The absorption of bovine milk small extracellular vesicles largely depends on galectin 3 and galactose ligands in human intestinal cells and C57BL/6J mice.

Small extracellular vesicles in milk (sMEVs) have attracted attention in drug delivery and as bioactive food compounds. Previous studies implicate galactose residues on the sMEV surface in sMEV transport across intestinal and endothelial barriers in humans, but details of glycoprotein-dependent transport are unknown. We used a combination of cell biology and genetics protocols to identify glycoproteins on the sMEV surface that facilitate sMEV absorption. We identified 256 proteins on the bovine sMEVs surface by using LC-MS/MS, and bioinformatics analysis suggested that 42, 13, and 13 surface proteins were N-, O-, and 13 C-glycosylated, respectively. Lectin blots confirmed the presence of mannose, galactose, N-acetyl galactose, fucose, and neuraminate. When surface proteins were removed by various treatment with various proteases, sMEV uptake decreased by up to 58% and 67% in FHs-74 Int and Caco-2 cells, respectively, compared with controls (P < 0.05). When glycans were removed by treatment with various glycosidases, sMEV uptake decreased by up to 54% and 74% in FHs-74 Int and Caco-2 cells, respectively (P < 0.05). When galactose and N-acetyl galactosamine residues were blocked with agglutinins, sMEV uptake decreased by more than 50% in FHs-74 Int cells (P < 0.05). When bovine sMEVs were administered to Galectin-3 knockout mice by oral gavage, hepatic sMEV accumulation decreased by 56% compared with wild-type mice (P < 0.05), consistent with a role of ß-galactoside glycan structures in the absorption of sMEVs. We conclude that sMEVs are decorated with glycoproteins, and Galectin-3 and its galactose ligands are particularly important for sMEV absorption.NEW & NOTEWORTHY This is the first paper to assess the role of unique glycans and their Galectin-3 receptor in the transport and distribution of small extracellular vesicles ("exosomes") from milk in mammals. The research assessed milk exosome transport and distribution by using multiple approaches and platforms including cell cultures, various exosome labels, knockout and mutant mice, enzymatic removal of surface proteins and glycans, and lectin blocking of glycans.

Comparative polar and lipid plasma metabolomics differentiate KSHV infection and disease states.

BACKGROUND: Kaposi sarcoma (KS) is a neoplastic disease etiologically associated with infection by the Kaposi sarcoma-associated herpesvirus (KSHV). KS manifests primarily as cutaneous lesions in individuals due to either age (classical KS), HIV infection (epidemic KS), or tissue rejection preventatives in transplantation (iatrogenic KS) but can also occur in individuals, predominantly in sub-Saharan Africa (SSA), lacking any obvious immune suppression (endemic KS). The high endemicity of KSHV and human immunodeficiency virus-1 (HIV) co-infection in Africa results in KS being one of the top 5 cancers there. As with most viral cancers, infection with KSHV alone is insufficient to induce tumorigenesis. Indeed, KSHV infection of primary human endothelial cell cultures, even at high levels, is rarely associated with long-term culture, transformation, or growth deregulation, yet infection in vivo is sustained for life. Investigations of immune mediators that distinguish KSHV infection, KSHV/HIV co-infection, and symptomatic KS disease have yet to reveal consistent correlates of protection against or progression to KS. In addition to viral infection, it is plausible that pathogenesis also requires an immunological and metabolic environment permissive to the abnormal endothelial cell growth evident in KS tumors. In this study, we explored whether plasma metabolomes could differentiate asymptomatic KSHV-infected individuals with or without HIV co-infection and symptomatic KS from each other. METHODS: To investigate how metabolic changes may correlate with co-infections and tumorigenesis, plasma samples derived from KSHV seropositive sub-Saharan African subjects in three groups, (A) asymptomatic (lacking neoplastic disease) with KSHV infection only, (B) asymptomatic co-infected with KSHV and HIV, and (C) symptomatic with clinically diagnosed KS, were subjected to analysis of lipid and polar metabolite profiles RESULTS: Polar and nonpolar plasma metabolic differentials were evident in both comparisons. Integration of the metabolic findings with our previously reported KS transcriptomics data suggests dysregulation of amino acid/urea cycle and purine metabolic pathways, in concert with viral infection in KS disease progression. CONCLUSIONS: This study is, to our knowledge, the first to report human plasma metabolic differentials between in vivo KSHV infection and co-infection with HIV, as well as differentials between co-infection and epidemic KS.

First Isolation and Genome Sequence Analysis of West Nile Virus in Mosquitoes in Brazil.

West Nile virus is a flavivirus transmitted by mosquitoes, mainly of the genus Culex. In Brazil, serological studies have already indicated the circulation of the virus since 2003, with the first human case detected in 2014. The objective of the present paper is to report the first isolation of WNV in a Culex (Melanoconion) mosquito. Arthropods were collected by protected human attraction and CDC light bait, and taxonomically identified and analyzed by viral isolation, complement fixation and genomic sequencing tests. WNV was isolated from samples of Culex (Melanoconion) mosquitoes, and the sequencing analysis demonstrated that the isolated strain belonged to lineage 1a. The finding of the present study presents the first evidence of the isolation and genome sequencing of WNV in arthropods in Brazil.

