development of effective anti-AD drugs. Recently, a therapeutic strategy based on the exploitation of Brazilian biodiversity was set with the aim of discovering new disease-modifying and safe drugs for AD. In this review, we will illustrate our efforts in developing new molecules derived from Brazilian cashew nut shell liquid (CNSL), a natural oil and a byproduct of cashew nut food processing, with a high content of phenolic lipids. The rational modification of their structures has emerged as a successful medicinal chemistry approach to the development of novel anti-AD lead candidates. The biological profile of the newly developed CNSL derivatives towards validated AD targets will be discussed together with the role of these molecular targets in the context of AD pathogenesis.

Dental arch perimeter changes as a result from premature loss of primary anterior teeth due to trauma: A case series in infant and pre-school children.

BACKGROUND: The premature loss of primary anterior teeth in deciduous arches is a controversial topic in the literature, especially due to the lack of robust scientific evidence about the consequences in the arch perimeter space and magnitudes of the effects involved. AIM: Evaluate the association between premature loss of primary anterior teeth and dental arch perimeter changes, according to clinical variables as deciduous arch type, erupted primary canines, midline involvement and deleterious oral habits, on infants and pre-school children. DESIGN: Patients with avulsion or referral to extraction due to traumatic dental injuries (TDI) were evaluated. After the tooth loss, two trained operators measured the tooth/teeth space, both with a digital caliper and a dry tip compass. The clinical documentation included photographs and radiographs. Follow-up visits occurred from the baseline and every two months over a 12-month period. Chi-square test was used to evaluate the association between arch perimeter changes and clinical variables (α = 0.05). A descriptive statistic was performed to explore the magnitude of space changes, with 95% confidence intervals. RESULTS: Eighteen infants/children (mean, 2.78 ± 1.39 years) were included. Nine patients presented space loss (50.0%) (mean, -1.32 mm), six patients gained space (33.3%) (mean, +1.55 mm), and three patients presented space maintenance (16.7%). Clinical variables did not influence dental arch perimeter changes. CONCLUSIONS: Premature loss of primary anterior teeth, as well as deleterious oral habits, deciduous arch type, midline involvement and erupted primary canines, were not associated with dental arch perimeter changes.

(Phenylamino)pyrimidine-1,2,3-triazole derivatives as analogs of imatinib: searching for novel compounds against chronic myeloid leukemia.

The enzyme tyrosine kinase BCR-Abl-1 is the main molecular target in the treatment of chronic myeloid leukemia and can be competitively inhibited by tyrosine kinase inhibitors such as imatinib. New potential competitive inhibitors were synthesized using the (phenylamino)pyrimidine-pyridine (PAPP) group as a pharmacophoric fragment, and these compounds were biologically evaluated. The synthesis of twelve new compounds was performed in three steps and assisted by microwave irradiation in a 1,3-dipolar cycloaddition to obtain 1,2,3-triazole derivatives substituted on carbon C-4 of the triazole nucleus. All compounds were evaluated for their inhibitory activities against a chronic myeloid leukemia cell line (K562) that expresses the enzyme tyrosine kinase BCR-Abl-1 and against healthy cells (WSS-1) to observe their selectivity. Three compounds showed promising results, with IC50 values between 1.0 and 7.3 µM, and were subjected to molecular docking studies. The results suggest that such compounds can interact at the same binding site as imatinib, probably sharing a competitive inhibition mechanism. One compound showed the greatest interaction affinity for BCR-Abl-1 in the docking studies.

NEK10 interactome and depletion reveal new roles in mitochondria.

BACKGROUND: Members of the family of NEK protein kinases (NIMA-related kinases) were described to have crucial roles in regulating different aspects of the cell cycle. NEK10 was reported to take part in the maintenance of the G2/M checkpoint after exposure to ultraviolet light. NEK1, NEK5, NEK2 and NEK4 proteins on the other hand have been linked to mitochondrial functions. METHODS: HEK293T cells were transfected with FLAG empty vector or FLAG-NEK10 and treated or not with Zeocin. For proteomic analysis, proteins co-precipitated with the FLAG constructs were digested by trypsin, and then analyzed via LC-MS/MS. Proteomic data retrieved were next submitted to Integrated Interactome System analysis and differentially expressed proteins were attributed to Gene Ontology biological processes and assembled in protein networks by Cytoscape. For functional, cellular and molecular analyses two stable Nek10 silenced HeLa cell clones were established. RESULTS: Here, we discovered the following possible new NEK10 protein interactors, related to mitochondrial functions: SIRT3, ATAD3A, ATAD3B, and OAT. After zeocin treatment, the spectrum of mitochondrial interactors increased by the proteins: FKBP4, TXN, PFDN2, ATAD3B, MRPL12, ATP5J, DUT, YWHAE, CS, SIRT3, HSPA9, PDHB, GLUD1, DDX3X, and APEX1. We confirmed the interaction of NEK10 and GLUD1 by proximity ligation assay and confocal microscopy. Furthermore, we demonstrated that NEK10-depleted cells showed more fragmented mitochondria compared to the control cells. The knock down of NEK10 resulted further in changes in mitochondrial reactive oxygen species (ROS) levels, decreased citrate synthase activity, and culminated in inhibition of mitochondrial respiration, affecting particularly ATP-linked oxygen consumption rate and spare capacity. NEK10 depletion also decreased the ratio of mtDNA amplification, possibly due to DNA damage. However, the total mtDNA content increased, suggesting that NEK10 may be involved in the control of mtDNA content. CONCLUSIONS: Taken together these data place NEK10 as a novel regulatory player in mitochondrial homeostasis and energy metabolism.

