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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

The objective of the study was to evaluate the effect of propolis on the digestibility of minerals in the diet of blue-fronted amazon (Amazona aestiva). For this, 10 adult birds (5 males and 5 females) were separated into two groups (Control and Propolis 0.5% m/m). The apparent digestibility coefficient (ADC) of the following minerals was analyzed by spectrometry: calcium (Ca), phosphorus (P), iron (Fe), magnesium (Mg), sodium (Na), potassium (K), zinc (Zn), manganese (Mn) and nickel (Ni). ANOVA was used to evaluate ADC analyzes of the minerals, as well as the Pearson correlation between the ADC of the control and propolis rations. There were lower correlations in the ADC of the minerals of the birds that received propolis ration. It was concluded that parrots consuming propolis-containing ration showed better ADC levels, possibly due to the reduction in the interaction between the minerals and the alteration of the intestinal flora by propolis.

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

Made available in DSpace on 2021-06-25T11:13:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 In this study, we analyzed Apis mellifera royal jelly proteome alterations induced by supplementation with different concentrations of dietary organic zinc (0, 25, 50, and 75 ppm). Proteins extracted from royal jelly were separated by electrophoresis and isoelectric focusing, and zinc-containing protein spots were identified by tandem mass spectrometry. Organic zinc-containing diets modified total royal jelly protein content and altered the expression of five major royal jelly proteins (MRJP1, MRJP2, MRJP3, MRJP5, and MRJP8). While the diet with the lowest tested zinc content (Zn25) increased total royal jelly protein content and expression of MRJP1, diets with higher zinc contents (Zn50 and Zn75) decreased MRJP expression. Our results demonstrate that important proteins and metabolic processes are modified in royal jelly produced by honey bees supplemented with different concentrations of organic zinc source, affecting nutrition and maintenance of colonies. Center of Education Science and Technology in Rational Beekeeping (NECTAR) College of Veterinary Medicine and Animal Sciences UNESP–São Paulo State University Institute of Biosciences Department of Biochemistry UNESP - São Paulo State University Center of Education Science and Technology in Rational Beekeeping (NECTAR) College of Veterinary Medicine and Animal Sciences UNESP–São Paulo State University Institute of Biosciences Department of Biochemistry UNESP - São Paulo State University

Made available in DSpace on 2020-12-12T02:08:39Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-01-01 This manuscript describes the results of a metalloproteomic study of mercury in samples of muscle and liver tissue of the species Serrasalmus rhombeus, popularly known as black piranha and characterised as the most voracious and aggressive predator in the Brazilian Amazon. The metalloproteomic study involved using two-dimensional electrophoresis (2D PAGE) to fractionate the proteome of the muscle and liver tissue samples, along with atomic absorption spectrometry in a graphite furnace (GFAAS) to identify mercury associated with protein SPOTs and mass spectrometry with electrospray ionisation (ESI-MS/MS) to characterise the mercury-binding proteins. The protein SPOTs characterised showed concentrations in the order of 156 mg kg−1, which ranks as the highest concentrations of mercury determined so far in metalloproteomic studies involving fish species in the Amazon region. Based on FASTA sequences of proteins characterised by ESI-MS/MS, bioinformatics studies were performed that allowed identifying nine proteins with characteristics of biomarkers of mercury exposure. Of those proteins, glutathione peroxidase stands out as an enzyme of great importance in the antioxidant defence of organisms subjected to oxidative stress caused by xenobiotics. School of Veterinary Medicine and Animal Science São Paulo State University (UNESP), Rua Prof R. Dr. Valter Maurício Corrêa, s/n Venom and Venomous Animals Study Center (CEVAP) of São Paulo State University (UNESP), Rua José Barbosa de Barros, 1780 - Vila Ema Institute of Biosciences São Paulo State University (UNESP), Rua Prof. Dr. Antonio Celso Wagner Zanin, 250 Faculty of Science and Engineering São Paulo State University (UNESP), R. Domingos da Costa Lopes, 780 - Jardim Itaipu School of Veterinary Medicine and Animal Science São Paulo State University (UNESP), Rua Prof R. Dr. Valter Maurício Corrêa, s/n Venom and Venomous Animals Study Center (CEVAP) of São Paulo State University (UNESP), Rua José Barbosa de Barros, 1780 - Vila Ema Institute of Biosciences São Paulo State University (UNESP), Rua Prof. Dr. Antonio Celso Wagner Zanin, 250 Faculty of Science and Engineering São Paulo State University (UNESP), R. Domingos da Costa Lopes, 780 - Jardim Itaipu

Made available in DSpace on 2022-04-28T19:44:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-04-01 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. Graphical abstract: [Figure not available: see fulltext.] Institute of Biosciences São Paulo State University (UNESP), São Paulo University of Nebraska (UNL) College of Planaltina University of Brasília (UNB), Distrito Federal University of São Paulo (USP), São Paulo Institute of Biosciences São Paulo State University (UNESP), São Paulo

Made available in DSpace on 2021-06-25T11:02:27Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-06-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Pythiosis, whose etiological agent is the oomycete Pythium insidiosum, is a life-threatening disease that occurs mainly in tropical and subtropical countries, affecting several animal species. It is frequently found in horses in Brazil and humans in Thailand. The disease is difficult to diagnose because the pathogen’s hyphae are often misdiagnosed as mucoromycete fungi in histological sections. Additionally, there is no specific antigen to use for rapid diagnosis, the availability of which could improve the prognosis in different animal species. In this scenario, we investigated which P. insidiosum antigens are recognized by circulating antibodies in horses and humans with pythiosis from Brazil and Thailand, respectively, using 2D immunoblotting followed by mass spectrometry for the identification of antigens. We identified 23 protein spots, 14 recognized by pooled serum from horses and humans. Seven antigens were commonly recognized by both species, such as the heat-shock cognate 70 KDa protein, the heat-shock 70 KDa protein, glucan 1,3-beta-glucosidase, fructose-bisphosphate aldolase, serine/threonine-protein phosphatase, aconitate hydratase, and 14-3-3 protein epsilon. These results demonstrate that there are common antigens recognized by the immune responses of horses and humans, and these antigens may be studied as biomarkers for improving diagnosis and treatment. Department of Chemical and Biological Sciences Institute of Biosciences São Paulo State University (UNESP) Department of Pathology Faculty of Medicine Ramathibodi Hospital Mahidol University Faculty of Veterinary Medicine and Animal Science (FMVZ) São Paulo State University (UNESP) Department of Biological Sciences Bauru School of Dentistry University of São Paulo (USP) Department of Microbiology Immunology and Parasitology Cell Biology Division Federal University of São Paulo (UNIFESP) Center for the Study of Venoms and Venomous Animals (CEVAP) São Paulo State University (UNESP) Graduate Program in Tropical Diseases Botucatu Medical School (FMB) São Paulo State University (UNESP) Department of Chemical and Biological Sciences Institute of Biosciences São Paulo State University (UNESP) Faculty of Veterinary Medicine and Animal Science (FMVZ) São Paulo State University (UNESP) Center for the Study of Venoms and Venomous Animals (CEVAP) São Paulo State University (UNESP) Graduate Program in Tropical Diseases Botucatu Medical School (FMB) São Paulo State University (UNESP) CAPES: 001 FAPESP: 2016/10804-8 FAPESP: 2018/08009-0 FAPESP: 2019/08761-7