Collection of Plasma Samples in Areas with Limited Healthcare Access.

Regular monitoring of various biomarkers and molecular panels in plasma can significantly help to prevent disease onset and improve its management and final outcomes. Many groups can benefit from monitoring programs focusing on the prevention of cardiovascular diseases, evaluation of environmental exposure impacts, or the prevention/management of cancer. Improvement in therapeutic options in part due to targeted therapeutic agents and monoclonal antibody therapies has led to a significant sized population that can be described as "cancer survivors." These patients, although in remission from their original disease, are at significant risk for the recurring disease and must be monitored for adverse events. Monitoring is, however, not an easy task; requiring a high level of complexity in lab facilities and blood/plasma sampling, collection, and storage must occur under tightly controlled conditions. These demanding circumstances are especially difficult to attain in rural areas and in historically marginalized populations. The Telimmune Plasma Separation Card (TPS card or TPSC) has been developed to enable diagnostic plasma sampling, collection, and stabilization in locations that may be remote to laboratory or clinic. The TPSC requires a drop of blood applied to a top of a separation system consisting of a separation membrane and collection disk. In 3 min, the TPSC device separates plasma from erythrocytes and deposits a defined volume of plasma into a collection disc which is air-dried for 15 min to deliver a stabilized, volumetric plasma sample, which may be stored or shipped at ambient temperatures with minimal biological risk. Extraction of proteins and metabolites is then achieved in well-equipped laboratories using protocols discussed in this chapter.

The effects of mercury exposure on Amazonian fishes: An investigation of potential biomarkers.

Exposure to mercury can interfere with the expression of proteins and enzymes, compromise important pathways, such as apoptosis and glucose metabolism, and even induce the expression of metallothioneins. In this study, analytical techniques were used to determine the concentration of total mercury (THg) in muscle and liver tissue, protein pellets, and spots [using graphite furnace atomic absorption spectrometry (GFAAS)], and molecular techniques were used to identify metalloproteins present in mercury-associated protein spots. Thirty individuals from three different fish species, Cichla sp. (n = 10), Brachyplatystoma filamentosum (n = 10), and Semaprochilodus sp. (n = 10) from the Brazilian Amazon were used. Oxidative stress indicators [such as glutathione peroxidase (GSH-Px), catalase (CAT), superoxide dismutase (SOD), a marker of lipid peroxidation (LPO)] and the possible expression of metallothioneins in muscle and liver tissues were investigated. The two piscivorous species, Cichla sp. and B. filamentosum, presented the highest concentrations of mercury in their hepatic tissue, 1219 ± 15.00 and 1044 ± 13.6 µg kg-1, respectively, and in their muscle tissue, 101 ± 1.30 µg kg-1 and 87.4 ± 0.900 µg kg-1, respectively. The non-carnivorous species Semaprochilodus sp. had comparatively low concentrations of mercury in both its hepatic (852 ± 11.1 µg kg-1) and muscle (71.4 ± 0.930 µg kg-1) tissues. The presence of mercury was identified in 24 protein spots using GFAAS; concentrations ranged from 11.5 to 787 µg kg-1, and mass spectrometry identified 21 metal-binding proteins. The activities of GSH-Px, CAT, and SOD, related to oxidative stress, decreased proportionally as tissue Hg concentrations increased, while the levels of LPO markers increased, indicating the presence of stress. Our study results demonstrate possible mercury interference in oxidative stress markers (GSH-Px, CAT, SOD, and LPO), in addition to the identification of 21 metal-binding proteins as possible biomarkers of mercury exposure in fish.

Metalloproteomic approach to liver tissue of rats exposed to mercury.

The aims of this study were to identify mercury-associated protein spots in the liver tissue of rats exposed to low concentrations of mercury and to elucidate the physiological and functional aspects of the proteins identified in the protein spots. Therefore, proteomic analysis of the liver tissue of Wistar rats exposed to mercury chloride (4.60 µg kg-1 in Hg2+) was performed for thirty days (Hg-30 group) and sixty days (Hg-60 group). The proteomic profile of the liver tissue of the rats was obtained by two-dimensional electrophoresis (2D-PAGE), and the determinations of total mercury in the liver tissue, pellets and protein spots were performed by graphite furnace atomic absorption spectrometry (GFAAS). ImageMaster 2D Platinum 7.0 software was used to identify the differentially expressed mercury-associated protein spots, which were then characterized by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The determinations by GFAAS indicated a total mercury bioaccumulation of 2812% in the Hg-30 group and 3298% in the Hg-60 group and 10 mercury-associated protein spots with a concentration range of 51 ± 1.0 to 412 ± 6.00 mg kg-1 in the 2D PAGE gels from the liver tissue of the Hg-60 group. The LC-MS/MS analyses allowed the identification of 11 metal binding proteins in mercury-associated protein spots that presented fold change with upregulation >1.5, downregulation < -1.7 or that were expressed only in the Hg-60 group. Using the FASTA sequences of the proteins identified in the mercury-associated protein spots, bioinformatics analyses were performed to elucidate the physiological and functional aspects of the metal binding proteins, allowing us to infer that enzymes such as GSTM2 presented greater mercury concentrations and downregulation < -3; Acaa2 and Bhmt, which showed expression only in the Hg-60 group, among others, may act as potential mercury exposure biomarkers.

