Marcos Hikari Toyama | Universidade Estadual Paulista "Júlio de Mesquita Filho" (original) (raw)

Papers by Marcos Hikari Toyama

PLOS ONE

Multiple drug resistance (MDR) bacterial strains are responsible by 1.2 million of human deaths a... more Multiple drug resistance (MDR) bacterial strains are responsible by 1.2 million of human deaths all over the world. The pathogens possess efficient enzymes which are able to mitigate the toxicity of reactive oxygen species (ROS) produced by some antibiotics and the host immune cells. Among them, the bacterial peroxiredoxin alkyl hydroperoxide reductase C (AhpC) is able to decompose efficiently several kinds of hydroperoxides. To decompose their substrates AhpC use a reactive cysteine residue (peroxidatic cysteine—CysP) that together with two other polar residues (Thr/Ser and Arg) comprise the catalytic triad of these enzymes and are involved in the substrate targeting/stabilization to allow a bimolecular nucleophilic substitution (SN2) reaction. Additionally to the high efficiency the AhpC is very abundant in the cells and present virulent properties in some bacterial species. Despite the importance of AhpC in bacteria, few studies aimed at using natural compounds as inhibitors of t...

INDICE viii CAPÍTULO 3. …………………………………………………………………………… 69 Crystallography and bioinformatics in th... more INDICE viii CAPÍTULO 3. …………………………………………………………………………… 69 Crystallography and bioinformatics in the study of platelet aggregation and the enzymatic and myotoxic activities of BjVIII: a new PLA2 Bthtx-I-like isoform. …………………………………...… 69 CAPÍTULO 4. …………………………………………………………………………… 99 Evaluation of two synthetic coumarins on the structure and Biological activity of myotoxic Lys49 sPLA2 from Bothrops jararacussu 99 CONSIDERAÇÕES FINAIS. ………………………………………………..………… REFERÊNCIAS BIBLIOGRÁFICAS. ………………………………………………… ANEXOS. ………………………………………………………………………………...

Antioxidants, 2021

Typical 2-Cys peroxiredoxins (2-Cys Prx) are ubiquitous Cys-based peroxidases, which are stable a... more Typical 2-Cys peroxiredoxins (2-Cys Prx) are ubiquitous Cys-based peroxidases, which are stable as decamers in the reduced state, and may dissociate into dimers upon disulfide bond formation. A peroxidatic Cys (CP) takes part of a catalytic triad, together with a Thr/Ser and an Arg. Previously, we described that the presence of Ser (instead of Thr) in the active site stabilizes yeast 2-Cys Prx as decamers. Here, we compared the hyperoxidation susceptibilities of yeast 2-Cys Prx. Notably, 2-Cys Prx containing Ser (named here Ser-Prx) were more resistant to hyperoxidation than enzymes containing Thr (Thr-Prx). In silico analysis revealed that Thr-Prx are more frequent in all domains of life, while Ser-Prx are more abundant in bacteria. As yeast 2-Cys Prx, bacterial Ser-Prx are more stable as decamers than Thr-Prx. However, bacterial Ser-Prx were only slightly more resistant to hyperoxidation than Thr-Prx. Furthermore, in all cases, organic hydroperoxide inhibited more the peroxidase a...

Bothrops is a genus of pit vipers (snakes) endemic to Central and South America. Snake envenomati... more Bothrops is a genus of pit vipers (snakes) endemic to Central and South America. Snake envenomation (Bothrops genus) specifically Bothropoides pauloensis, a venomous snake popularly known as “Jararaca pintada”, which is found in the Brazilian territory or more commonly in the southwest of the state of Sao Paulois. Acute kidney injury is one of the complications observed in Bothrops snakebite with relevant morbidity and mortality. Here, we showed the cytotoxic effect of Bothropoides pauloensis venom on MDCK cells, intracellular mitochondrial membrane potentiality (ΔΨm), BpV regulated Reactive Oxygen Species (ROS) and activation of caspases 3 and 7 in MDCK cells. MDCK cells were treated with different concentrations of B. pauloensis venom for 24 h and cell death were measured with annexin V and PI staining and detected by flow cytometry. MDCK cells were treated with two different concentrations (IC50 and 2 × IC50) of BpV after 24 h. All data are expressed as mean ± SEM of three indepe...