Checking NEKs: Overcoming a Bottleneck in Human Diseases.

In previous years, several kinases, such as phosphoinositide 3-kinase (PI3K), mammalian target of rapamycin (mTOR), and extracellular-signal-regulated kinase (ERK), have been linked to important human diseases, although some kinase families remain neglected in terms of research, hiding their relevance to therapeutic approaches. Here, a review regarding the NEK family is presented, shedding light on important information related to NEKs and human diseases. NEKs are a large group of homologous kinases with related functions and structures that participate in several cellular processes such as the cell cycle, cell division, cilia formation, and the DNA damage response. The review of the literature points to the pivotal participation of NEKs in important human diseases, like different types of cancer, diabetes, ciliopathies and central nervous system related and inflammatory-related diseases. The different known regulatory molecular mechanisms specific to each NEK are also presented, relating to their involvement in different diseases. In addition, important information about NEKs remains to be elucidated and is highlighted in this review, showing the need for other studies and research regarding this kinase family. Therefore, the NEK family represents an important group of kinases with potential applications in the therapy of human diseases.

Erosive potential of soy-based beverages on dental enamel.

BACKGROUND: This study aimed to investigate the erosive potential of soy-based beverages in comparison to fruit juices of the same flavor. METHODS: Human enamel blocks were randomly divided into 9 groups (n = 8), according to the beverage category (soy or non-soy juices). The initial pH, TA and ß at the original pH value were measured in triplicate. The composition of calcium, phosphate and total protein was analyzed using the specific colorimetric method. The fluoride analysis was performed using a selective electrode. The degree of saturation (DS) and the critical pH (CpH) of each beverage with respect to hydroxyapatite (HAp) and fluorapatite (FAp) were calculated using the computational software. Enamel samples were immersed into 67.5 mL of each drink for 120 minutes. Enamel surface loss (ESL) and differences in surface roughness (ΔRaE-S) were analyzed by a 3D non-contact profilometer. RESULTS: Non-soy beverages exhibited the lowest pH values (2.93 to 3.40). The highest values of calcium concentration were founded in soy-based formulations. Juices with soy in their composition tend to have high DS when compared with non-soy based beverages (p = .0571). Soy beverages produced less ESL than non-soy beverages (p < .05). ΔRaE-S was not significantly different between the categories. The ESL and ΔRaE-S were positively correlated with initial pH and buffering capacity in soy-based beverages. On the other hand, in non-soy beverages, the ESL was negatively correlated with the TA to 7.0 and the fluoride composition whereas the ΔRaE-S was negatively correlated with the TA to 5.5. CONCLUSIONS: The erosive potential of soy beverages was lower than non-soy based beverages.

In situ effect of a CPP-ACP chewing gum on enamel erosion associated or not with abrasion.

OBJECTIVES: The purpose of this study is to analyze the in situ effect of a casein phosphopeptide-stabilized amorphous calcium phosphate (CPP-ACP) chewing gum on human enamel erosion lesion associated or not with abrasion. MATERIAL AND METHODS: A three-way crossover study of 7 days was conducted involving 10 volunteers subjected to the same protocol: (G1) CPP-ACP sugar-free chewing gum, (G2) regular sugar-free chewing gum without CPP-ACP, and (G3) saliva-no chewing gum. An abrasion test was included in each phase. A 3D non-contact profilometry measurement of lesion depth and surface roughness was obtained of sound and eroded surfaces. A salivary calcium concentration was determined for all volunteers. ANOVA followed by Tukey's test were used with a p < 0.05. RESULTS: The enamel depth and the enamel surface roughness of the CPP-ACP gum group were significantly lower than the others (ANOVA, p < 0.05). No significant differences were observed between the treatments when associated with abrasion (p > 0.05). A positive and significant correlation was seen between the lesion depth and enamel surface roughness for GI (r = 0.87, p = 0.00) and GIII (r = 0.79, p = 0.00) groups. The estimated total calcium presented in the saliva after the chewed CPP-ACP gum showed no statistical significance between the mean absorbance values at the different time collections (p > 0.05). CONCLUSIONS: It is demonstrated that the incorporation of the CPP-ACP into a sugar-free gum significantly increased the remineralization/protection of eroded enamel surface. CLINICAL RELEVANCE: The CPP-ACP added to gum may be a suitable alternative vehicle, to deliver calcium ions to saliva and therefore protecting enamel.

Interactome analysis of the human respiratory syncytial virus RNA polymerase complex identifies protein chaperones as important cofactors that promote L-protein stability and RNA synthesis.