Metalloproteomic Investigation of Hg-Binding Proteins in Renal Tissue of Rats Exposed to Mercury Chloride.

Results obtained from rat studies indicate that, even at low concentrations, mercurial species cause harmful effects on the kidneys, by inducing the nephrotic oxidative stress response. In the present work, Hg-associated proteins were identified as possible mercury-exposure biomarkers in rat kidneys exposed to low mercury chloride concentrations for 30 days (Hg-30) and 60 days (Hg-60), using metalloproteomic strategies. The renal proteomic profile was fractioned by two-dimensional electrophoresis and the mercury determinations in kidney samples, protein pellets and protein spots were performed using graphite furnace atomic absorption spectrometry. The characterization of Hg-associated protein spots and the analysis of differentially expressed proteins were performed by liquid chromatography, coupled with tandem mass spectrometry. Eleven Hg-associated protein spots with a concentration range of 79 ± 1 to 750 ± 9 mg kg-1 in the Hg-60 group were identified. The characterization and expression analyses allowed the identification of 53 proteins that were expressed only in the Hg-60 group, 13 "upregulated" proteins (p > 0.95) and 47 "downregulated" proteins (p < 0.05). Actin isoforms and hemoglobin subunits were identified in protein spots of the Hg-60 group, with mercury concentrations in the range of 138 to 750 mg kg-1, which qualifies these proteins as potential mercury-exposure biomarkers.

Mercury metalloproteomic profile in muscle tissue of Arapaima gigas from the Brazilian Amazon.

Metalloproteomics is an innovative methodology for identifying of protein-associated mercury. Thus, we analyzed the muscle proteome of Arapaima gigas (pirarucu), collected in the Madeira River of the Brazilian Amazon, to identify protein-associated mercury, with the aim of identifying possible mercury biomarkers in fish muscle tissue. After obtaining the protein pellet, we conducted two-dimensional electrophoresis (2D PAGE) to fractionate the muscle proteome. Total mercury in muscle tissue and protein pellets and mapping of mercury content in protein spots of the 2D PAGE gels was determined using graphite furnace atomic absorption spectrometry (GFAAS). The protein-associated mercury identification was performed using liquid chromatography coupled with sequence mass spectrometry (LC‒MS/MS). Total mercury determinations by GFAAS indicated concentrations on the order of 153 ± 1.90 mg kg-1 and 142 ± 1.50 mg kg-1 (total precipitation of protein fraction) and 139 ± 1.45 mg kg-1 (fractional precipitation of protein fraction) in muscle tissue and protein pellets, respectively. Mercury concentrations in the range of 48 ± 0.90 to 165 ± 3.00 mg kg-1 were found in twelve protein spots. Among the 2D PAGE protein spots, eleven Hg-binding proteins were identified using LC‒MS/MS, which showed characteristics of mercury exposure biomarkers for important metabolic functions, such as five parvalbumin isoforms, triosephosphate isomerase, cofilin 2 (muscle), and fructose-bisphosphate aldolases.

Feedlot diets containing different starch levels and additives change the cecal proteome involved in cattle's energy metabolism and inflammatory response.

Diets for feedlot cattle must be a higher energy density, entailing high fermentable carbohydrate content. Feed additives are needed to reduce possible metabolic disorders. This study aimed to analyze the post-rumen effects of different levels of starch (25%, 35%, and 45%) and additives (monensin or a blend of essential oils and exogenous α-amylase) in diets for Nellore feedlot cattle. The cecum tissue proteome was analyzed via two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and then differentially expressed protein spots were identified with liquid chromatography-tandem mass spectrometry (LC-MS/MS). The use of blends of essential oils associated with α-amylase as a feed additive promoted the upregulation of enzymes such as triosephosphate isomerase, phosphoglycerate mutase, alpha-enolase, beta-enolase, fructose-bisphosphate aldolase, pyruvate kinase, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), L-lactate dehydrogenase B, L-lactate dehydrogenase A chain, L-lactate dehydrogenase, and ATP synthase subunit beta, which promote the degradation of carbohydrates in the glycolysis and gluconeogenesis pathways and oxidative phosphorylation, support pyruvate metabolism through the synthesis of lactate from pyruvate, and participate in the electron transport chain, producing ATP from ADP in the presence of a proton gradient across the membrane. The absence of proteins related to inflammation processes (leukocyte elastase inhibitors) in the cecum tissues of animals fed essential oils and amylase may be because feed enzymes can remain active in the intestine and aid in the digestion of nutrients that escape rumen fermentation; conversely, the effect of monensin is more evident in the rumen and less than 10% results in post-ruminal action, corroborating the hypothesis that ionophore antibiotics have a limited effect on the microbiota and intestinal fermentation of ruminants. However, the increase in starch in these diets promoted a downregulation of enzymes linked to carbohydrate degradation, probably caused by damage to the cecum epithelium due to increased responses linked to inflammatory injuries.

Ultrasonication of Milk Decreases the Content of Exosomes and MicroRNAs in an Exosome-Defined Rodent Diet.