Made available in DSpace on 2019-10-06T16:27:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-06-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Protocols that improve growth performance in fish while assuring product quality are important for aquaculture. Fasting followed by refeeding may promote compensatory growth, thus optimizing growth performance. During fasting and refeeding, fast-twitch muscle, which comprises most of fish fillet, undergoes intense plasticity. In this work, we studied the proteome of pacu (Piaractus mesopotamicus)fast-twitch muscle after 30 days of fasting (D30), 30 days of refeeding (D60)and 60 days of refeeding (D90)with two-dimensional electrophoresis, mass spectrometry and bioinformatics. Body mass, growth rate and muscle histology were also assessed. At D30, fish presented muscle catabolism and decreased growth. Proteomic analysis showed that metabolism proteins were the most affected, up and downregulated. Cytoskeleton and amino acid biosynthesis proteins were downregulated, while nuclear and regulatory proteins were upregulated. At D60, fish showed accelerated growth, despite the body mass not completely recovering. Metabolism proteins were still the most affected. Amino acid biosynthesis proteins became upregulated, while cytoskeleton proteins remained downregulated. At D90, the fish presented total compensatory growth. Many metabolic proteins were up or downregulated. Few cytoskeleton proteins remained differentially expressed. Amino acid biosynthesis proteins were mostly upregulated, but less than at D60. Prolonged fasting followed by refeeding also led to the regulation of possible meat quality biomarkers, such as antioxidant enzymes. This fact suggests possible consequences of this protocol on fish meat quality. Our work also enriches our knowledge on proteomic changes during muscle plasticity that occur during fasting and refeeding diet protocols. Department of Morphology Institute of Biosciences of Botucatu São Paulo State University (UNESP) Department of Chemistry and Biochemistry Institute of Biosciences of Botucatu São Paulo State University (UNESP) Department of Biology University of Western São Paulo (UNOESTE) São Paulo Agency for Agribusiness Technology (APTA) Department of Morphology Institute of Biosciences of Botucatu São Paulo State University (UNESP) Department of Chemistry and Biochemistry Institute of Biosciences of Botucatu São Paulo State University (UNESP) CNPq: 116795/2016 FAPESP: 2017/12237-6 CNPq: 302656/2015-4 CNPq: 447233/2014

Bioactive peptides can provide health benefits due to different mechanisms. The aims of the present study are to produce bioactive peptides from bovine and goat milk subjected to the proteolytic activity of Aspergillus oryzae and Aspergillus flavipes enzymes, as well as to assess their putative antimicrobial and antioxidant activity. Bioactive peptides were successfully generated from proteases of fungi cultivated in solid-state fermentation. The generated peptides were effective against all tested bacteria and fungi. There was antioxidant activity, up to 92.5% DPPH reduction and ORAC stabilization at 52.5 μmol μL−1 of Trolox Equivalent. The generation of milk-specific sequences peptides in the samples was obtained through 2D-PAGE fractioning followed by mass spectrometry (ESI-MS/MS). Based on results in the present study, milk bioactive peptides presenting broad antimicrobial action and antioxidant activity spectra can be cost-effectively produced through solid-state fermentation. The herein addressed approach can be valuable for the pharmaceutical and food industries.

Mercury has the ability to bind to a variety of biomolecules, which can compromise its structure and functionality and thus promote its toxic effects. The aim of this study is to identify possible mercury biomarkers in muscle samples of Plagioscion squamosissimus (carnivorous fish) and Colossoma macropomum (omnivorous fish), from the Amazon region. The muscle proteome of fish species was separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and the total mercury concentrations in protein spots were determined by graphite furnace atomic absorption spectrometry (GFAAS). The protein spots containing mercury were characterized by electrospray ionization tandem mass spectrometry (ESI-MS/MS). The mercury concentrations in the protein spots were in the range of 1.10 ± 0.02–23.90 ± 0.33 μg g−1. The proteins phosphoglycerate mutase 2 (P. squamosissimus), hemoglobin β and cytochrome P450scc (C. macropomum), identified by ESI-MS/MS and showing the highest values of mercury concentration, may be considered possible mercury biomarkers.

Shear stress changes are associated with a repertory of signaling cascade modulating vascular phenotype. As shear stress-related tensional forces might be associated with pathophysiological susceptibility, a more comprehensive molecular map needs to be addressed. Thus, we subjected human umbilical vein endothelial cells (HUVECs) to a circuit of different tensional forces in vitro considering the following three groups: (a) physiological blood flow shear stress condition (named Normo), (b) a hypertensive blood flow shear stress (named Hyper), and (c) these hyper-stressed cells were returned to Normo condition (named Return). The samples were properly collected to allow different methodologies analysis. Our data showed a pivotal involvement of c-Src on driving the mechanotransduction cascade by modulating signaling related with adhesion, survival (PI3K/Akt) and proliferative phenotype. Moreover, c-Src seems to develop important role during extracellular matrix remodeling. Additionally, proteomic analysis showed strong involvement of heat shock protein 70 (HSP70) in the hypertensive-stressed cells; it being significantly decreased in return phenotype. This result prompted us to investigate 20S proteasome as an intracellular proteolytic alternative route to promote the turnover of those proteins. Surprisingly, our data reveled significant overexpression of sets of proteasome subunit α-type (PSMA) and β-type (PSMB) genes. In conjunction, our data showed c-Src as a pivotal protein to drive mechanotransduction in endothelial cells in a HSP70-dependent turnover scenario. Because shear patterns is associated with pathophysiological changes, such as atherosclerosis and hypertension, these results paved new road to understand the molecular mechanism on driving mechanotransduction in endothelial cells and, if drugable, these targets must be considered within pharmacological treatment optimization.