UNLABELLED: The human respiratory syncytial virus (HRSV) core viral RNA polymerase comprises the large polymerase protein (L) and its cofactor, the phosphoprotein (P), which associate with the viral ribonucleoprotein complex to replicate the genome and, together with the M2-1 protein, transcribe viral mRNAs. While cellular proteins have long been proposed to be involved in the synthesis of HRSV RNA by associating with the polymerase complex, their characterization has been hindered by the difficulty of purifying the viral polymerase from mammalian cell culture. In this study, enhanced green fluorescent protein (EGFP)-tagged L- and P-protein expression was coupled with high-affinity anti-GFP antibody-based immunoprecipitation and quantitative proteomics to identify cellular proteins that interacted with either the L- or the P-proteins when expressed as part of a biologically active viral RNP. Several core groups of cellular proteins were identified that interacted with each viral protein including, in both cases, protein chaperones. Ablation of chaperone activity by using small-molecule inhibitors confirmed previously reported studies which suggested that this class of proteins acted as positive viral factors. Inhibition of HSP90 chaperone function in the current study showed that HSP90 is critical for L-protein function and stability, whether in the presence or absence of the P-protein. Inhibition studies suggested that HSP70 also disrupts virus biology and might help the polymerase remodel the nucleocapsid to allow RNA synthesis to occur efficiently. This indicated a proviral role for protein chaperones in HRSV replication and demonstrates that the function of cellular proteins can be targeted as potential therapeutics to disrupt virus replication. IMPORTANCE: Human respiratory syncytial virus (HRSV) represents a major health care and economic burden, being the main cause of severe respiratory infections in infants worldwide. No vaccine or effective therapy is available. This study focused on identifying those cellular proteins that potentially interact specifically with the viral proteins that are central to virus replication and transcription, with a view to providing potential targets for the development of a specific, transient therapeutic which disrupts virus biology but prevents the emergence of resistance, while maintaining cell viability. In particular, protein chaperones (heat shock proteins 70 and 90), which aid protein folding and function, were identified. The mechanism by which these chaperones contribute to virus biology was tested, and this study demonstrates to the field that cellular protein chaperones may be required for maintaining the correct folding and therefore functionality of specific proteins within the virus replication complex.

Exposure assessment of dogs to mycotoxins through consumption of dry feed.

BACKGROUND: The Brazilian exportation of pet food has shown high growth rates in the last two years and determination of the exposure degree is one of the most important parameters for the risk assessment of chemical compounds. In this study the exposure degree of dogs to mycotoxins was estimated and acceptable daily intake (ADI) and safe pet dietary level (SPDL) were calculated. Thus the natural occurrence of fumonisins, zearalenone and aflatoxins was evaluated in 100 dry dog feed samples provided by pet owners in Paraná State, Brazil. RESULTS: Despite the high frequency of fumonisins (68%), zearalenone (95%) and aflatoxins (68%) in feed samples, the mean levels detected were low. ADI for fumonisins and zearalenone was 20.0 and 1.00 µg kg(-1) body weight (BW) day(-1) respectively and SPDL for fumonisins was 2000 µg kg(-1) feed. The probable daily intake values (1.83 µg fumonisins, 0.93 µg zearalenone and 0.02 µg aflatoxins kg(-1) BW day(-1) ) were low. CONCLUSION: The exposure degree of dogs could be assumed to be very low. However, the co-occurrence of these three or other mycotoxins, and possible synergic or additive effects, should be taken into account when determining the maximum allowed levels or risk assessment. © 2016 Society of Chemical Industry.

Development and validation of a food frequency questionnaire for consumption of polyphenol-rich foods in pregnant women.

Previous studies have shown that maternal consumption of polyphenol-rich foods after the third trimester of pregnancy may interfere with the anatomical and functional activity of the fetal heart as, to our knowledge, there are no validated instruments to quantify total polyphenols in pregnant women. The aim of this study was evaluate the reproducibility and validity of a food frequency questionnaire (FFQ), with 52 items, to assess the intake of polyphenol-rich foods in pregnant women in Brazil. This cross-sectional study included 120 pregnant women who participated in nutritional interviews in two moments. The intake of polyphenols estimated by the developed FFQ was compared with the average of two 24-h recalls (24HR), with the average intake measured by a 3-day food diary (D3days) and with the urinary excretion of total polyphenols. The triangular method was applied to calculate Pearson's correlation coefficients, intraclass correlation and Bland-Altman plots for the FFQ, using an independent biochemical marker, in addition to classification by quarters of consumption. The questionnaires were log transformed, adjusted for body mass index and gestational age. The adjustment for energy was applied only of 24HR and D3days. Analysis of the reproducibility between the FFQ showed a very high correlation (r = 0.72; P < 0.05). A low but significant association was observed between the FFQ and urinary excretion (0.23; P = 0.01). The association between the dietary survey methods was moderate to very high (r = 0.36 to r = 0.72; P < 0.001). In conclusion, this questionnaire showed reproducibility and validity for the quantification of consumption of total polyphenols in pregnant women.