BACKGROUND: Bovine milk exosomes (BMEs) harbor regulatory proteins, lipids, and microRNAs. Consumption of an exosome- and RNA-depleted (ERD) diet elicited phenotypes compared with controls fed an exosome- and RNA-sufficient (ERS) diet in mice. All other ingredients were identical in the diets. ERD and ERS diets were prepared by substituting ultrasonicated and nonultrasonicated milk, respectively, for casein in the AIN-93G formulation. OBJECTIVES: The objective of this study was to assess the effect of ultrasonication of milk on exosome content and bioavailability, and cargo content. METHODS: Bovine milk was ultrasonicated and exosomes were isolated by ultracentrifugation [ultrasonicated exosomes (USEs)]; controls were not ultrasonicated [nonultrasonicated exosomes (NSEs)]. Exosome count, size, and morphology were assessed using a nanoparticle tracker and electron microscopy. RNAs, lipids, and proteins were analyzed by RNA sequencing and MS. Intestinal transport, bioavailability, and distribution were measured by using fluorophore-labeled USEs and NSEs in Caco-2 cells, FHs 74 Int cells, and C57BL/6J mice (n = 3; age: 6-8 wk). RESULTS: The exosome count was 76% ± 22% lower in USEs than in NSEs (P < 0.05). Ultrasonication caused a degradation of ≤100% of microRNAs. USEs and NSEs contained 145 and 332 unique lipid signatures, respectively (P < 0.05). We detected a total of 525 and 484 proteins in USEs and NSEs, respectively. The uptake of USEs decreased by 46% ± 30% and 40% ± 27% compared with NSEs in Caco-2 and FHs 74 Int cells, respectively (P < 0.05). The hepatic accumulation of USEs was 48% ± 28% lower than the accumulation of NSEs in mice (P < 0.05). CONCLUSIONS: Ultrasonication of milk depletes bioavailable BMEs in studies of Caco-2 cells, FHs 74 Int cells, and C57BL/6J mice and causes a near-complete degradation of microRNA cargos.

Investigation of Protein Biomarkers and Oxidative Stress in Pinirampus pirinampu Exposed to Mercury Species from the Madeira River, Amazon-Brazil.

In recent decades, the scientific community has widely debated the contamination of fish in the Amazon region by mercury species. As the diet of riverside populations in the Amazon region is based mainly on fish, these populations are exposed to mercurial species that can cause serious and irreversible damage to their health. The risks of consuming fish exposed to mercurial species in the Amazon region have motivated toxicological investigations. However, the effect of mercurial species on protein and enzyme levels is still controversial. In this work, analytical and bioanalytical techniques Two-dimensional polyacrylamide gel electrophoresis [2D-PAGE] Graphite Furnace Atomic Absorption Spectrometry [GFAAS], and Mass Spectrometry in Sequence with Electrospray Ionization [ESI-MS/MS] were used to identify proteins associated with mercury (metal-binding protein) in muscle and liver tissues of the fish species Pinirampus pirinampu from the Madeira River, in the Brazilian Amazon. Enzymatic and lipid peroxidation analyses were also used to assess changes related to oxidative stress. Determinations of total mercury by GFAAS indicated higher concentrations in liver tissue (555 ± 19.0 µg kg-1) when compared to muscle tissue (60 ± 2.0 µg kg-1). The fractionation process of tissue proteomes by 2D-PAGE and subsequent mapping of mercury by GFAAS in the protein spots of the gels identified the presence of mercury in three spots of the liver tissue (concentrations in the range of 0.800 to 1.90 mg kg-1). The characterization of protein spots associated with mercury by ESI-MS/MS identified the enzymes triosephosphate isomerase A, adenylate kinase 2 mitochondrial, and glyceraldehyde-3-phosphate dehydrogenase as possible candidates for mercury exposure biomarkers. The muscle tissue did not show protein spots associated with mercury. Enzymatic activity decreased proportionally to the increase in mercury concentrations in the tissues.

Avaliação Qualitativa de Cardápios Hospitalares / Qualitative Evaluation of Hospital Menus

Modelo: Estudo caracteriza-se como transversal, qualitativo. Objetivo: Avaliar qualitativamente as preparações de cardápios oferecidos em hospitais no município de Uberaba - MG. Metodologia: A análise foi realizada pelo método "Avaliação Qualitativa de Preparações do Cardápio" dos almoços oferecidos em cinco Unidades de Alimentação e Nutrição hospitalares públicas ou particulares durante três semanas alternadas entre os meses de fevereiro a junho de 2018. Avaliou-se as ofertas de frutas, folhosos, repetições de cores, presença de alimentos sulfurados, doces, frituras, carnes gordurosas e frituras + doces na mesma refeição. Resultados: Os itens foram classificados como "ótimo" a baixa oferta de frituras (6,7%), carnes gordurosas (8,0%) e nenhuma oferta de doce + frituras no mesmo dia (0%), "bom" para oferta de folhosos (75%) e doces (22,7%), "regular" para repetição de cores (42.7%), "ruim" quanto aos alimentos sulfurados (56%) e "péssimos" quanto a oferta de frutas (9,3%). Conclusão: Os cardápios possuíam aspectos positivos como ausência de doce + fritura, baixa utilização de frituras e carnes gordurosas, boa oferta de folhosos e doces; os aspectos preocupantes relacionaram-se com a repetição de cores e negativos com a presença de alimentos sulfurados e ausência de frutas. (AU)