Background Diets for feedlot cattle require higher energy density, thus contributing to the high rate of fermentable carbohydrate. The use of feed additives is necessary to reduce possible metabolic disorders. The objective of this study was to analyze the post-rumen effects of different levels of starch (25, 35, and 45%) and additives (Monensin, Blend of essential oil + exogenous α-Amylase) in diets for Nellore cattle feedlot. The cecum tissue proteome was separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and then, with the verification of differentially expressed protein SPOTS, these were characterized by electrospray ionization mass spectrometry (ESI-MS/MS).Results The expression of nine enzymes participating in the Steps of the glycolysis pathway was verified, such as: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Glyceraldehyde-3-phosphate dehydrogenase like-17 protein, Triosephosphate isomerase (Step 1); Phosphoglycerate mutase and Phosphoglycerate mutase 2 (Step 2); Alpha-enolase (ENO1), Beta-enolase (ENO3) and Fructose-bisphosphate aldolase (ALDOB) (Step 4); and Pyruvate Kinase (PKM) (Step 5). There was expression of three enzymes linked to catalytic activities participating in the synthesis of lactate from pyruvate: L-lactate dehydrogenase B, L-lactate dehydrogenase A chain and L-lactate dehydrogenase. The ATP synthase subunit beta and ATP synthase subunit beta_mitochondrial participate in the electron transport chain, producing ATP from ADP in the presence of a proton gradient across the membrane. Due to the manipulation of diets, the expression of the Leukocyte elastase inhibitor protein, associated with the inflammatory response.Conclusions The use of blends of essential oil associated with α-amylase as a feed additive promoted the greater expression of enzymes in the pathway of glycolysis and gluconeogenesis (and the absence of proteins linked to inflammation (Leukocyte elastase inhibitor) in cecum tissues. On the other hand, the increase in starch in the diets promoted a reduction in enzymes linked to carbohydrate degradation with increased responses linked to inflammatory injuries.

Made available in DSpace on 2021-06-25T11:06:54Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-01-01 The developmental origins of health and disease concept links adult diseases with early-life exposure to inappropriate environmental conditions. Intrauterine and postnatal malnutrition may lead to an increased incidence of type 2 diabetes, obesity, and cardiovascular diseases. Maternal malnutrition (MM) has also been associated with prostate carcinogenesis. However, the molecular mechanisms associated with this condition remain poorly understood. Using a proteomic analysis, we demonstrated that MM changed the levels of proteins associated with growth factors, estrogen signaling, detoxification, and energy metabolism in the prostate of both young and old rats. These animals also showed increased levels of molecular markers of endoplasmic reticulum function and histones. We further performed an in silico analysis that identified commonly deregulated proteins in the ventral prostate of old rats submitted to MM with a mouse model and patients with prostate cancer. In conclusion, our results demonstrated that estrogenic signaling pathways, endoplasmic reticulum functions, energy metabolism, and molecular sensors of protein folding and Ca2+ homeostasis, besides histone, and RAS-GTPase family appear to be involved in this process. Knowledge of these factors may raise discussions regarding the role of maternal dietary intervention as a public policy for the lifelong prevention of chronic diseases. Department of Structural and Functional Biology Institute of Biosciences Sao Paulo State University (UNESP) Department of Chemical and Biological Sciences Institute of Biosciences Sao Paulo State University (UNESP) Department of Structural and Functional Biology Institute of Biosciences Sao Paulo State University (UNESP) Department of Chemical and Biological Sciences Institute of Biosciences Sao Paulo State University (UNESP)

Made available in DSpace on 2020-12-12T01:11:35Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-10-01 Recent studies have demonstrated the association of mercury (Hg) with some fish proteins, milk, and hair from individuals exposed to the element in the Amazon. However, few studies involve identifying biomarkers of mercury exposure. Therefore, the present study aimed to identify potential biomarkers of Hg exposure in fish. For this, the muscular tissues of two species of fish (Prochilodus lineatus and Mylossoma duriventre) that feed the Amazonian human population were analyzed. Through the analyses obtained by graphite furnace atomic absorption spectrometry (GFAAS), it was possible to identify four protein SPOTS where mercury was present. These SPOTS, identified by mass spectrometry (ESI-MS/MS), included parvalbumin and ubiquitin-40S ribosomal protein S27a, and these being metalloproteins with biomarker characteristics. In addition, the results show the intense Hg/protein ratio observed in the two proteins, which makes metalloproteins strong candidates for biomarkers of mercury exposure. [Figure not available: see fulltext.]. São Paulo State University (UNESP) Institute of Biosciences Federal University of Mato Grosso do Sul (UFMS) Biochemistry Department University of Nebraska University of São Paulo (USP) Local Development Dom Bosco Catholic University (UCDB) São Paulo State University (UNESP) Institute of Biosciences

Made available in DSpace on 2019-10-06T17:11:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01 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. School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) Institute of Biosciences São Paulo State University (UNESP) Federal University of Mato Grosso do Sul (UFMS) Biochemistry Department São Paulo University (USP) Biochemistry Department University of Nebraska-UNL School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) Institute of Biosciences São Paulo State University (UNESP)

Made available in DSpace on 2018-12-11T16:53:12Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-05-08 Predator fish can accumulate high levels of mercury, which qualifies them as potential indicators of this toxic metal. The predatory species Brachyplatystoma filamentosum, popularly known as filhote, is among the most consumed species in the Brazilian Amazon. Continuing the metalloproteomic studies of mercury in Amazonian fishes that have been developed in the last 5 years, the present paper provides the data of protein characterization associated with mercury in muscle and liver samples of filhote (Brachyplatystoma filamentosum) collected in the Madeira River, Brazilian Amazon. The mercury concentration in the muscle and liver samples was determined by graphite furnace atomic absorption spectrometry (GFAAS). The protein fraction was extracted in an aqueous medium, and later, a fractional precipitation procedure was performed to obtain the protein pellets. Then, the proteome of the tissue samples of this fish species was separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and a mercury mapping of the protein spots was carried out by GFAAS after acid digestion. Protein spots that had mercury were characterized by mass spectrometry with electrospray ionization in sequence (ESI-MS/MS) after tryptic digestion. It was possible to characterize 11 mercury-associated protein spots that presented biomarker characteristics and could be used to monitor mercury in fish species of the Amazon region. Thus, the metalloproteomic strategies used in the present study allowed us to characterize 11 mercury-associated protein spots. It should be noted that the protein spots identified as GFRP, TMEM186, TMEM57B, and BHMT, which have coordination sites for elements with characteristics of soft acids, such as mercury, can be used as biomarkers of mercury contamination in monitoring studies of this toxic metal in fish species from the Amazon region. School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) Institute of Biosciences São Paulo State University (UNESP) Biochemistry Department University of Nebraska School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) Institute of Biosciences São Paulo State University (UNESP)