Free Radical Biology and Medicine, 2020

Virtual population analysis requires information on the fishing mortality rate (or abundance) for... more Virtual population analysis requires information on the fishing mortality rate (or abundance) for one age group from each cohort. In many cases available data are insufficient to estimate these rates for every age group and structural assumptions must be imposed to reduce the number of effective parameters. Past assessments of western Atlantic bluefin tuna (Thunnus thynnus) have reduced the number of parameters by assuming pre-specified values for the ratios of the fishing mortality rates on adjacent age groups. More recent bluefin tuna assessments have estimated terminal fishing mortality rates (F) on each age with a constraint that restricts change from one year to the next. We explore the implications of these methods of estimating terminal-year F through retrospective analyses of the 2006 bluefin tuna assessment and through stochastic simulations. The use of pre-specified ratios for F created strong retrospective biases and may have led to overly optimistic projections. Constraining annual changes in the terminalyear F appeared to mute retrospective patterns and resulted in abundance projections less prone to spurious initial leaps. Simulation results indicate that the constraint improves estimation, particularly with moderate to low interannual changes in selectivity.

Frontiers in Cellular and Infection Microbiology, 2020

Journal of the Brazilian Chemical Society, 2020

The pyrolysis of waste tire has been proposed as an alternative route to its recycling producing ... more The pyrolysis of waste tire has been proposed as an alternative route to its recycling producing residual oils that can be used as fuels. In this work, a physicochemical process to obtain carbonbased dots (CBD) from carbon black (CB L) derivatives of tires is described. For this, the CB L was treated with different acids to produce CB L1 and CB L2 particles suspension. The pH of the suspension was adjusted to 5.6 to precipitate remaining impurities. Afterwards, the supernatant was ultra-centrifuged and the yellow pale solid, the purified CBD obtained by roto evaporation, was characterized by ultraviolet-visible spectrophotometer (UV-Vis), photoluminescence (PL) at 320-380 nm, high-resolution transmission electron microscopy (HRTEM), scanning electron microscopy (SEM), thermogravimetric analysis (TGA), energy-dispersive X-ray spectroscopy (EDX), Fourier transform infrared microscopy (µFTIR), and X-ray photoelectron spectroscopy (XPS). This material showed the typical excitation wavelength dependent emission and a PL quantum yield of 19.29% and was constituted by a well-defined spherical shaped and single sheets of "graphene quantum dots" with undefined shape, with potential applications in optoelectronic and fluorescence imaging devices.

Biotechnology Letters, 2020

Acute lymphoblastic leukaemia (ALL) affects lymphoblastic cells and is the most common neoplasm d... more Acute lymphoblastic leukaemia (ALL) affects lymphoblastic cells and is the most common neoplasm during childhood. Among the pharmaceuticals used in the treatment protocols for ALL, Asparaginase (ASNase) from Escherichia coli (EcAII) is an essential biodrug. Meanwhile, the use of EcAII in neoplastic treatments causes several side effects, such as immunological reactions, hepatotoxicity, neurotoxicity, depression, and coagulation abnormalities. Commercial EcAII is expressed as a recombinant protein, similar to novel enzymes from different organisms; in fact, EcAII is a tetrameric enzyme with high molecular weight (140 kDa), and its Keywords Escherichia coli ASNaseII (EcAII) Á Acute lymphoblastic leukemia (ALL) Á Recombinant enzyme Á Low temperature expression Á Functional and structural evaluation

Natural Product Communications, 2011

Cytotoxicity assays are needed for the screening of natural products with potential anti-inflamma... more Cytotoxicity assays are needed for the screening of natural products with potential anti-inflammatory. The purpose of this study was to compare the basal cytotoxicity of active ingredients extracted from plants of the Brazilian “cerrado”. The viability was assayed with the neutral red uptake assay in Mac Coy cells after 24h of exposition. The dose evaluated was 50 μg/μL. The test substances were: cinnamic acid, p-coumaric acid, chlorogenic acid, syringic acid, vannilic acid, homogentisic acid, scandenin, palustric acid, diosgenin, cabraleone. Studies of cytotoxicity demonstrated that all active compounds evaluated have low toxicity in vitro. The substances showed cell viability above 60% for the concentration used. However, the cinnamic acid, sacandenin and palustric acid showed highest toxicity with a 50% reduction in cell viability for the dose of 50 μg/μL. Cytotoxic screening results are useful to estimate the best concentrations of those compounds with potential anti-inflammator...