Study design: Cross-sectional, qualitative study. Objectives: To qualitatively evaluate the menu preparations offered at hospitals from the city of Uberaba - MG Methods: The analysis was conducted using the method "Qualitative evaluation of menu preparations" to evaluate lunch menus offered at 5 Food and Nutrition Units from public or private Hospitals for 3 weeks alternated between February and June 2018. The offer of fruits, leafy vegetables, sweets, fried food, fatty meats and fried food + sweets in the same meal were evaluated, as well as the color repetitions and the presence of sulphurated food. Results: Items rated as "Excellent" showed low offer of fried food (6.7%), fatty meats (8.0%) and no offer of sweet + fried food on the same day (0%); "Good" showed the offer of leafy vegetables (75%) and sweets (22.7%); "Regular" indicated color repetition (42.7%); "Bad" indicated sulphurated food (56%) and "Very bad" stood for fruit offer (9.3%). Conclusion: The menus had positive aspects such as the absence of sweet + fried food, low presence of fried food and fatty meats, good offer of leafy vegetables and sweets. The worrying aspects were associated with color repetition and the negative aspects were related to the presence of sulphurated food and absence of fruits. (AU)

Isolation of extracellular vesicles from byproducts of cheesemaking by tangential flow filtration yields heterogeneous fractions of nanoparticles.

Extracellular vesicles (EV) in milk, particularly exosomes, have attracted considerable attention as bioactive food compounds and for their use in drug delivery. The utility of small EV in milk (sMEV) as an animal feed additive and in drug delivery would be enhanced by cost-effective large-scale protocols for the enrichment of sMEV from byproducts in dairy plants. Here, we tested the hypothesis that sMEV may be enriched from byproducts of cheesemaking by tangential flow filtration (EV-FF) and that the sMEV have properties similar to sMEV prepared by ultracentrifugation (sMEV-UC). Three fractions of EV were purified from the whey fraction of cottage cheese making by using EV-FF that passed through a membrane with a 50-kDa cutoff (50 penetrate; 50P), and subfractions of 50P that were retained (100 retentate; 100R) or passed through (100 penetrate; 100P) a membrane with a 100-kDa cutoff; sMEV-UC controls were prepared by serial ultracentrifugation. The abundance of sMEV (<200 nm) was less than 0.3% in EV-FF compared with sMEV-UC (1012/mL of milk). Despite the low EV count, the protein content (mg/mL) of 100R (63 ± 0.02; ± standard deviation) was higher than that of 50P (0.75 ± 0.10), 100P (0.65 ± 0.40), and sMEV-UC (27 ± 0.02). There were 17, 14, 35, and 75 distinct proteins detected by nontargeted mass spectrometry analysis in 50P, 100R, 100P, and sMEV-UC, respectively. Exosome markers CD9, CD63, CD81, HSP-70, PDCD6IP, and TSG101 were detected in control sMEV-UC but not in EV-FF by using targeted mass spectrometry and immunoblot analyses. Negative exosome markers, APOB, ß-integrin, and histone H3 were below the limit of detection in EV-FF and control sMEV-UC analyzed by immunoblotting. The abundance of the major milk fat globule protein butyrophilin showed the following pattern: 100R ≫ 100P = 50P > sMEV-UC. More than 100 mature microRNA were detected in sMEV-UC by using sequencing analysis, compared with 36 to 60 microRNA in EV-FF. Only 100R and sMEV-UC yielded mRNA in quantities and qualities sufficient for sequencing analysis; an average of 276,000 and 838,000 reads were mapped to approximately 14,600 and 18,500 genes in 100R and sMEV-UC, respectively. In principal component analysis, microRNA, mRNA, and protein in EV-FF preparations clustered separately from control sMEV-UC. We conclude that under the conditions used here, flow filtration yields a heterogeneous population of milk EV.

An in silico, structural, and biological analysis of lactoferrin of different mammals.

Lactoferrin (LF) belongs to the family of transferrins having multifunctional roles associated with the immune system of animals. To follow the aims for this study was selected 20 sequences of LF from mammalian species to evaluate the chemical, biological, and structural properties. Bioinformatics approaches used programs such as MAFFT for sequence alignment; PartitionFinder and MrBayes for phylogenetic approaches; I-TASSER, PROCHECK, Molecular Operating Environment (MOE), SWISS Model server, Peptide DB and Expasy ProtParam to estimate the physicochemical properties, to model the protein and predicted secondary structures. A phylogenic analysis shows species with genetic similarities clustered by complexity and unique grouping between Capra hircus, Macaca mulatta, and Myotis lucifugus, since they presented more amino acids but not overall changes in the iron-binding sites or biological aspects. Structural deviations in these clusters obtained in LF from those species were found in residues 46 (position 406-450), that is part of alpha-helix, and 37 (position 295-331), that is part of the beta-sheets. Our predicted model can be used to investigate more about structural aspects of LF and be applied for medicinal research.