Made available in DSpace on 2018-12-11T16:50:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-08-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) This paper presents a slurry sampling method for total mercury determination by graphite furnace atomic absorption spectrometry (GFAAS) in tissue of fish from the Amazon. The tissue samples were lyophilized and macerated, and then the slurry samples were prepared by putting 20 mg of tissue, added to a solution containing Triton X-100, Suprapur HNO3, and zirconium nitrate directly in sampling vials of a spectrometer. Mercury standard solutions were prepared under the same conditions as the slurry samples. The slurry samples and the mercury standard solutions were sonicated for 20 s. Twenty microliters of slurry samples were injected into the graphite tube, which contained an internal wall lined with tungsten carbide. Under these conditions, it was possible to thermally stabilize the mercury up to an atomization temperature of 1700 °C. The method was validated by mercury determination in reference materials DORM-4 and DOLT-4. The LOD and LOQ were 0.014 and 0.045 mg kg−1, respectively, and recovery percentages in relation to the concentration values were certified in the order of 98%. School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) Institute of Biosciences São Paulo State University (UNESP) Biochemistry Department University of Nebraska School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) Institute of Biosciences São Paulo State University (UNESP) FAPESP: 2010/51332-5 FAPESP: 2013/21297-1

Made available in DSpace on 2022-04-28T19:42:13Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-09-30 This study describes the association between meat tenderness and abundance of soluble muscle proteins in Nellore bulls (Bos indicus) using a proteomic approach. We evaluated shear force (SF) of Longissimus thoracis muscle 24 h after slaughter and selected three experimental groups of animals with moderately tender (TE; SF = 3.9 ± 0.7 kg), moderately tough (TO; SF = 5.6 ± 0.7 kg) and very tough meat (TO+; SF = 7.9 ± 1.4 kg). Proteome was investigated by two-dimensional electrophoresis (2D-PAGE) in combination with electrospray ionization-tandem mass spectrometry (ESI–MS/MS). The metabolic proteins triosephosphate isomerase (TPI1) and phosphoglucomutase 1 (PGM1), the structural protein profilin 1 (PFN1), and cytosol aminopeptidase (LAP3) were up-regulated (P < 0.05) in the TE meat group when compared to the TO and TO+ groups. Actin structural proteins (ACTA1, ACTB, and ACTG1), the oxidative stress protein peroxiredoxin (PRDX6, PRDX2, PRDX1, and PARK7), heat shock protein isoforms, and co-chaperones (CDC37 and STIP1) were up-regulated (P < 0.05) in the TO and TO+ meat groups. In addition, we also identified proteins PFN1, LAP3, PRDX1, PRDX2, HSPD1, and ARHGDIA to be associated with beef tenderness. The results reported herein demonstrated that meat tenderness in Nellore cattle depends on the modulation and expression of a set of proteins involved in different biological pathways. Significance: The manuscript entitled “Application of proteomic to investigate the different degrees of meat tenderness in Nellore breed” describes a classical proteomics work using two-dimensional gel electrophoresis (2D-PAGE), followed by mass spectrometry coupled to electrospray ionization ion trap (ESI-MS/MS) in order to understand the biochemical engineering involved in the process of meat tenderness. We evaluated shear force (SF) of Longissimus thoracis muscle samples of Nellore cattle (n = 90) and select three experimental groups of animals with moderately tender (TE; SF = 3.9 ± 0.7), moderately tough (TO; SF = 5.6 ± 0.7) and very tough meat (TO+; SF = 7.9 ± 1.4). The proteomic approach allowed observing that meat tenderness is influenced by structural proteins (ACTA1, ACTG1, ACTB, MYL1 and PFN1), co-chaperones (CDC37 and STIP1), heat shock proteins (HSP90AA1, HSP90AB1, HSPD1, HSPA1L, HSPA1A and HSPB1), regulatory protein (ARHGDIA), metabolic proteins (TPI1 and PGM1) and oxidative stress proteins (PRDX1, PRDX2, PRDX6, PARK7). Our results suggest that meat tenderness in Nellore depends on the modulation and expression of a set of proteins involved in different biological pathways. São Paulo State University (UNESP) College of Agriculture and Veterinary Science (FCAV), Jaboticabal Universidad de Ciencias Aplicadas y Ambientales (U.D.C.A) Redox Biology Center Department of Biochemistry University of Nebraska Institute of Bioscience (IB) São Paulo State University (UNESP), Botucatu São Paulo State University (UNESP) School of Veterinary Medicine, Araçatuba São Paulo State University (UNESP) School of Veterinary Medicine and Animal Science (FMVZ) Botucatu São Paulo State University (UNESP) College of Agriculture and Veterinary Science (FCAV), Jaboticabal Institute of Bioscience (IB) São Paulo State University (UNESP), Botucatu São Paulo State University (UNESP) School of Veterinary Medicine, Araçatuba São Paulo State University (UNESP) School of Veterinary Medicine and Animal Science (FMVZ) Botucatu

Made available in DSpace on 2018-12-11T16:47:41Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-12-01 Bioaccumulative metals such as mercury are found in increasing amounts in fish and their consumers. In the region of the Madeira River, in the Brazilian Amazon, mercury (Hg) is a predominant contaminant in the aquatic ecosystem. There is therefore a need to find specific biomarkers of mercury toxicity in fish to monitor contaminations. Here, mercury-bound proteins were identified in the liver tissues of fishes Mylossoma duriventre and Brachyplatystoma rousseauxii. Mercury was quantified in liver tissue, pellets and protein spots by graphite furnace atomic absorption. Proteins were fractionated by two-dimensional polyacrylamide gel electrophoresis and identified by mass spectrometry with electrospray ionization. We identified nine proteins linked to mercury and that presented biomarker characteristics of mercury. Among the proteins identified, isoforms of parvalbumin, ubiquitin-40S ribosomal protein S27a, brain-specific angiogenesis inhibitor 1-associated protein 2-like protein 2 and betaine–homocysteine S-methyltransferase 1 are notable for having the molecular function of binding to metallic ions. Institute of Biosciences São Paulo State University (UNESP) University of Nebraska-Lincoln School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) University of São Paulo (USP) College of Planaltina University of Brasília (UnB) Institute of Biosciences São Paulo State University (UNESP) School of Veterinary Medicine and Animal Science São Paulo State University (UNESP)