The Journal of Nutrition, 2003

Undernutrition has been shown to affect the autonomic nervous system, leading to permanent altera... more Undernutrition has been shown to affect the autonomic nervous system, leading to permanent alterations in insulin secretion. To understand these interactions better, we investigated the effects of carbamylcholine (CCh) and phorbol 12-myristate 13-acetate (PMA) on insulin secretion in pancreatic islets from rats fed a normal (17%; NP) or low (6%; LP) protein diet for 8 wk. Isolated islets were incubated for 1 h in Krebs-bicarbonate solution containing 8.3 mmol glucose/L, with or without PMA (400 nmol/L) and CCh. Increasing concentrations of CCh (0.1-1000 mol/L) dose dependently increased insulin secretion by islets from both groups of rats. However, insulin secretion by islets from rats fed the NP diet was significantly higher than that of rats fed the LP diet, and the dose-response curve to CCh was shifted to the right in islets from rats fed LP with a 50% effective concentration (EC 50) of 2.15 Ϯ 0.7 and 4.64 Ϯ 0.1 mol CCh/L in islets of rats fed NP and LP diets, respectively (P Ͻ 0.05). PMA-induced insulin secretion was higher in islets of rats fed NP compared with those fed LP. Western blotting revealed that the protein kinase (PK)C␣ and phospholipase (PL)C␤ 1 contents of islets of rats fed LP were 30% lower than those of islets of rats fed NP (P Ͻ 0.05). In addition, PKC␣ mRNA expression was reduced by 50% in islets from rats fed LP. In conclusion, a reduced expression of PKC␣ and PLC␤ 1 may be involved in the decreased insulin secretion by islets from LP rats after stimulation with CCh and PMA.

Natural Products and Bioprospecting, 2018

We recognize the chemical composition of the acetonic extract of Rhizophora mangle barks (AERM) u... more We recognize the chemical composition of the acetonic extract of Rhizophora mangle barks (AERM) using mass spectrometry analysis [liquid chromatography (LC)-ESI-IT-MS/MS and matrix-assisted laser desorption/ionization-time of flight-MS (MALDI-TOF)]. Analgesic activity was evaluated by formalin and tail-flick experimental assays. Anti-inflammatory activity was performed by paw edema test induced by carrageenan and 48/80 compounds. The first series of experiments involved [LC]-FIA-IT-MS/MS with 11 separated catechins derivatives until degree of polymerization 3 (DP3). The spectra obtained by MALDI-TOF analysis of the AERM presented two homologous series: one based on polymers of m/z 288 Da increments (up to DP12) and another series based on polymers of m/z [288 ? 162] Da increments (up to DP11). In addition to these series of flavan-3-ol, each DP had a subset of masses with a variation of-16 Da (homologous series of afzelechins-m/z 873-3465 Da) and ? 16 Da (homologous series of gallocatechins-m/ z 905-3497 Da). A similar pattern with homologous series of gallocatechins and afzelechins could also be observed for a fifth and a sixth monohexoside series: glucogallocatechins (m/z 779-3371) and glucoafzelechins (m/z 747-3339). The intraperitoneal administration of different doses of AERM (50, 150 and 300 lg mL-1) have a morphine-like effect and intense anti-inflammatory activity.