Microbiome and oral squamous cell carcinoma: a possible interplay on iron metabolism and its impact on tumor microenvironment.

There is increasing evidence showing positive association between changes in oral microbiome and the occurrence of oral squamous cell carcinoma (OSCC). Alcohol- and nicotine-related products can induce microbial changes but are still unknown if these changes are related to cancerous lesion sites. In an attempt to understand how these changes can influence the OSCC development and maintenance, the aim of this study was to investigate the oral microbiome linked with OSCC as well as to identify functional signatures and associate them with healthy or precancerous and cancerous sites. Our group used data of oral microbiomes available in public repositories. The analysis included data of oral microbiomes from electronic cigarette users, alcohol consumers, and precancerous and OSCC samples. An R-based pipeline was used for taxonomic and functional prediction analysis. The Streptococcus spp. genus was the main class identified in the healthy group. Haemophilus spp. predominated in precancerous lesions. OSCC samples revealed a higher relative abundance compared with the other groups, represented by an increased proportion of Fusobacterium spp., Prevotella spp., Haemophilus spp., and Campylobacter spp. Venn diagram analysis showed 52 genera exclusive of OSCC samples. Both precancerous and OSCC samples seemed to present a specific associated functional pattern. They were menaquinone-dependent protoporphyrinogen oxidase pattern enhanced in the former and both 3',5'-cyclic-nucleotide phosphodiesterase (purine metabolism) and iron(III) transport system ATP-binding protein enhanced in the latter. We conclude that although precancerous and OSCC samples present some differences on microbial profile, both microbiomes act as "iron chelators-like" potentially contributing to tumor growth.

Assessment of eating behavior and food neophobia in children and adolescents from uberaba-mg / Avaliação do comportamento alimentar e da neofobia alimentar em crianças e adolescentes do município de uberaba, mg

ABSTRACT Objective: To investigate and compare the eating behavior and food neophobia of children and adolescents from different age groups, body mass index per age, and sex. Methods: This was a cross-sectional study, with a convenience sample, involving 150 children and adolescents aged 3 to 13 years, of both sexes, treated at a pediatric outpatient clinic of a teaching hospital in the municipality of Uberaba-MG, Brazil. Subscales of the Child Eating Behavior Questionnaire (CEBQ) were used to evaluate eating behavior, and the Child Food Neophobia Scale (CFNS) was used to evaluate food neophobia. Results: Higher scores were found in the subscales "food responsiveness" (p=0.015), "enjoyment of food" (p=0.002), and "emotional overeating" (p=0.009) among older children and adolescents. Younger children had higher scores in the subscales "satiety responsiveness" (p=0.004) and "slowness in eating" (p=0.001). There was a tendency toward higher scores for "food responsiveness" (p=0.005) and "emotional overeating" (p=0.013) in participants with severe obesity. There were no differences in the scale of food neophobia. Overall, food neophobia positively correlated with lack of interest in food and negatively correlated with interest in food. Conclusions: The study showed significant differences in some domains of eating behavior among children and adolescents of the sample; however, no differences were found regarding food neophobia. These results may contribute to the improvement of future interventions related to infant eating behavior and food neophobia.

RESUMO Objetivo: Investigar e comparar o comportamento e a neofobia alimentares de crianças e adolescentes de diferentes faixas etárias, índice de massa corporal/idade e sexo. Métodos: Tratou-se de um estudo transversal, com amostra por conveniência, envolvendo 150 crianças e adolescentes com faixa etária entre 3 e 13 anos, de ambos os sexos, atendidas em um ambulatório pediátrico de um hospital escola em Uberaba, MG. Para avaliar o comportamento alimentar, utilizaram-se as subescalas do Child Eating Behaviour Questionnaire (CEBQ) e para a neofobia alimentar o Child Food Neophobia Scale (CFNS). Resultados: Verificaram-se pontuações mais altas nas subescalas "resposta à comida" (p=0,015), "prazer em comer" (p=0,002) e "sobreingestão emocional" (p=0,009) em crianças e adolescentes com idades maiores. As crianças com idades menores obtiveram pontuações mais altas nas subescalas como "resposta à saciedade" (p=0,004) e "ingestão lenta" (p=0,001). Observou-se uma tendência à maior pontuação quanto à "resposta à comida" (p=0,005) e à "sobreingestão emocional" (p=0,013) nos participantes com obesidade grave. Não houve diferenças quanto à escala de neofobia alimentar. De modo geral, a neofobia alimentar correlacionou-se positivamente com o desinteresse pela comida e negativamente com o interesse pela comida. Conclusões: O estudo indicou diferenças significantes em alguns domínios do comportamento alimentar entre as crianças e os adolescentes da amostra, porém não houve diferenças em relação à neofobia alimentar. Nesse sentido, esses resultados podem contribuir para o aprimoramento de intervenções futuras referentes ao comportamento e à neofobia alimentares infantis.

