RESUMO
Intravascular hemolysis (IH) contributes to the development of endothelial dysfunction (ED) in sickle cell anemia (SCA), and the effects of hydroxyurea (HU, the only approved drug that decreases the frequency and severity of vaso-oclussive crises) on IH and ED in SCA remain unclear. We evaluated and compared the markers of IH among steady-state adult Brazilians with SCA and HbAA individuals. Overall, this cross-sectional study enrolled 30 SCA patients not receiving HU therapy (HbSS), 25 SCA patients receiving HU therapy (HbSS_HU), and 32 HbAA volunteers (HbAA). The IH markers evaluated were serum Lactate Dehydrogenase (LDH), total heme, plasma hemoglobin (pHb), and soluble CD163 (sCD163). The ED markers analyzed were plasma von Willebrand factor (VWF:Ag), VWF ristocetin cofactor activity (VWF:RCo) levels, antigen of VWF-cleaving protease (ADAMTS13:Ag), thrombospondin-1, endothelin-1 levels, and ADAMTS13 Activity (ADAMTS13:Act). The levels of VWF:Ag, VWF:RCo, total heme, thrombospondin-1, and endothelin-1 were significantly higher in SCA patients (HbSS and HbSS_HU) compared to HbAA individuals. Also, pHb, LDH, and thrombospondin-1 levels were significantly higher in the HbSS group than in the HbSS_HU group. Contrarily, the levels of sCD163, ADAMTS13:Ag, and ADAMTS13:Act were significantly lower in both groups of SCA patients than HbAA controls, and ADAMTS13:Act levels were significantly lower in HbSS compared to HbSS_HU patients. The higher ADAMTS13 activity levels in those on HU therapy may be attributed to lower pHb and thrombospondin-1 levels as previously shown by in vitro studies that thrombospondin-1 and pHb are bound to VWF. Thus, VWF is restrained from ADAMTS13 activity and cleavage.
Assuntos
Anemia Falciforme/tratamento farmacológico , Endotélio Vascular/fisiopatologia , Hemólise/efeitos dos fármacos , Hidroxiureia/uso terapêutico , Proteína ADAMTS13/sangue , Adolescente , Adulto , Anemia Falciforme/sangue , Antígenos CD/sangue , Antígenos de Diferenciação Mielomonocítica/sangue , Biomarcadores , Estudos Transversais , Endotélio Vascular/efeitos dos fármacos , Feminino , Heme/análise , Hemoglobinas/análise , Humanos , Hidroxiureia/farmacologia , L-Lactato Desidrogenase/sangue , Masculino , Pessoa de Meia-Idade , Proibitinas , Receptores de Superfície Celular/sangue , Trombospondina 1/sangue , Adulto Jovem , Fator de von Willebrand/análiseRESUMO
There are different studies that aim to enhance the production of nisin by Lactococcus lactis since its chemical synthesis is not possible. In this study, glutathione (GSH) and pyruvate, which are known to reduce the oxidative stress of cells, have been shown to trigger the production of nisin at both transcriptional and translational levels in L. lactis cells grown under aerobic condition. Presence of GSH and pyruvate caused more nisin yield than the heme-supplemented medium. Moreover, the expression of genes that encode stress-related enzymes were apparently upregulated in the presence of GSH and pyruvate. It can be concluded that GSH and pyruvate contribute to the defense system of L. lactis cells and so that higher biomass was obtained which in turn enhance nisin production. Antioxidant effect of GSH and pyruvate was known; however, their stimulating effect on nisin production was shown for the first time in this study.