Recent studies have demonstrated the association of mercury (Hg) with some fish proteins, milk, and hair from individuals exposed to the element in the Amazon. However, few studies involve identifying biomarkers of mercury exposure. Therefore, the present study aimed to identify potential biomarkers of Hg exposure in fish. For this, the muscular tissues of two species of fish (Prochilodus lineatus and Mylossoma duriventre) that feed the Amazonian human population were analyzed. Through the analyses obtained by graphite furnace atomic absorption spectrometry (GFAAS), it was possible to identify four protein SPOTS where mercury was present. These SPOTS, identified by mass spectrometry (ESI-MS/MS), included parvalbumin and ubiquitin-40S ribosomal protein S27a, and these being metalloproteins with biomarker characteristics. In addition, the results show the intense Hg/protein ratio observed in the two proteins, which makes metalloproteins strong candidates for biomarkers of mercury exposure. Graphical Abstract.

Made available in DSpace on 2020-12-12T01:56:24Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-12-01 Understanding the effect of pesticides on the survival of honeybee colonies is important because these pollinators are reportedly declining globally. In the present study, we examined the changes in the head proteome of nurse honeybees exposed to individual and combined pesticides (the fungicide pyraclostrobin and the insecticide fipronil) at field-relevant doses (850 and 2.5 ppb, respectively). The head proteomes of bees exposed to pesticides were compared with those of bees that were not exposed, and proteins with differences in expression were identified by mass spectrometry. The exposure of nurse bees to pesticides reduced the expression of four of the major royal jelly proteins (MRJP1, MRJP2, MRJP4, and MRJP5) and also several proteins associated with carbohydrate metabolism and energy synthesis, the antioxidant system, detoxification, biosynthesis, amino acid metabolism, transcription and translation, protein folding and binding, olfaction, and learning and memory. Overall, when pyraclostrobin and fipronil were combined, the changes in protein expression were exacerbated. Our results demonstrate that vital proteins and metabolic processes are impaired in nurse honeybees exposed to pesticides in doses close to those experienced by these insects in the field, increasing their susceptibility to stressors and affecting the nutrition and maintenance of both managed and natural colonies. Grupo de Estudos em Apicultura e Meliponicultura Sustentável de Mato Grosso do Sul - GEAMS Federal University of Mato Grosso do Sul (UFMS) Faculty of Veterinary Medicine and Animal Science (FAMEZ) São Paulo State University (UNESP) Institute of Biosciences Department of Chemistry and Biochemistry Federal University of Mato Grosso do Sul (UFMS) Institute of Chemistry (INQUI) University of São Paulo (USP) Bauru Dental School Núcleo de Ensino Ciência e Tecnologia em Apicultura Racional (NECTAR) São Paulo State University (UNESP) School of Veterinary Medicine and Animal Science Department of Animal Production São Paulo State University (UNESP) Institute of Biosciences Department of Chemistry and Biochemistry Núcleo de Ensino Ciência e Tecnologia em Apicultura Racional (NECTAR) São Paulo State University (UNESP) School of Veterinary Medicine and Animal Science Department of Animal Production

Made available in DSpace on 2019-10-06T15:49:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-12-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) High concentrations of mercury found in soils, sediments, fish, and humans of the Amazon region have gained prominence in scientific studies during the last decade. However, studies related to the elucidation of mercury toxicity mechanisms in ichthyofauna at the molecular and metallomic levels that seek to elucidate physiological and functional aspects, as well as the search for biomarkers of mercury exposure, are still sparse. In the search for these answers, the present study analyzed the hepatic tissue proteome of the Arapaima gigas (pirarucu) fish species collected in the Jirau hydroelectric power plant reservoir in the state of Rondônia state, Brazil, in order to identify mercury-related metal-binding proteins and to elucidate their physiological and functional aspects. The proteomic profile of the hepatic tissue of Arapaima gigas was obtained by two-dimensional electrophoresis (2D-PAGE) and the presence of mercury was mapped in the protein SPOTS by graphite furnace atomic absorption spectrometry(GFAAS). Mercury was detected in 18 protein SPOTS with concentrations ranging from 0.13 ± 0.003 to 131.00 ± 3 mg kg−1. The characterization of the protein SPOTS associated with mercury was performed by electrospray ionisation tandem mass spectrometry (ESI–MS/MS), and 10 proteins were identified. Bioinformatics analyses showed that most of the proteins found linked to mercury were involved in cellular component processes and biological processes. For the most part, protein sequences have cellular functions comprising catalytic, binding, sense of localization, and metabolic processes. São Paulo State University (UNESP) School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) Institute of Biosciences University of São Paulo (USP) Institute of Chemistry (Inqui) Federal University of Mato Grosso do Sul, Brazil (UFMS) University of Nebraska (UNL) University of Brasília (UNB) College of Planaltina São Paulo State University (UNESP) School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) Institute of Biosciences FAPESP: 2016/19404-2 FAPESP: 2017/09466-3 CNPq: 304768/2018-19 CNPq: 404485/2016-2

Made available in DSpace on 2019-10-06T15:56:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01 This study aimed to evaluate the quality of the royal jelly produced by Apis mellifera bees in the presence of different iron concentrations (ferrous sulfate heptahydrate—0, 25, 50, and 100 mg L−1). Two-dimensional electrophoresis was used for the fractionation of royal jelly proteins, and iron level was quantified using flame atomic absorption spectrometry technique. The proteins were identified using electrospray ionisation mass spectrometry. Analysis of variance followed by the Tukey test (P < 0.05) was utilised. Dietary supplementation with mineral Fe affected the protein content and number of proteins in the experimental period. Further, the diet containing the highest iron concentration showed a greater number of spots containing iron, as well as in the abdomen of the bees. The most protein containing Fe were classified as major royal jelly proteins. These results showed that Fe influenced the quality of royal jelly and can improve its nutritional value. UNESP - São Paulo State University School of Veterinary Medicine and Animal Science UNESP - São Paulo State University Institute of Biosciences of Botucatu UNESP - São Paulo State University School of Veterinary Medicine and Animal Science UNESP - São Paulo State University Institute of Biosciences of Botucatu

