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The development of new approaches allowing for the early assessment of COVID-19 cases that are likely to become critical and the discovery of new therapeutic targets are urgently required. In this prospective cohort study, we performed proteomic and laboratory profiling of plasma from 163 COVID-19 patients admitted to Bauru State Hospital (Brazil) between 4 May 2020 and 4 July 2020. Plasma samples were collected upon admission for routine laboratory analyses and shotgun quantitative label-free proteomics. Based on the course of the disease, the patients were divided into three groups: (a) mild (n = 76) and (b) severe (n = 56) symptoms, whose patients were discharged without or with admission to an intensive care unit (ICU), respectively, and (c) critical (n = 31), a group consisting of patients who died after admission to an ICU. Based on our data, potential therapies for COVID-19 should target proteins involved in inflammation, the immune response and complement system, and blood coagulation. Other proteins that could potentially be employed in therapies against COVID-19 but that so far have not been associated with the disease are CD5L, VDBP, A1BG, C4BPA, PGLYRP2, SERPINC1, and APOH. Targeting these proteins' pathways might constitute potential new therapies or biomarkers of prognosis of the disease.

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Barium (Ba) is a non-essential element that can cause toxicity in living organisms and environmental contamination. Plants absorb barium predominantly in its divalent cationic form Ba2+. Sulfur (S) can decrease the availability of Ba2+ in the soil by causing its precipitation as barium sulfate, a compound known for its very low solubility. The objective of this study was to evaluate the effect of soil sulfate supply in soil Ba fractions, as well as on plant growth, and Ba and S uptake by lettuce plants grown in artificially Ba-contaminated soil under greenhouse conditions. The treatments consisted of five Ba doses (0, 150, 300, 450, and 600 mg kg-1 Ba, as barium chloride) combined with three S doses (0, 40, and 80 mg kg-1 S, as potassium sulfate). The treatments were applied to soil samples (2.5 kg) and placed in plastic pots for plant cultivation. The Ba fractions analyzed were extractable-Ba, organic matter-Ba, oxides associated-Ba, and residual-Ba. The results indicate that the extractable-Ba fraction was the main one responsible for Ba bioavailability and phytotoxicity, probably corresponding to the exchangeable Ba in the soil. The dose of 80 mg kg-1 of S reduced extractable-Ba by 30% at higher Ba doses while it increased the other fractions. Furthermore, S supply attenuated the growth inhibition in plants under Ba exposure. Thus, S supply protected the lettuce plants from Ba toxicity by reduction of Ba availability in soil and plant growth enhancement. The results suggest that sulfate supply is a suitable strategy for managing Ba-contaminated areas.

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The intestinal microbiota plays an important role in the immune response against viral infections, modulating both innate and adaptive immune responses. The cytokine storm is associated with COVID-19 severity, and the patient's immune status is influenced by the intestinal microbiota in a gut-lung bidirectional interaction. In this study, we evaluate the intestinal microbiota of Brazilian patients in different post-COVID-19 periods, and correlate this with clinical data and the antibiotic therapy used during the acute phase. DNA extracted from stool samples was sequenced and total anti-SARS-CoV-2 antibodies and C-reactive protein were quantified. Compared with controls, there were significant differences in the microbiota diversity in post-COVID-19 patients, suggesting an intestinal dysbiosis even several months after acute disease resolution. Additionally, we detected some genera possibly associated with the post-COVID-19 dysbiosis, including Desulfovibrio, Haemophillus, Dialister, and Prevotella, in addition to decreased beneficial microbes, associated with antibiotic-induced dysbiosis, such as Bifidobacterium and Akkermansia. Therefore, our hypothesis is that dysbiosis and the indiscriminate use of antibiotics during the pandemic may be associated with post-COVID-19 clinical manifestations. In our study, 39% (n = 58) of patients reported symptoms, including fatigue, dyspnea, myalgia, alopecia, anxiety, memory loss, and depression. These data suggest that microbiota modulation may represent a target for recovery from acute COVID-19 and a therapeutic approach for post-COVID-19 sequelae.