Perfil clínico e nutricional de crianças com alergia à proteína do leite de vaca / Clinical and nutritional profile of children with cow's milk protein allergy

RESUMO: Objetivo: Este estudo teve como objetivo identificar as principais características clínicas e nutricionais apresentadas pelas crianças com alergia à proteína do leite de vaca. Método: Tratou-se de um estudo observacional, transversal, quantitativo e correlacional, com amostra de conveniência, constituída por 22 crianças diagnosticados com alergia em diferentes estágios, na faixa etária de 6 meses a 6 anos, atendidas no ambulatório de Gastroenterologia Pediátrica do Hospital de Clínicas da Universidade Federal do Triângulo Mineiro. Resultados: Evidenciou-se eutrofia em 81,8% das crianças, risco de sobrepeso em 4,5%, sobrepeso em 9,1% e obesidade em 4,5%. Nenhum dos pacientes apresentou diagnóstico nutricional de magreza. Os sintomas gastrointestinais (diarreia, vômito e sangue vivo nas fezes) foram os que mais acometeram os pacientes avaliados. Todos os pacientes (100%) com alergia em remissão e alergia resolvida mostraram nível de ferritina sérica normal, porém, 14,3% das crianças em alergia ativa estavam com esse valor abaixo do recomendado para idade. As crianças apresentam desmame precoce do leite materno, alimentação complementar dentro do que é indicado e o uso de fórmulas infantis corretas para atingir as recomendações nutricionais para a idade. Conclusão: Os achados indicam que o perfil clínico e nutricional de crianças com alergia está em conformidade com o esperado no que tange à escolha das fórmulas e desenvolvimento das crianças. (AU)

ABSTRACT: Objective: this study aimed to identify the main clinical and nutritional characteristics presented by children with cow's milk protein allergy. Method: this was an observational, cross-sectional, quantitative and correlational study, with a convenience sample, consisting of 22 children diagnosed with allergy in different stages, aged between six months and six years, seen at the Pediatric Gastroenterology Outpatient Clinic of the Clinical Hospital of the Federal University of the Triângulo Mineiro. Results: eutrophy was evident in 81.8% of the children, overweight risk in 4.5%, overweight in 9.1%, and obesity in 4.5%. None of the patients presented a nutritional diagnosis of thinness. Gastrointestinal symptoms (diarrhea, vomiting, and bloody stools) were the most frequent in the evaluated patients. All patients (100%) with allergy in remission and allergy resolved showed a normal serum ferritin level, but 14.3% of the children in active allergy were below the recommended level for their age. The children had early weaning from breast milk, complementary feeding within what is indicated and the use of correct infant formulas to achieve the nutritional recommendations for age. Conclusion: the findings indicate that the clinical and nutritional profile of children with allergy is as expected with regard to the choice of formulas and the children's development. (AU)

Ângulo de Fase e Estado Nutricional de Crianças em Tratamento Antineoplásico / Phase Angle and Nutritional Status of Children in Antineoplastic Treatment / Ángulo de Fase y Estado Nutricional de los Niños Sometidos a Tratamiento Antineoplásico

Introdução: O câncer infantil gera um grande impacto para a saúde pública. A avaliação da composição corporal pela impedância bioelétrica fornece os valores de ângulo de fase, diretamente relacionados a alterações funcionais na membrana celular e que têm sido usados para avaliar o estado nutricional e o prognóstico. Objetivo: Avaliar a correlação entre o ângulo de fase e os parâmetros do estado nutricional de pacientes oncológicos pediátricos em tratamento. Método: Estudo transversal realizado em um hospital público universitário. O estado nutricional foi verificado por meio das medidas de peso, altura, circunferência do braço, dobra cutânea tricipital, índice de massa corporal e circunferência muscular do braço. O ângulo de fase foi determinado pela impedância bioelétrica. Resultados: Foram incluídos 13 pacientes com média de idade de 103,2±39,7 meses, sendo 61,5% do sexo masculino, com prevalência da leucemia entre os diagnósticos. Constatou-se que a maioria (53,8%) dos pacientes estava em eutrofia. Houve forte correlação do ângulo de fase com os parâmetros de massa magra (r=0,923; p=0,000), massa corporal celular (r=0,911; p=0,000), massa extracelular (r=0,897; p=0,000) e peso corporal (r=0,920; p=0,000). Conclusão: Observaram-se correlações expressivas entre o ângulo de fase e os indicadores antropométricos, reforçando a hipótese de que o ângulo de fase pode antecipar a identificação de alterações na composição corporal, possibilitando uma intervenção nutricional precoce e melhor prognóstico.

Introduction: Child cancer has a major impact on public health. The evaluation of body composition by bioelectric impedance provides the values of phase angle, causally related to functional changes in the cell membrane which have been used to evaluate nutritional status and prognosis. Objective: To evaluate the correlation between phase angle and parameters of the nutritional status of pediatric oncologic patients under treatment. Method: Cross-sectional study performed in a public university hospital. The nutritional status was evaluated by weight, height, arm circumference, triceps skin fold, body mass index and arm muscle circumference. The phase angle was determined by bioelectric impedance. Results: 13 patients were included with a mean age of 103.2±39.7 months, 61.5% males with prevalence of leukemia in the diagnoses. It was found that most (53.8%) of the patients were eutrophic. There was a strong correlation of the phase angle with the parameters of lean mass (r=0.923; p=0.000), cellular body mass (r=0.911; p=0.000), extracellular mass (r=0.897; p=0.000) and body weight (r=0.920; p=0.000). Conclusion: Expressive associations between phase angle and other anthropometric indicators were observed, reinforcing the hypothesis that it can anticipate the identification of changes in body composition, allowing early nutritional intervention and better prognosis.