O desenvolvimento urbano de Botucatu (SP) apresenta sólidas bases econômicas, ambientais e de saneamento, que contribuem para melhorar a qualidade de vida da população. Com a crise de 1929 e o declínio do café, Botucatu não se sustentou financeiramente para manter o tratamento de esgoto, e o Ribeirão Lavapés começou a receber o esgoto da cidade, que, somente em 2006, foi conduzido à estação de tratamento. Os objetivos deste estudo foram avaliar os parâmetros de qualidade de água do Lavapés antes e depois da construção da estação de tratamento de esgoto, e compará-los àqueles observados na foz do Rio Capivara, essencialmente rural. Abordam-se também aspectos de desenvolvimento do município quanto ao saneamento. A metodologia utilizada envolveu a revisão da literatura pertinente a qualidade de água antes e depois do tratamento, com níveis de referência baseados na Classe II (CONAMA). Com esses dados, obteve-se o Índice de Qualidade de Água (IQA) estabelecido pela National Sanitation Foundation e adaptado pela Cetesb. A qualidade das águas do Lavapés e de seus tributários melhorou desde a implantação do sistema de tratamento, apresentando IQA “Boa”, próximo de atingir a capacidade de resiliência. O Rio Capivara apresenta IQA “Ótima”, apesar da influência das áreas agricultáveis. A continuidade das ações de recuperação do Lavapés e da aplicação efetiva das políticas públicas nos vários setores do município reflete positivamente no Índice de Desenvolvimento Humano no Município (IDHM), hoje com avaliação “Muito Alto”. Além disso, os trabalhos de conscientização da população e a proteção dos recursos hídricos, aliados à reintegração dos focos pontuais de poluição ao sistema-tronco de tratamento e à recomposição das matas ripárias, serão vitais para as águas do Lavapés serem utilizadas no abastecimento público mesmo com a dinâmica crescente do município.

Made available in DSpace on 2020-12-12T01:08:47Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-04-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fish is an important source of protein, vitamins, and minerals. However, this food is also a major source of human exposure to toxic contaminants such as mercury. Thus, this paper aimed to evaluate mercury-binding proteins for possible application as biomarkers of mercury contamination in hepatic and renal tissues of Plagioscion squamosissimus (carnivorous fish) and Colossoma macropomum (omnivorous fish) from the Amazon region using metalloproteomic approach. The proteome of hepatic and renal tissues of fish species was separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and the mercury concentrations in protein spots were determined by graphite furnace atomic absorption spectrometry (GFAAS). Finally, the protein spots associated to mercury were characterized by electrospray ionization mass spectrometry (ESI-MS/MS). The activity of antioxidant enzymes (SOD, CAT, GPx, and GST) and lipid peroxidation (LPO) were also determined. The results showed that the highest concentrations of mercury were found in the carnivorous species (P. squamosissimus) and that the accumulation pattern of this metal was higher in hepatic tissues than in renal tissues for both species. A tendency was observed for greater enzymatic activity in the hepatic and renal tissues of P. squamosissimus, the species with the highest concentration of mercury. Only GPx activity in the kidney and GST in the liver were lower for the P. squamosissimus species, and this finding can be explained by the interaction of mercury with these enzymes. The data obtained by ESI-MS/MS allowed for the characterization of the protein spots associated to mercury, revealing proteins involved in energy metabolism, biomolecules transport, protein synthesis and degradation, cell differentiation, gene regulation, and the antioxidant system. The results obtained in the present study can contribute to understanding the physiological processes underlying mercury toxicity and have provided new perspectives on possible candidates for mercury contamination biomarkers in fish. São Paulo State University (UNESP) School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) Institute of Biosciences University of Nebraska (UNL) University of Brasília (UNB) College of Planaltina University of São Paulo (USP) Institute of Chemistry (INQUI) Federal University of Mato Grosso do Sul Campo Grande (UFMS) São Paulo State University (UNESP) School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) Institute of Biosciences CNPq: 303719/2014-1 CNPq: 30478/2018-9 CNPq: 404485/2016-2

Made available in DSpace on 2020-12-10T19:43:00Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-03-20 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Sustainable Energy P&D project of Brazil S.A. 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. Sao Paulo State Univ Julio de Mesquita Filho, Dept Chem & Biochem, Botucatu, SP, Brazil Univ Sao Paulo, Bauru Fac Dent, Sao Paulo, SP, Brazil Univ Brasilia, Brasilia, DF, Brazil Sao Paulo State Univ Julio de Mesquita Filho, Dept Chem & Biochem, Botucatu, SP, Brazil FAPESP: 2014/02638-5 FAPESP: 2013/21297-1 FAPESP: 2016/19404-2 CNPq: 304768/2018-9 CNPq: 404485/2016-2 Sustainable Energy P&D project of Brazil S.A.: 6631-0001/2012 Sustainable Energy P&D project of Brazil S.A.: 004/13

Made available in DSpace on 2018-12-11T16:55:15Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-01-01 Proteomics studies can be used to identify proteins that affect feed efficiency traits, related to cost and profitability of meat production. We used a proteomic approach based on two-dimensional electrophoresis (2D-PAGE) in combination with mass spectrometry (ESI-MS) to study liver samples of Nellore bulls divergently ranked according to residual feed intake (RFI). The study showed that 71 protein spots were expressed differentially (P < 0.05) among RFI groups and 47 were identified by ESI-MS. In RFI, efficient animals (low RFI) eat less than predictions, based on their weights and growth rate, while inefficient animals (high RFI) that eat more than predicted. Data from 18 animals (9 high vs. 9 low RFI) aged 24–26 months in feedlot finishing were used. Immediately after slaughter, liver samples were collected and protein extracts were separated. The gels of RFI groups were scanned and the images analyzed, whereby we found 279 and 215 liver protein spots in high and low RFI bulls, respectively. The proteins identified were related to the following biological functions: (I) oxygen transport and blood flow; (II) mitochondrial function and energy metabolism; (III) amino acid metabolism, ion transport, and cell survival. The study suggests hemoglobin subunit beta and heat shock protein 71 kDa and as molecular markers to study FE in Nellore cattle. Moreover, proteins such as 3-ketoacyl-CoA thiolase and glutamate dehydrogenase 1 were found in liver from high and low RFI animals, respectively. Such protein expression could be associated with changes in the oxidative capacity of RFI phenotypes. Department of Animal Science Luiz de Queiroz College of Agriculture University of São Paulo (ESALQ-USP) Animal Science and Pastures Institute - Instituto de Zootecnia (IZ) Centro APTA Bovinos de Corte Institute of Biosciences São Paulo State University (UNESP) Institute of Biosciences São Paulo State University (UNESP)