Scientific Reports, 2016

Typical 2-Cys Peroxiredoxins (2-Cys Prxs) reduce hydroperoxides with extraordinary rates due to a... more Typical 2-Cys Peroxiredoxins (2-Cys Prxs) reduce hydroperoxides with extraordinary rates due to an active site composed of a catalytic triad, containing a peroxidatic cysteine (CP), an Arg, and a Thr (or Ser). 2-Cys Prx are involved in processes such as cancer; neurodegeneration and host-pathogen interactions. During catalysis, 2-Cys Prxs switch between decamers and dimers. Analysis of 2-Cys Prx structures in the fully folded (but not locally unfolded) form revealed a highly conserved, non-conventional hydrogen bond (CH-π) between the catalytic triad Thr of a dimer with an aromatic residue of an adjacent dimer. In contrast, structures of 2-Cys Prxs with a Ser in place of the Thr do not display this CH-π bond. Chromatographic and structural data indicate that the Thr (but not Ser) destabilizes the decamer structure in the oxidized state probably through steric hindrance. As a general trend, mutations in a yeast 2-Cys Prx (Tsa1) favoring the dimeric state also displayed a decreased ca...

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Nihon Ika Daigaku zasshi, 1965

African Journal of Pharmacy and Pharmacology, 2014

Pain is one of the most common reasons for patients to seek medical care. Bee Apis mellifera veno... more Pain is one of the most common reasons for patients to seek medical care. Bee Apis mellifera venom (AMV) has traditionally been used to treat inflammatory diseases and the alleviation of pain. Herein, we aimed to investigate the visceral antinociceptive potential of A. mellifera bee venom and its possible mechanism of action. Acetic acid-induced writhing assay was used in mice to determine the degree of visceral antinociception. Visceral antinociceptive activity was expressed as the reduction in the number of abdominal constrictions. Mice received an intraperitoneal injection of acetic acid after administration of AMV (0.08 or 0.8 mg/kg; intraperitoneally (i.p.)). In mechanistic studies, separate experiments were realized to examine the role of α2-receptors, nitric oxide, calcium channels, K + ATP channel activation, TRPV1 and opioid receptors on the visceral antinociceptive effect of AMV (0.8 mg/kg), using appropriate antagonists, yohimbine (2 mg/kg), L-NG-Nitroarginine methyl ester (L-NAME, 10 mg/kg), verapamil (5 mg/kg), glibenclamide (5 mg/kg), ruthenium red (3 mg/kg) or naloxone (2 mg/kg). AMV presented visceral antinociceptive activity in both doses tested (0.08 and 0.8 mg/Kg). Visceral antinociceptive effect of AMV was resistant to all the antagonists used. Mice showed no significant alterations in locomotion frequency, indicating that the observed antinociception is not a consequence of motor abnormality. Although AMV efficient diminished the acetic acid-evoked pain-related behavior, its mechanism is unclear from this study and future studies are needed to verify how the venom exerts its antinociceptive action.

American journal of physiology. Endocrinology and metabolism, 2003

Short-term muscle denervation is a reproducible model of tissue-specific insulin resistance. To i... more Short-term muscle denervation is a reproducible model of tissue-specific insulin resistance. To investigate the molecular basis of insulin resistance in denervated muscle, the downstream signaling molecules of the insulin-signaling pathway were examined in intact and denervated soleus muscle of rats. Short-term denervation induced a significant fall in glucose clearance rates (62% of control, P < 0.05) as detected by euglycemic hyperinsulinemic clamp and was associated with a significant decrease in insulin-stimulated tyrosine phosphorylation of the insulin receptor (IR; 73% of control, P < 0.05), IR substrate 1 (IRS1; 69% of control, P < 0.05), and IRS2 (82% of control, P < 0.05) and serine phosphorylation of Akt (39% of control, P < 0.05). Moreover, denervation reduced insulin-induced association between IRS1/IRS2 and p85/phosphatidylinositol (PI) 3-kinase. Nevertheless, denervation caused an increase in PI 3-kinase activity associated with IRS1 (275%, P < 0.05) ...