Assuntos
Antibacterianos/biossíntese , Glutationa/metabolismo , Heme/metabolismo , Lactococcus lactis/metabolismo , Nisina/biossíntese , Ácido Pirúvico/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biomassa , Meios de Cultura/análise , Meios de Cultura/metabolismo , Glutationa/análise , Heme/análise , Lactococcus lactis/genética , Lactococcus lactis/crescimento & desenvolvimento , Ácido Pirúvico/análiseRESUMO
Italian salami were sonicated in different times (0, 3, 6 and 9min) using ultrasound bath (US, 25kHz). The effect of sonication on microbial growth (lactic acid bacteria and Micrococcaceae), lipid and protein oxidation, total heme pigments (THP), non heme iron (NHI) and metmyoglobin (MMb) was investigated during processing (0, 2, 15, and 28days) and storage (1, 30, and 120days). US enhanced growth of microorganisms (P<0.05), mainly for the treatment 9min of sonication. The lipid (peroxide value and TBARS) and protein (thiol group) oxidative reactions were accelerated by US (P<0.05) and they should be considered to maintain Italian salami quality. Sonication contributed to maintenance of THP (P<0.05), especially during storage. MMb pigment was not affected by sonication (P>0.05). This study presented some features of US application that could be explored in the manufacture of Italian salami.
Assuntos
Fenômenos Químicos/efeitos da radiação , Microbiologia de Alimentos , Produtos da Carne/efeitos da radiação , Sonicação , Fermentação , Manipulação de Alimentos/métodos , Heme/análise , Heme/efeitos da radiação , Ferro/análise , Ferro/efeitos da radiação , Itália , Lactobacillales/crescimento & desenvolvimento , Lactobacillales/efeitos da radiação , Peroxidação de Lipídeos/efeitos da radiação , Produtos da Carne/análise , Produtos da Carne/microbiologia , Metamioglobina/análise , Metamioglobina/efeitos da radiação , Micrococcaceae/crescimento & desenvolvimento , Micrococcaceae/efeitos da radiação , Oxirredução/efeitos da radiação , Proteínas/química , Proteínas/efeitos da radiação , Substâncias Reativas com Ácido Tiobarbitúrico/análiseRESUMO
Heme iron (Fe) release from alginate beads at in vitro simulated gastrointestinal conditions for potential use as oral heme Fe supplement was studied. Five beads at different ratios of sodium alginate (SA)-to-spray-dried bovine blood cells (SDBC) with weight ratios of 1:1.25, 1:2.5, 1:5, 1:10, and 1:15 (w/w) were prepared. Release characteristics of these beads were investigated at in vitro simulated gastrointestinal conditions. Release media pH strongly influenced the controlled Fe release from the beads. The heme Fe-beads in simulated gastric fluid (pH 2) remained in a shrinkage state and Fe release was low: 25.8, 21.1, 11.6, 12.1, and 12.0 % for 1:1.25, 1:2.5, 1:5, 1:10, and 1:15 ratios, respectively. Proportion and amount of Fe released by 1:1.25 and 1:2.5 ratios was higher than the other ratios. The heme Fe-beads swelled and dissociated in simulated intestinal fluid (pH 6), releasing three-fourths of the Fe in 200 min. The morphology studies showed that Fe release followed formation of pores in the alginate matrix, generating erosion of the beads and complete disintegration after 75 and 200 min of gastric and intestinal incubation, respectively. These results indicate that heme Fe-beads may be useful for oral delivery of heme Fe supplement.