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BACKGROUND: Fluoride has become widely used in dentistry because of its effectiveness in caries control. However, evidence indicates that excessive intake interferes with the metabolic processes of different tissues. Thus, this study aimed to investigate the effects of long-term exposure to F on the parotid salivary gland of mice, from the analysis of oxidative, proteomic and genotoxic parameters. MATERIALS AND METHODS: The animals received deionized water containing 0, 10 or 50 mg/L of F, as sodium fluoride, for 60 days. After, parotid glands were collected for analysis of oxidative biochemistry, global proteomic profile, genotoxicity assessment and histopathological analyses. RESULTS: The results revealed that exposure to fluoride interfered in the biochemical homeostasis of the parotid gland, with increased levels of thiobarbituric acid reactive species and reduced glutathione in the exposed groups; as well as promoted alteration of the glandular proteomic profile in these groups, especially in structural proteins and proteins related to oxidative stress. However, genotoxic assessment demonstrated that exposure to fluoride did not interfere with DNA integrity in these concentrations and durations of exposure. Also, it was not observed histopathological alterations in parotid gland. CONCLUSIONS: Thus, our results suggest that long-term exposure to fluoride promoted modulation of the proteomic and biochemical profile in the parotid glands, without inducing damage to the genetic component. These findings reinforce the importance of rationalizing the use of fluorides to maximize their preventative effects while minimizing the environmental risks.

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The current treatment of leishmaniasis is based on a few drugs that present several drawbacks, such as high toxicity, difficult administration route, and low efficacy. These disadvantages raise the necessity to develop novel antileishmanial compounds allied with a comprehensive understanding of their mechanisms of action. Here, we elucidate the probable mechanism of action of the antileishmanial binuclear cyclopalladated complex [Pd(dmba)(µ-N3)]2 (CP2) in Leishmania amazonensis. CP2 causes oxidative stress in the parasite, resulting in disruption of mitochondrial Ca2+ homeostasis, cell cycle arrest at the S-phase, increasing the reactive oxygen species (ROS) production and overexpression of stress-related and cell detoxification proteins, and collapsing the Leishmania mitochondrial membrane potential, and promotes apoptotic-like features in promastigotes, leading to necrosis, or directs programmed cell death (PCD)-committed cells toward necrotic-like destruction. Moreover, CP2 reduces the parasite load in both liver and spleen in Leishmania infantum-infected hamsters when treated for 15 days with 1.5 mg/kg body weight/day CP2, expanding its potential application in addition to the already known effectiveness on cutaneous leishmaniasis for the treatment of visceral leishmaniasis, showing the broad spectrum of action of this cyclopalladated complex. The data presented here bring new insights into the CP2 molecular mechanisms of action, assisting the promotion of its rational modification to improve both safety and efficacy.

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OBJECTIVES: Salivary glands are affected during radiotherapy in the head and neck region, leading to a reduction in salivary flow and changes its composition. Besides negatively affecting the oral soft tissues, this can also lead to dental impairment. Thus, we evaluated the effect of radiotherapy in the proteomic profile of the saliva in patients with head and neck cancer (HNC). MATERIALS AND METHODS: HNC patients had their saliva collected before (BRT), during (2-5 weeks; DRT), and after (3-4 months; ART) radiotherapy. Saliva was also collected from healthy volunteers (control; C). Samples were processed for proteomic analysis. RESULTS: In total, 1055 proteins were identified, among which 46 were common to all groups, while 86, 86, 286, and 395 were exclusively found in C, BRT, DRT, and ART, respectively. Remarkably, alpha-enolase was increased 35-fold DRT compared with BRT, while proline-rich proteins were decreased. ART there was a 16-fold increase in scaffold attachment factor-B1 and a 3-fold decrease in alpha-enolase and several cystatins. When compared with C, salivary proteins of BRT patients showed increases cystatin-C, lysozyme C, histatin-1, and proline-rich proteins CONCLUSION/CLINICAL REVELANCE: Both HNC and radiotherapy remarkably change the salivary protein composition. Altogether, our results, for the first time, suggest investigating alpha-enolase levels in saliva DRT in future studies as a possible biomarker and strategy to predict the efficiency of the treatment. Moreover, our data provide important insights for designing dental products that are more effective for these patients and contribute to a better understanding of the progressive changes in salivary proteins induced by radiotherapy. Graphical abstract.