Toxicon, 1998

Bothtrops moojeni snake venom was fractionated on a CM-Sepharose column which was previously equi... more Bothtrops moojeni snake venom was fractionated on a CM-Sepharose column which was previously equilibrated with 0.05 M ammonium bicarbonate buffer at pH 8.0 and subsequently eluted with an ammonium bicarbonate concentration gradient from 0.05 to 0.5 M at constant pH (8.0) and temperature (25 degrees C). The fraction which eluted last (M-VI) showed, after direct lyophilization, a single band by polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE, indicating an approximate Mr of 14000 and 27000, in the presence and absence of dithiothreitol, respectively. Its amino acid composition revealed a high level of hydrophobic and basic amino acids as well as 14 half-cystine residues. Its isoelectric point and extinction coefficient (E(1.0 mg/ml) (1.0 cm) at 278 nm and pH 7.0) were 8.2 and 1.170, respectively. M-VI was devoid of phospholipase A2 (PLA2) activity on egg yolk, as well as of hemorrhagic, anticoagulant and coagulant activities, but could induce drastic necrosis on skeletal muscle fibres as well as rapid and transient edema on the rat paw. Its N-terminal sequence: SLFELGKMILQETGKNPAKSYGVYGCNCGVGGRGKPKDATDRCCYVHKCCYK... revealed high homology with other Lys 49 PLA2-like myotoxins from other bothropic venoms. Orthorhombic crystals of M-VI, which diffracted to a maximal resolution of 1.6 A, were obtained and indicated the presence of a dimer in the asymmetrical unit.

Journal of Venomous Animals and Toxins including Tropical Diseases, 2011

In the present study, the effects of Polybia paulista venom (PPV) on renal and vascular tissues w... more In the present study, the effects of Polybia paulista venom (PPV) on renal and vascular tissues were investigated. Isolated kidneys perfused with PPV (1 and 3 µg/mL) had increased perfusion pressure, renal vascular resistance , urinary flow, and glomerular filtration rate; and reduced sodium tubular transport. Histological evaluation demonstrated deposits of proteins in Bowman's space and tubular lumen, and focal areas of necrosis. The venom promoted a cytotoxic effect on Madin-Darby canine kidney (MDCK) cells. A significant increase in lactic dehydrogenase levels was observed in response to venom exposure. In isolated mesenteric vascular beds, pressure and vascular resistance augmented in a dose-dependent manner. PPV increased the contractility of aortic rings maintained under basal tension. This contractile response was inhibited when preparations were maintained in Ca 2+-free medium. Likewise, verapamil, a voltage-gated calcium channel blocker, also inhibited the contractile response. In this study, phentolamine, a blocker of α-adrenergic receptor blocker, significantly reduced the contractile effect of PPV in the aortic ring. In conclusion, PPV produced nephrotoxicity, which suggests a direct effect on necrotic cellular death in renal tubule cells. The vascular contractile effect of PPV appears to involve calcium influx through voltage-gated calcium channels via adrenergic regulation.

The Journal of Nutritional Biochemistry, 2008

Intrauterine growth restriction is associated with chronically elevated levels of serum fatty aci... more Intrauterine growth restriction is associated with chronically elevated levels of serum fatty acids and reduced glucose-stimulated insulin secretion. Lipid metabolism in pancreatic beta cells is critical for the regulation of insulin secretion, and the chronic exposure to fatty acids results in higher palmitate oxidation rates and an altered insulin response to glucose. Using a rat model of isocaloric protein restriction, we examined whether pre- and postnatal protein malnutrition influences the properties of pancreatic islet carnitine palmitoyltransferase-1 (liver isoform, L-CPT-1), a rate-limiting enzyme that regulates fatty acid oxidation in mitochondria. The activity of L-CPT-1 in pancreatic islets increased in the low protein (LP), although the L-CPT-1 mRNA levels were unaffected by malnutrition. The susceptibility of enzyme to inhibition by malonyl-CoA was unaltered and the content of malonyl-CoA was reduced in LP cells. Because the mitochondrial oxidation of fatty acids is related to the altered expression of a number of genes encoding proteins involved in insulin secretion, the levels of expression of insulin and GLUT-2 mRNA were assessed. A reduced expression of both genes was observed in malnourished rats. These results provide further evidence that increased L-CPT-1 activity and changes in gene expression in pancreatic islets may be involved in the reduced insulin secretion seen in malnourished rats.