Assuntos
Alginatos/química , Suco Gástrico/química , Heme/análise , Heme/química , Ferro/análise , Ferro/química , Microesferas , Animais , Células Sanguíneas , Bovinos , Suco Gástrico/metabolismo , Ácido Glucurônico/química , Ácidos Hexurônicos/química , Técnicas In VitroRESUMO
A Leishmaniose visceral (LV) apresenta ampla distribuição geográfica e é fatal caso não seja tratada. As manifestações hematológicas são constantes na LV e em casos não tratados os pacientes evoluem à óbito por sangramento maciço ou anemia grave. Neste cenário, mecanismos ligados à morte celular, hemólise, metabolismo do heme e atividade da enzima heme oxigenase podem estar envolvidos na imunopatogênese da LV. A heme oxigenase (HO) tem importantes propriedades regulatórias e está envolvida em processos fisiológicos e patofisiológicos como citoproteção e inflamação. Nesse projeto testamos a hipótese de que a ativação da enzima heme oxigenase-1 (HO-1) favorece a infecção por Leishmania infantum chagasi, principal agente etiológico da LV humana no Brasil e de que mecanismos de morte celular inflamatória induzida por heme estão associados com a resistência ao parasita. Nossas observações nesse trabalho indicam que a enzima HO-1 é induzida em macrófagos durante a infecção por L. chagasi e que a indução farmacológica da HO-1, pela CoPP aumenta a carga parasitária de macrófagos infectados por L. chagasi e reduz a produção de mediadores próinflamatórios. Além disso, a HO-1 favorece um ambiente anti-inflamatório onde prevalece a presença de IL-10...
Visceral leishmaniasis (VL) is a widespread disease and is fatal if left untreated. Hematological manifestations are common in VL and untreated patients evolve to death from massive bleeding and severe anemia. In this scenario, mechanisms related to cell death pathways, hemolysis, heme metabolism and enzymatic activity of heme oxygenase may be involved in the immunopathogenesis of the disease. Heme oxygenase (HO) has important regulatory properties and is involved in patho-physiological processes such as cytoprotection and inflammation. This project tested the hypothesis that heme oxygenase- 1 (HO-1) activation favors Leishmania infantum chagasi infection, the main etiologic agent of human VL in Brazil, we also tested whether heme induced inflammatory cell death pathways are involved in resistance to Leishmania infection. Our observations indicate that HO-1 is induced in macrophages infected with L. infantum chagasi and pharmacological induction for HO-1 by CoPP increases parasite load of infected macrophages and reduces production on inflammatory mediators. In addition, HO-1 contributes to the anti inflammatory pathway that favors L. chagasi replication through a higher IL-10/TNF-α...
Assuntos
Humanos , Heme/análise , Heme/fisiologia , Macrófagos/imunologia , Macrófagos/microbiologia , Macrófagos/parasitologiaRESUMO
A Leishmaniose é uma doença que atinge milhões de pessoas em todo o mundo, sendo endêmica em muitas áreas. A doença pode apresentar diferentes manifestações clínicas, tais como a Leishmaniose Visceral (LV), que é a forma mais grave e letal, caso não seja tratada. As manifestações hematológicas são comumente associadas à LV, onde os mecanismos relacionados à hemólise e a presença do heme livre, podem interferir no comportamento de neutrófilos. Já foi descrito que o heme livre é uma molécula pró-inflamatória, com a capacidade de induzir migração e ativação dos neutrófilos. No entanto, o efeito do heme sobre neutrófilos humanos durante a infecção por Leishmania chagasi, ainda não foi explorado. A nossa hipótese é que o heme induz ativação de neutrófilos humanos favorecendo o processo inflamatório na infecção por L. chagasi. Nossos resultados mostraram que o heme induz a ativação e apoptose em neutrófilos humanos infectados com L. chagasi, seguido pela sobrevivência do parasita. Moléculas presentes na composição do heme, como a protoporfirina IX (PPIX) e o Fe+2, não alteram o status de ativação dos neutrófilos, mas mantêm o aumento do crescimento parasitário como observado em presença do heme. Além disso, o heme e o Fe+2 aumentam a produção do TGF-β e a atividade da SOD. A inibição farmacológica da enzima SOD com dietilditiocarbamato (DETC) reduz a taxa de proliferação da L. chagasi em neutrófilos infectados. Em conjunto, esses dados indicam que o heme e o Fe+2 podem contribuir como fonte nutricional e controlar o ambiente inflamatório, com a indução de TGF-β e SOD, permitindo a sobrevivência da L. chagasi em neutrófilos humanos. Esse estudo poderá abrir novas perspectivas para o entendimento dos mecanismos imunopatogênicos envolvendo neutrófilos e suas implicações no processo inflamatório da LV.