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Although fluoride (F) is well-known to prevent dental caries, changes in cell processes in different tissues have been associated with its excessive exposure. Thus, this study aimed to evaluate the effects of F exposure on biochemical, proteomic, and genotoxic parameters of submandibular glands. Twenty one old rats (n = 30) were allocated into three groups: 60 days administration of drinking water containing 10 mgF/L, 50 mgF/L, or only deionized water (control). The submandibular glands were collected for oxidative biochemistry, protein expression profile, and genotoxic potential analyses. The results showed that both F concentrations increased the levels of thiobarbituric acid-reactive substances (TBARS) and reduced glutathione (GSH) and changed the proteomic profile, mainly regarding the cytoskeleton and cellular activity. Only the exposure to 50 mgF/L induced significant changes in DNA integrity. These findings reinforce the importance of continuous monitoring of F concentration in drinking water and the need for strategies to minimize F intake from other sources to obtain maximum preventive/therapeutic effects and avoid potential adverse effects.

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Stimulation of saliva production is an alternative to improve the quality of life of patients treated by radiotherapy. However, there is no information about changes in the salivary proteome of stimulated and unstimulated saliva in these patients. OBJECTIVES: Thus, we evaluated the difference in the proteomic profile of stimulated and unstimulated saliva in patients with head and neck cancer (HNC) treated by radiotherapy. METHODS: Stimulated and unstimulated saliva were collected from 9 patients with HNC before (BRT), during (DRT; 2-5 weeks) and after (ART; 3-4 months) treatment. Healthy patients paired by age and gender also had their saliva collected (C; control group). The stimulated and unstimulated salivary flow were evaluated (p < 0.05). Salivary proteins were extracted and processed for shotgun proteomic analysis. RESULTS: Significant differences were observed between stimulated and unstimulated salivary flows for C and BRT (p greater than 0.001), but not for DRT and ART. Proteins involved with apoptosis, antibacterial and acid-resistance were decreased in stimulated saliva in comparison to unstimulated saliva DRT and ART. Isoforms of keratins were not identified in control and BRT. CONCLUSION: there is a marked difference in the protein profile of stimulated and unstimulated salivary flows in HNC patients treated by radiotherapy. In addition, saliva stimulation in patients with HNC decreases important proteins involved with dental protection. The unstimulated salivary flow seems to be the best alternative to search for biomarkers. Our results contribute in an unprecedented way to understand the changes in the salivary proteome of different flows in HNC patients undergoing radiotherapy treatment.

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Dysbiosis, associated with barrier disruption and altered gut-brain communications, has been associated with multiple sclerosis (MS). In this study, we evaluated the gut microbiota in relapsing-remitting patients (RRMS) receiving disease-modifying therapies (DMTs) and correlated these data with diet, cytokines levels, and zonulin concentrations. Stool samples were used for 16S sequencing and real-time PCR. Serum was used for cytokine determination by flow cytometry, and zonulin quantification by ELISA. Pearson's chi-square, Mann-Whitney, and Spearman's correlation were used for statistical analyses. We detected differences in dietary habits, as well as in the gut microbiota in RRMS patients, with predominance of Akkermansia muciniphila and Bacteroides vulgatus and decreased Bifidobacterium. Interleukin-6 concentrations were decreased in treated patients, and we detected an increased intestinal permeability in RRMS patients when compared with controls. We conclude that diet plays an important role in the composition of the gut microbiota, and intestinal dysbiosis, detected in RRMS patients could be involved in increased intestinal permeability and affect the clinical response to DTMs. The future goal is to predict therapeutic responses based on individual microbiome analyses (personalized medicine) and propose dietary interventions and the use of probiotics or other microbiota modulators as adjuvant therapy to enhance the therapeutic efficacy of DMTs.