Made available in DSpace on 2018-12-11T16:47:27Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-02-01 Mercury is a potentially toxic element that is present in the environment of the Brazilian Amazon and is responsible for adverse health effects in humans. This study sought to assess possible protein biomarkers of mercury exposure in breast milk samples from lactating women in the Madeira and Negro Rivers in the Brazilian Amazon. The mercury content of hair samples of lactating women was determined, and the proteome of breast milk samples was obtained using two-dimensional electrophoresis after protein precipitation with acetone. Mercury measurements of protein spots obtained via protein fractionation were performed by graphite furnace atomic absorption spectrometry (GFAAS), and it was observed that mercury is linked to proteins with molecular masses in the range of 14–26 kDa. The total mercury concentration was also determined by GFAAS in unprocessed milk, lyophilized milk, and protein pellets, with the purpose of determining the mercury mass balance in relation to the concentration of this element in milk and pellets. Approximately 85 to 97% of mercury present in the lyophilized milk from samples of lactating women of the Madeira River is bound in the protein fraction. From lactating women of the Negro River, approximately 49% of the total mercury is bound in the protein fraction, and a difference of 51% is bound in the lipid fraction. Pontifical Catholic University of Goiás (PUC) Institute of Biosciences São Paulo State University (UNESP) São Paulo State University (UNESP) College of Planaltina University of Brasília (UnB) Institute of Biosciences São Paulo State University (UNESP) São Paulo State University (UNESP)

This study used metalloproteomic techniques to characterize mercury (Hg)-bound proteins in the muscle and liver tissue of Tucunare (Cichla spp.) collected at the Jirau Hydroelectric Power Plant in Madeira River Basin, Brazil. The proteome of the muscle and liver tissue was obtained after two steps of fractional precipitation and separating the proteins by 2-D polyacrylamide gel electrophoresis. Hg was identified and quantified in the protein spots by graphite furnace atomic absorption spectrometry after acid mineralization in an ultrasound bath. Hg with a molecular weight

Made available in DSpace on 2018-12-11T16:48:54Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-05-01 This study presents data on the extraction and characterization of proteins associated with mercury in the muscle and liver tissues of jaraqui (Semaprochilodus spp.) from the Madeira River in the Brazilian Amazon. Protein fractionation was carried out by two-dimensional electrophoresis (2D-PAGE). Mercury determination in tissues, pellets, and protein spots was performed by graphite furnace atomic absorption spectrometry (GFAAS). Proteins in the spots that showed mercury were characterized by electrospray ionization tandem mass spectrometry (ESI-MS/MS). The highest mercury concentrations were found in liver tissues and pellets (426 ± 6 and 277 ± 4 μg kg−1), followed by muscle tissues and pellets (132 ± 4 and 86 ± 1 μg kg−1, respectively). Mercury quantification in the protein spots allowed us to propose stoichiometric ratios in the range of 1–4 mercury atoms per molecule of protein in the protein spots. The proteins characterized in the analysis by ESI-MS/MS were keratin, type II cytoskeletal 8, parvalbumin beta, parvalbumin-2, ubiquitin-40S ribosomal S27a, 39S ribosomal protein L36 mitochondrial, hemoglobin subunit beta, and hemoglobin subunit beta-A/B. The results suggest that proteins such as ubiquitin-40S ribosomal protein S27a, which have specific domains, possibly zinc finger, can be used as biomarkers of mercury, whereas mercury and zinc present characteristics of soft acids. Institute of Bioscience of Botucatu São Paulo State University (UNESP), Rubião Júnior Pontifical Catholic University of Goiás (PUC) Bauru Dental School University of São Paulo-USP Institute of Bioscience of Botucatu São Paulo State University (UNESP), Rubião Júnior

Made available in DSpace on 2018-11-26T17:16:36Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-03-01 Proteins play crucial roles in biological systems, thus studies comparing the protein pattern present in a healthy sample with an affected sample have been widely used for disease biomarker discovery. Although proteins containing metal ions constitute only a small proportion of the proteome, they are essential in a multitude of structural and functional processes. The correct association between metal ions and proteins is essential because this binding can significantly interfere with normal protein function. Employment of a metalloproteomic study of liver samples from diabetic rats permitted determination of the differential abundance of copper-, selenium-, zinc- and magnesium-associated proteins between diabetic, diabetic treatment with insulin and non-diabetic rats. Proteins were detected by ESI-MS/MS. Seventy-five different proteins were found with alterations in the metal ions of interest. The most prominent pathways affected under the diabetic model included: amino-acid metabolism and its derivates, glycogen storage, metabolism of carbohydrates, redox systems and glucose metabolism. Overall, the current methods employed yielded a greater understanding of metal binding and how type 1 diabetes and insulin treatment can modify some metal bonds in proteins, and therefore affect their mechanism of action and function. (C) 2017 Elsevier B.V. All rights reserved. Sao Paulo State Univ, Inst Biosci Botucatu, Dept Chem & Biochem, Botucatu, SP, Brazil Univ Nebraska, Dept Biochem, Lincoln, NE 68583 USA Univ Sao Paulo, Bauru Dent Sch, Bauru, SP, Brazil Sao Paulo State Univ, Inst Biosci Botucatu, Dept Chem & Biochem, Botucatu, SP, Brazil