The Leishmaniasis is a disease that affects millions of people worldwide, being endemic in many areas. The disease may present different clinical manifestations, such as Visceral Leishmaniasis (VL), which is the most severe form and fatal if left untreated. The hematologic manifestations are commonly associated with VL, where the mechanisms related to hemolysis and the presence of free heme may modify the behavior of neutrophils. It has been reported that the free heme is a pro-inflammatory molecule with the ability to induce migration and activation of neutrophils. However, the effect of the heme on human neutrophils during infection by Leishmania chagasi yet hasn't been explored. Our hypothesis is that the heme induces activation of human neutrophils favoring the inflammatory process on the infection by L. chagasi. The results showed that the heme induces activation and apoptosis in human neutrophils infected with L. chagasi, followed by parasite survival. Molecules present in the composition of heme, as protoporfirin IX (PPIX) and Fe+2, do not alter the activation status of neutrophils, but maintain the increased parasite growth as viewed in the presence of heme. Furthermore, the heme and Fe+2 increase the production of TGF-β and SOD activity. The pharmacological inhibition of SOD with diethyldithiocarbamate (DETC) reduces the rate of L. chagasi proliferation in infected neutrophils. Together, these data indicate that the heme and Fe+2 may contribute as a nutritional source and control the inflammatory environment with the induction of TGF-β and SOD, allowing the survival of the L. chagasi in human neutrophils. This study may open new perspectives for the understanding of the immunopathogenic mechanisms involving neutrophils and its implications in the inflammatory process of the VL.
Assuntos
Humanos , Heme/análise , Heme , Leishmaniose Visceral/diagnóstico , Leishmaniose Visceral/etiologia , Leishmaniose Visceral/patologia , Leishmaniose Visceral/parasitologia , Leishmaniose Visceral/transmissãoRESUMO
The content and the bioaccessibility of Se, Cu, Zn, Mn, and Fe were determined in unaged and aged meat (14days) from the Psoas major (PM), Gluteus medius (GM) and Longissimus dorsi (LD) muscles of Hereford (H) and Braford breed (B) steers fed pasture. Furthermore, the content of heme-Fe was determined in the same muscles. The H had a lower content of Cu and a higher content of Fe. Also, H had more heme-Fe than B. The bioaccessibility in unaged meat for Se, Cu, Zn, Mn and Fe ranged between 75 and 91%, 30 and 45%, 40 and 68%, 55 and 95%, and 60 and 70%, respectively. After aging, the bioaccessibility for the same minerals ranged between 58 and 80%, 30 and 48%, 40 and 58%, 75 and 95%, and 59 and 70%, respectively. Aging affected negatively the Se content and its bioaccessibility, in the two breeds. Also, the heme-Fe content was negatively affected by aging in all muscles and breeds.
Assuntos
Dieta , Manipulação de Alimentos/métodos , Ferro da Dieta/análise , Carne/análise , Minerais/análise , Músculo Esquelético/química , Oligoelementos/análise , Ração Animal , Animais , Disponibilidade Biológica , Heme/análise , Humanos , Masculino , Carne/classificação , Especificidade da EspécieRESUMO
OBJECTIVES: To refine the reference range for the zinc protoporphyrin-to-heme ratio (ZnPP/H) of preterm infants, we assessed the impact of maternal risk factors on ZnPP/H and evaluated the impact of changes in iron supplementation on iron status. STUDY DESIGN: The reference range for neonatal ZnPP/H was refined using prospective data from 31 reference infants ≤ 35 weeks' postmenstrual age (PMA) plus retrospective data from 51 infants <30 weeks' PMA, and 59 infants 30-40 weeks' PMA. Cord blood and first week of life values were compared when both were available. The impact of maternal risk factors was assessed by examining prospectively collected ZnPP/H from 48 high-risk infants. The effect of changing iron supplementation guidelines was evaluated by retrospective chart review of serial ZnPP/H from 194 infants. RESULTS: Cord ZnPP/H was lower at 30-35 weeks' gestation than at 24-26 weeks' gestation (P = .01). Cord ZnPP/H values from insulin-dependent diabetic mothers were elevated compared with reference values. Changing the iron supplementation protocol was not associated with improved ZnPP/H measurements. CONCLUSIONS: Cord blood and postnatal reference ranges for ZnPP/H are defined. Iron balance depends on a complex interaction of prenatal and postnatal factors.