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The enteric nervous system is responsible for controlling the gastrointestinal tract (GIT) functions. Enteric neuropathies are highly correlated to the development of several intestinal disturbances. Fluoride (F) is extensively applied for dental health improvement and its ingestion can promote systemic toxicity with mild to severe GIT symptomatology and neurotoxicity. Although F harmful effects have been published, there is no information regarding noxiousness of a high acute F exposure (25 mg F/kg) on enteric neurons and levels of expression of intestinal proteins in the duodenum. Quantitative proteomics of the duodenum wall associated to morphometric and quantitative analysis of enteric neurons displayed F effects of a high acute exposure. F-induced myenteric neuroplasticity was characterized by a decrease in the density of nitrergic neurons and morphometric alterations in the general populations of neurons, nitrergic neurons, and substance P varicosities. Proteomics demonstrated F-induced alterations in levels of expression of 356 proteins correlated to striated muscle cell differentiation; generation of precursor metabolites and energy; NADH and glutathione metabolic process and purine ribonucleoside triphosphate biosynthesis. The neurochemical role of several intestinal proteins was discussed specially related to the modulation of enteric neuroplasticity. The results provide a new perspective on cell signaling pathways of gastrointestinal symptomatology promoted by acute F toxicity.

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Fluoride (F) is largely employed in dentistry, in therapeutic doses, to control caries. However, excessive intake may lead to adverse effects in the body. Since F is absorbed mostly from the gastrointestinal tract (GIT), gastrointestinal symptoms are the first signs following acute F exposure. Nevertheless, little is known about the mechanistic events that lead to these symptoms. Therefore, the present study evaluated changes in the proteomic profile as well as morphological changes in the jejunum and ileum of rats upon acute exposure to F. Male rats received, by gastric gavage, a single dose of F containing 0 (control) or 25 mg/Kg for 30 days. Upon exposure to F, there was a decrease in the thickness of the tunic muscularis for both segments and a decrease in the thickness of the wall only for the ileum. In addition, a decrease in the density of HuC/D-IR neurons and nNOS-IR neurons was found for the jejunum, but for the ileum only nNOS-IR neurons were decreased upon F exposure. Moreover, SP-IR varicosities were increased in both segments, while VIP-IR varicosities were increased in the jejunum and decreased in the ileum. As for the proteomic analysis, the proteins with altered expression were mostly negatively regulated and associated mainly with protein synthesis and energy metabolism. Proteomics also revealed alterations in proteins involved in oxidative/antioxidant defense, apoptosis and as well as in cytoskeletal proteins. Our results, when analyzed together, suggest that the gastrointestinal symptoms found in cases of acute F exposure might be related to the morphological alterations in the gut (decrease in the thickness of the tunica muscularis) that, at the molecular level, can be explained by alterations in the gut vipergic innervation and in proteins that regulate the cytoskeleton.

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Lead (Pb) is an environmental and occupational neurotoxicant after long-term exposure. This study aimed to investigate the effects of systemic Pb exposure in rats from adolescence to adulthood, evaluating molecular, morphologic and functional aspects of hippocampus. For this, male Wistar rats were exposed to 50 mg/kg of Pb acetate or distilled water for 55 days by intragastric gavage. For the evaluation of short-term and long-term memories, object recognition and step-down inhibitory avoidance tests were performed. At the end of the behavioral tests, the animals were euthanized and the hippocampus dissected and processed to the evaluation of: Pb content levels in hippocampal parenchyma; Trolox equivalent antioxidant capacity (TEAC), glutathione (GSH) and malondialdehyde (MDA) levels as parameters of oxidative stress and antioxidant status; global proteomic profile and neuronal degeneration by anti-NeuN immunohistochemistry analysis. Our results show the increase of Pb levels in the hippocampus of adult rats exposed from adolescence, increased MDA and GSH levels, modulation of proteins related to neural structure and physiology and reduced density of neurons, hence a poor cognitive performance on short and long-term memories. Then, the long-term exposure to Pb in this period of life may impair several biologic organizational levels of the hippocampal structure associated with functional damages.