Introducción: El cáncer infantil tiene un gran impacto en la salud pública. La evaluación de la composición corporal por impedancia bioeléctrica proporciona los valores del ángulo de fase, directamente relacionados con los cambios funcionales de la membrana celular y que se han utilizado para evaluar el estado nutricional y el pronóstico. Objetivo: Evaluar la correlación entre el ángulo de fase y los parámetros del estado nutricional de los pacientes oncológicos pediátricos en tratamiento. Método: Estudio transversal realizado en un hospital universitario público. El estado nutricional se evaluó por peso, altura, circunferencia del brazo, pliegue de la piel del tríceps, índice de masa corporal y circunferencia muscular del brazo. El ángulo de fase fue determinado por la impedancia bioeléctrica. Resultados: Se incluyeron 13 pacientes con una edad media de 103,2±39,7 meses, de los cuales el 61,5% eran hombres con prevalencia de leucemia entre los diagnósticos. Se descubrió que la mayoría (53,8%) de los pacientes eran eutróficos. Hubo una fuerte correlación del ángulo de fase con los parámetros de masa magra (r=0,923; p=0,000), masa corporal celular (r=0,911; p=0,000), masa extracelular (r=0,897; p=0,000) y peso corporal (r=0,920; p=0,000). Conclusión: Se observaron asociaciones expresivas entre el ángulo de fase y otros indicadores antropométricos, lo que refuerza la hipótesis de que puede anticiparse a la identificación de cambios en la composición corporal, lo que permite una intervención nutricional temprana y un mejor pronóstico.

ASSESSMENT OF EATING BEHAVIOR AND FOOD NEOPHOBIA IN CHILDREN AND ADOLESCENTS FROM UBERABA-MG.

OBJECTIVE: To investigate and compare the eating behavior and food neophobia of children and adolescents from different age groups, body mass index per age, and sex. METHODS: This was a cross-sectional study, with a convenience sample, involving 150 children and adolescents aged 3 to 13 years, of both sexes, treated at a pediatric outpatient clinic of a teaching hospital in the municipality of Uberaba-MG, Brazil. Subscales of the Child Eating Behavior Questionnaire (CEBQ) were used to evaluate eating behavior, and the Child Food Neophobia Scale (CFNS) was used to evaluate food neophobia. RESULTS: Higher scores were found in the subscales "food responsiveness" (p=0.015), "enjoyment of food" (p=0.002), and "emotional overeating" (p=0.009) among older children and adolescents. Younger children had higher scores in the subscales "satiety responsiveness" (p=0.004) and "slowness in eating" (p=0.001). There was a tendency toward higher scores for "food responsiveness" (p=0.005) and "emotional overeating" (p=0.013) in participants with severe obesity. There were no differences in the scale of food neophobia. Overall, food neophobia positively correlated with lack of interest in food and negatively correlated with interest in food. CONCLUSIONS: The study showed significant differences in some domains of eating behavior among children and adolescents of the sample; however, no differences were found regarding food neophobia. These results may contribute to the improvement of future interventions related to infant eating behavior and food neophobia.

Changes in lipid profiles of epileptic mouse model.

INTRODUCTION: Approximately 1% of the world's population is impacted by epilepsy, a chronic neurological disorder characterized by seizures. One-third of epileptic patients are resistant to AEDs, or have medically refractory epilepsy (MRE). One non-invasive treatment that exists for MRE includes the ketogenic diet, a high-fat, low-carbohydrate diet. Despite the KD's success in seizure attenuation, it has a few risks and its mechanisms remain poorly understood. The KD has been shown to improve metabolism and mitochondrial function in epileptic phenotypes. Potassium channels have implications in epileptic conditions as they have dual roles as metabolic sensors and control neuronal excitation. OBJECTIVES: The goal of this study was to explore changes in the lipidome in hippocampal and cortical tissue from Kv1.1-KO model of epilepsy. METHODS: FT-ICR/MS analysis was utilized to examine nonpolar metabolome of cortical and hippocampal tissue isolated from a Kv1.1 channel knockout mouse model of epilepsy (n = 5) and wild-type mice (n = 5). RESULTS: Distinct metabolic profiles were observed, significant (p < 0.05) features in hippocampus often being upregulated (FC ≥ 2) and the cortex being downregulated (FC ≤ 0.5). Pathway enrichment analysis shows lipid biosynthesis was affected. Partition ratio analysis revealed that the ratio of most metabolites tended to be increased in Kv1.1-/-. Metabolites in hippocampal tissue were commonly upregulated, suggesting seizure initiation in the hippocampus. Aberrant mitochondrial function is implicated by the upregulation of cardiolipin, a common component in the mitochondrial membrane. CONCLUSION: Generally, our study finds that the lipidome is changed in the hippocampus and cortex in response to Kv1.1-KO indicating changes in membrane structural integrity and synaptic transmission.