Assuntos
Heme/análise , Protoporfirinas/sangue , Feminino , Humanos , Recém-Nascido , Recém-Nascido Prematuro , Masculino , Estudos Prospectivos , Fatores de RiscoRESUMO
Cystathionine beta-synthase (CBS) is a homocysteine metabolizing enzyme that contains pyridoxal phosphate (PLP) and a six-coordinate heme cofactor of unknown function. CBS was inactivated by peroxynitrite, the product of nitric oxide and superoxide radicals. The IC(50) was approximately 150microM for 5microM ferric CBS. Stopped-flow kinetics and competition experiments showed a direct reaction with a second-order rate constant of (2.4-5.0)x10(4)M(-1)s(-1) (pH 7.4, 37 degrees C). The radicals derived from peroxynitrite, nitrogen dioxide and carbonate radical, also inactivated CBS. Exposure to peroxynitrite did not modify bound PLP but led to nitration of Trp208, Trp43 and Tyr223 and alterations in the heme environment including loss of thiolate coordination, conversion to high-spin and bleaching, with no detectable formation of oxo-ferryl compounds nor promotion of one-electron processes. This study demonstrates the susceptibility of CBS to reactive oxygen/nitrogen species, with potential relevance to hyperhomocysteinemia, a risk factor for cardiovascular diseases.
Assuntos
Cistationina beta-Sintase/metabolismo , Ácido Peroxinitroso/farmacologia , Dióxido de Carbono/farmacologia , Cromatografia Líquida de Alta Pressão , Cistationina beta-Sintase/química , Cistationina beta-Sintase/genética , Espectroscopia de Ressonância de Spin Eletrônica , Ativação Enzimática/efeitos dos fármacos , Deleção de Genes , Heme/análise , Heme/metabolismo , Humanos , Cinética , Manitol/farmacologia , Espectrometria de Massas , Ácido Nítrico/metabolismo , Ácido Peroxinitroso/metabolismo , Fenilacetatos/farmacologia , Multimerização Proteica , Estrutura Quaternária de Proteína , Espectrofotometria Ultravioleta , Compostos de Sulfidrila/metabolismoRESUMO
To determine the content of total iron (TFe) and heme iron (HeFe) in major cuts of meat and principal viscera of bovine origin. 55Fe (30 mCi) was injected into two 4-month-old calves. Triplicate samples of the 12 basic American cuts of meat and major viscera were obtained from each specimen. Samples were acid digested and their iron content was read by atomic absorption spectrophotometry. Duplicate samples of the basic cuts of meat and major viscera were analyzed to determine the concentration of 55Fe using a double isotopic technique. The mean and standard deviation of TFe for all cuts was 1.4 ± 0.3 mg/100 g of meat. The mean TFe for organs was (per mg/100 g): 0.9 ± 0.1 brain, 3.0 ± 0.05 kidney, 3.2 ± 0.04 heart, 5.7 ± 0.2 lung, 6.0 ± 0.1 liver, and 31.2 ± 0.4 spleen. HeFe was 64% of TFe in meat and 72.8% in spleen, 53.8% in lung, 35.7% in brain, 35.0% in kidney, 27.3% in heart, and only 13.6% in liver. Blood contained 85.5% of the radioisotope and only 1.4% was found in muscle and 1.6% was found in viscera. Results suggest that bovine cuts of meat have a low variation in TFe and that HeFe comprises more than 60% of TFe.