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Water-soluble phosphate fertilizers release phosphorus (P) to soils promptly, causing P fixation and low plant availability in highly weathered tropical soils. Therefore, the development of strategies to improve P use efficiency is needed. We hypothesized that biochar-based fertilizers (BBFs) can provide available P to plants and improve P use efficiency when compared with soluble fertilizers. Thus, triple superphosphate (TSP) and phosphoric acid (H3PO4) were pyrolyzed with and without magnesium oxide (MgO) and poultry litter to produce slow-release P BBFs. A pot experiment under greenhouse conditions was performed to evaluate agronomic efficiency of BBFs compared with TSP in an Oxisol. The treatments were incubated over 100 days after the application of 25, 50, 100, and 200 mg kg-1 of P. Three controls were used, including 200 mg kg-1 of P as TSP incubated for 100 days (named TSPincubation) and applied immediately before sowing (named TSPplanting) and a negative control (without P). Marandu grass (Urochloa brizantha cv. Marandu) was cultivated in pots for three cycles of 40 days each. After cultivation, a sequential extraction procedure was used to determine the P distribution among different P pools. The shoot dry matter yield in the first cropping cycle was higher at the highest P rate for TSPplanting. PLB-H3PO4-MgO showed 9% increase in the shoot dry matter when compared with TSPincubation in the first cropping cycle. In subsequent cropping cycles, all BBFs promoted higher biomass yield when compared with TSPplanting. There was an increase in the labile and moderately labile P fractions in soil after cultivation with PLB-TSP. The results suggest that BBFs can enhance P use efficiency in tropical soils in the middle- to long-term run due to slow-release profile that prevent P fixation and promote higher residual effect of fertilization.

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The development of metallomics techniques has allowed for metallomics analysis of biological systems, enabling a better understanding of the response mechanisms for different stimuli, their relationship to metallic species, and the characterization of biomarkers. In this study, a metallomics analysis of the muscle tissue of Nile tilapia was used to aid the understanding of the molecular mechanisms involved in zinc absorption in this fish species when fed organic and/or inorganic sources of zinc and to identify possible biomarkers for the absorption of this micromineral. To accomplish this, the fish were separated into three groups of 24 g, 74 g, and 85 g initial weights, and each group, respectively, was fed a zinc-free diet (control group, G1), a diet containing zinc found in organic sources (treatment 1, G2), and a diet containing zinc from an inorganic source (treatment 2, G3). Two-dimensional polyacrylamide (2D PAGE) gel electrophoresis was used to separate the proteins of the muscle tissue. Subsequently, the expression profiles of protein spots in the samples where zinc was applied in different concentrations were compared, using the software ImageMaster 2D Platinum version 7.0, to identify proteins that were differentially expressed. The identified proteins were then exposed to atomic absorption spectrometry in a graphite furnace to determine zinc mapping and were subsequently characterized via electrospray ionization tandem mass spectrometry (ESI-MS/MS). The metallomic analysis identified 15 proteins differentially expressed and associated with zinc, leading to the conclusion that three metal-binding proteins presented as possible biomarkers of zinc absorption in fish.

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The high concentrations of mercury found in Amazon have been intensively studied by the scientific community in the last decades. These mercurial species bind preferentially to proteins. Therefore, this work proposal sought to obtain the fractionation, identification and study of mercury - bound proteins present in samples of muscular and hepatic tissue from fish collected in the reservoir of the Jirau Hydroelectric Power Plant - on the Madeira River. Two-dimensional electrophoresis (2D-PAGE) for protein fractionation, graphite furnace atomic absorption spectrometry (GFAAS) for the quantification of mercury and Mass Spectrometry (ESI-MS/MS) were used for the identification of proteins. Concluding the work with analysis of graphics from the Blast2go program. Two mercury - bound proteins were identified as triosephosphate isomerase A and Protein FAM45A. The data generated by the bioinformatics programs confirm the tendency of these proteins to be linked to mercury and elucity the possibles existing physiological and cellular interactions.