Made available in DSpace on 2018-12-11T17:12:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-11-01 Type 1 diabetes is characterized by hyperglycemia, which in the chronic stage is associated with abnormalities in lipids, protein and, carbohydrate metabolism, as well as oxidative stress. New strategies for prevention and treatment are needed, as type 1 diabetes affects life quality and survival, and involves high-cost treatment. Proteomic and metalloproteomic studies can elucidate the functional and physiological aspects of biomolecules. In the present study, differential proteomics was used to identify potential biomarkers of diabetes in rat plasma associated with copper, selenium, zinc, and magnesium fractionation in control and diabetic rats, as well as diabetic rats treated with insulin. 2D-PAGE was used in the plasma protein fractionation; graphite furnace atomic absorption spectrometry (GFAAS) and flame atomic absorption spectrometry (FAAS) were used for quantitative determination of copper, magnesium, selenium, and zinc in the spots that showed different expression; and protein spots were characterized by electrospray ionization-tandem mass spectrometry (ESI–MS/MS) after tryptic digestion. ESI–MS/MS analysis characterized 35 different proteins, indicating alpha-1-macroglobulin and haptoglobulin as potential candidates as biomarkers for diabetes treated with insulin; also, 2′-deoxynucleoside 5′-phosphate N-hydrolase 1, transmembrane protein 11, serum amyloid P component, vitamin D-binding protein, and biliverdin reductase were identified as potential candidates as biomarkers for uncontrolled diabetes. Department of Chemistry and Biochemistry Institute of Bioscience São Paulo State University (UNESP) Bauru Dental School University of São Paulo – USP, Bauru Department of Chemistry and Biochemistry Institute of Bioscience São Paulo State University (UNESP)

Three feeding frequencies (24, 48 and 96 meals/day) and three feeding periods (daytime, nighttime and daytime/nighttime) were evaluated in this experiment. Bullfrogs ( Lithobates catesbeianus ), with average weight of 78.6 ± 15.7 g, were distributed in 36 pens at a density of 88 frogs/m 2 . The pens, made of polypropylene and screened floor, were installed inside cages, linearly distributed in a 2000 m 2 pond with 5% water renewal. The frogs were fed daily by automatic feeders with extruded commercial feed for carnivore fish. The experiment lasted 90 days, corresponding to the autumn and winter seasons. Feeding frequency and period interacted on the frogs’ average weight (AW) values. At the end of the experiment, in the daytime and daytime/nighttime periods, there was no difference of the AW values between the evaluated feeding frequencies; however, in the nighttime period, the frogs’ AW was greater for the frequency of 48 meals/day. Separately, the feeding frequency and period influenced the result of apparent feed conversion ratio (FCR) which was better with the frequency of 96 meals/day with average value of 2.13, and average value of 2.23 at nighttime. There was no difference between the treatments regarding the coefficient of apparent digestibility of dry matter, crude protein and crude energy. The coefficients of apparent availability of calcium and phosphorus were higher with the feeding frequency of 48 meals/day, regardless of the feeding period. The results showed that bullfrogs can be fed at daytime as well as at nighttime and that the highest feeding frequencies improved FCR in the production of this species. For bullfrog rearing in cages, it is recommended feeding frequency of 96 meals/day at daytime and daytime/nighttime periods and 48 meals/day at nighttime.

Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:29:47Z No. of bitstreams: 0 Made available in DSpace on 2014-05-27T11:29:47Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-06-27 In the present study, a simple, rapid and sensitive method was developed for the determination of mercury concentrations in the muscle tissue of fish from the Brazilian Amazon using graphite furnace atomic absorption spectrometry (GFAAS) following acid mineralization of the samples in an ultrasonic cold water bath. Using copper nitrate as a chemical modifier in solution and sodium tungstate as permanent modifier, we were able to attain thermal stabilization of the mercury up to the atomisation temperature of 1600 °C in the GFAAS assay. The calculated limits of detection (LOD) and quantification (LOQ) were 0.014 and 0.047 mg kg-1, respectively. © 2013 Elsevier Ltd. All rights reserved. UNESP - São Paulo State University Biosciences Institute, Botucatu, SP UNESP - São Paulo State University Faculty of Veterinary Medicine and Animal Science, Botucatu, SP University of Alagoas - IQB/UFAL, Maceió, Al University of Brasília Faculdade de Planaltina Distrito Federal National Institute of Bioanalytical Sciences and Technology UNICAMP - Campinas State University, Campinas, SP UNESP - São Paulo State University Biosciences Institute, Botucatu, SP UNESP - São Paulo State University Faculty of Veterinary Medicine and Animal Science, Botucatu, SP

This paper presents preliminary findings for a metallomics study of mercury in the muscle of the fish species from Amazonas, Brazil, after protein separation by two-dimensional polyacrylamide gel electrophoresis and subsequent evaluation of mercury by synchrotron radiation X-ray fluorescence. The fluorescence spectra revealed mercury in two protein spots. The mercury-containing protein spots showed molecular weights of 20.8 ± 0.7 and 19.8 ± 0.5 kDa and isoelectric points of 5.6 ± 0.2 and 7.5 ± 0.3, respectively.

Made available in DSpace on 2018-12-11T16:58:59Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-12-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) The feeds supplied to farmed fish should be rich in protein and minerals, such as animal protein, but more expensive alternatives comprising vegetable protein have emerged in recent years. However, the minerals essential to the proper development of fish must be assessed. To determine whether the minerals supplied in the diet are being fully exploited, the equation of the apparent digestibility coefficient is typically used with an external marker such as chromium. Moreover, the concentrations of the metals of interest are usually obtained using an acid digestion in the digester block; however, this digestion is harmful to both the analyst and the environment. An alternative to this digestion is described in this work. Four types of feeds were made with different types of protein substitutes. Department of Chemistry and Biochemistry Institute of Bioscience São Paulo State University (UNESP), Rubião Junior District College of Veterinary and Animal Science São Paulo State University Department of Chemistry and Biochemistry Institute of Bioscience São Paulo State University (UNESP), Rubião Junior District College of Veterinary and Animal Science São Paulo State University FAPESP: 2008/51377-9

This study aimed to identify metalloproteins that lose their metal ions in the presence of mercury (Hg) and bind to Hg in breast milk samples collected from the riverine population of the Madeira River, a tributary of the Amazon River. Initially, total Hg was determined from the hair of lactating women to identify individuals who were contaminated followed by a proteomic analysis of breast milk samples through two-dimensional polyacrylamide gel electrophoresis after acetone precipitation. The presence of Hg in the obtained protein spots was determined by cold vapor atomic absorption spectrometry and cold vapor atomic fluorescence spectrometry. These determinations indicated the presence of Hg in one protein spot, which was then characterized through electrospray ionization tandem mass spectrometry. Based on searches in the UniProt database, this protein spot was identified as lysozyme C.