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Pythium insidiosum is the etiologic agent of pythiosis, a life-threatening disease that affects human and animals, has difficult diagnosis, and therapy. Studies on protein characterization of P. insidiosum are scarce, so we aimed to determine the protein profile of P. insidiosum by mass spectrometry and bioinformatics strategies targeting in proteins that may act as putative virulence factors. Therefore, an extraction protocol was standardized to obtain the total proteins of P. insidiosum. By the analysis of Image Master 2D Platinum software, it was found that 186 spots ranging between 12 and 89 KDa and isoelectric point from 4 to 7. By the analysis of 2D-SDS-PAGE it was possible to visualize and excise 103 spots, which were hydrolyzed with trypsin and submitted to mass spectrometry, resulting in the identification of 36 different proteins. Three of them were classified as proteins supposedly related to virulence factors due to its functions, such as glucan 1,3-beta glucosidase, Heat shock protein (Hsp) 70 and enolase. These results may contribute to a better understanding of the virulence factors of this medically important oomycete, as well as to subsidize new studies on diagnosis and therapeutic approaches.

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Genes expressed during amelogenesis are candidates to increase the risk of dental fluorosis (DF). Thus, this study aimed to evaluate the association between polymorphisms in enamel development genes and susceptibility to DF in mice. Mice of both sexes, representing strains 129P3/J (n = 20; resistant to DF) and A/J (n = 20; susceptible to DF), were divided into 2 groups. Each strain received a diet with a low concentration of fluoride (F) and drinking water containing 0 or 50 mg/L of F for 6 weeks. Clinical evaluation and analysis of Vickers enamel microhardness of the incisors were performed. Livers were collected for genomic DNA extraction. Seventeen genetic polymorphisms in Amelx, Ambn, Ambn, Col14a1, Col1a1, Col5a2, Enam, Fam20a, Fam83h, Foxo1, Klk4, Mmp20, Serpinf1, Serpinh1, Smad3, Tuft1, and Wdr72 were genotyped by real-time PCR using Taqman chemistry. Overrepresentation of alleles and genotypes in DF was evaluated using the χ2 test with an alpha of 5%. The clinical aspects of the enamel and the surface enamel microhardness confirmed the DF condition. In the polymorphisms rs29569969, rs13482592, and rs13480057 in Ambn, Col14a1, and Mmp20, respectively, genotype and allele distributions were statistically significantly different between A/J and 129P3/J strains (p < 0.05). In conclusion, polymorphisms in Ambn, Col14a1, and Mmp20 are associated with the susceptibility to DF.

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ABSTRACT Objective: to verify the correlation between vital capacity and maximum phonation times of /ė/ (unvoiced) and /s/, as well as compare and relate them with the professional voice use and age in women with functional or organic-functional dysphonia. Methods: a retrospective research with 524 records of dysphonic patients from a school clinic, including young adult women with a speech-language diagnosis of functional or organic-functional dysphonia based on medical reports. Neurological and psychiatric alterations, previous speech therapy treatment, symptoms of flu or allergies on the day of evaluation, pulmonary disease, organic dysphonia diagnosis, and hearing loss, were excluded. The sample resulted in 14 women with functional dysphonia and 21 with organic-functional dysphonia. Data on professional voice use, as well as results for vital capacity and maximum phonation times were collected. The data were statistically analyzed at a 5% significance level. Results: There was a positive correlation for both groups of dysphonic patients between the maximum phonation times of /ė/ and of /s/, as well as the maximum phonation times of /ė/, /s/, and vital capacity. Higher values for vital capacity and maximum times of /s/ and /ė/ for voice professionals were seen. The maximum phonation times of /ė/ were lower than those of /s/. Conclusion: as the maximum phonation times of /ė/ increased, the maximum phonation times of /s/ and the vital capacity also augmented in both groups, demonstrating the interrelation among these variables; there was no relation with the other variables studied.

RESUMO Objetivo: verificar a correlação entre capacidade vital, tempos máximos de fonação de /ė/ (sem sonorização) e /s/ e comparar e relacionar com o uso profissional da voz e a idade em mulheres com disfonia funcional ou organofuncional. Métodos: pesquisa retrospectiva com 524 registros de disfônicos de uma clínica-escola. Foram incluídas: mulheres adultas jovens, diagnóstico fonoaudiológico de disfonia funcional ou organofuncional, com base no laudo médico. Foram excluídas: alterações neurológicas, psiquiátricas, tratamento fonoaudiológico prévio, quadro gripal ou alérgico no dia das avaliações, doença pulmonar, diagnóstico de disfonia orgânica, perda auditiva. A amostra resultou em 14 mulheres com disfonia funcional e 21 com disfonia organofuncional. Coletaram-se dados sobre e uso profissional da voz e resultados da capacidade vital e dos tempos máximos de fonação. Os dados foram analisados estatisticamente com nível de significância de 5%. Resultados: correlação positiva para ambos os grupos de disfônicas entre: tempo máximo de fonação de /ė/ e tempo máximo de fonação de /s/; e entre tempo máximo de fonação de /ė/, /s/ e capacidade vital. Maiores valores de capacidade vital, tempo máximo de /s/ e /ė/ nas profissionais da voz. Os valores de tempo máximo de fonação de /ė/ foram menores do que /s/. Conclusão: à medida que o tempo máximo de fonação de /ė/ aumentou, o tempo máximo de fonação de /s/ e a capacidade vital também aumentaram em ambos os grupos, evidenciando a inter-relação entre essas variáveis; não havendo relação com as demais variáveis estudadas.

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Gastrointestinal symptoms are the first signs of fluoride (F) toxicity. In the present study, the jejunum of rats chronically exposed to F was evaluated by proteomics, as well as by morphological analysis. Wistar rats received water containing 0, 10 or 50 mgF/L during 30 days. HuC/D, neuronal Nitric Oxide (nNOS), Vasoactive Intestinal Peptide (VIP), Calcitonin Gene Related Peptide (CGRP), and Substance P (SP) were detected in the myenteric plexus of the jejunum by immunofluorescence. The density of nNOS-IR neurons was significantly decreased (compared to both control and 10 mgF/L groups), while the VIP-IR varicosities were significantly increased (compared to control) in the group treated with the highest F concentration. Significant morphological changes were seen observed in the density of HUC/D-IR neurons and in the area of SP-IR varicosities for F-treated groups compared to control. Changes in the abundance of various proteins correlated with relevant biological processes, such as protein synthesis, glucose homeostasis and energy metabolism were revealed by proteomics.

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This study detected changes in the protein profile of the acquired enamel pellicle (AEP) formed in vivo after rinsing with whole milk, fat-free milk, or water. Nine subjects in good oral condition took part in the study. The acquired pellicle was formed in the morning, for 120 min, after prophylaxis with pumice. Following this, the volunteers rinsed with 10 mL of whole milk, fat-free milk, or deionized water for 30 s, following a blinded crossover protocol. After 60 min, the pellicle was collected with filter paper soaked in 3% citric acid and processed for analysis by liquid chromatography-electrospray ionization tandem mass spectrometry. The obtained tandem mass spectrometry spectra were searched against a human protein database (Swiss-Prot). The proteomic data related to protein quantification were analysed using the PLGS software. A total of 260 proteins were successfully identified in the AEP samples collected from all groups. Forty-nine were common to all 3 groups, while 72, 62, and 49 were specific to the groups rinsing with whole milk, fat-free milk, and water, respectively. Some were typical components of the AEP, such as cystatin-B, cystatin-SN, isoforms of α-amylase, IgA and IgG, lysozyme C, protein S100 A78, histatin-1, proline-rich protein 27, statherin, and lactotransferrin. Other proteins are not commonly described as part of the AEP but could act in defence of the organism against pathogens. Distinct proteomic profiles were found in the AEP after rinsing with whole or fat-free milk, which could have an impact on bacterial adhesion and tooth dissolution. The use of fat-free milk could favourably modulate the adhesion of bacteria to the AEP as well as biofilm formation when compared with whole milk.

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