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1.
Biochem Biophys Res Commun ; 676: 171-181, 2023 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-37517220

RESUMO

A novel carbonic anhydrase II (CA II) from erythrocytes of camel (Camelus dromedarius) was purified to homogeneity using affinity chromatography and biochemically characterized. Specific activity of 140.88 U/mg was obtained with 745.17-fold purification and 25.37% yield. The enzyme was a monomer with a lower molecular weight (25 kDa) and lower Zn content (0.50 mol of Zn per mol of protein). The enzyme showed higher optimum temperature (70 °C) and pH (pH 9.0), moreover, it was stable at higher temperatures and strongly alkaline pH as judged by thermodynamic parameters (Ea, kd, Ed, t1/2, D-value, Z-value, ΔH, ΔG and ΔS). The enzyme was inhibited by cations (Al3+, Ca2+, Cd2+, Co2+, Cr3+, Cu2+, Fe3+, Ni2+, Mg2+ and Zn2+) as well as by anions (Br‾, CH3COO‾, ClO4‾, CN‾, F‾, HCO3‾, I‾, N3‾, NO3‾ and SCN‾), some anions (C6H5O73-, CO32-, SeO3‾ and SO42-) does not affect enzyme activity. Effect of various chemicals on enzyme activity was also investigated. Km, Vmax, kcat and kcat/Km values for 4-NPA were found to be 1.74 mM, 0.0093 U/mL, 0,0039 s-1 and 0,0023 s-1 mM-1, respectively. With these interesting biochemical properties, camel CA II represents promising candidate for harsh industrial applications, in particular, for a successful biomimetic CO2 sequestration process.

2.
Prep Biochem Biotechnol ; 53(6): 610-621, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36074915

RESUMO

Camel is continually exposed to stressful desert environment that enhances generation of reactive oxygen species, including hydrogen peroxide (H2O2). Catalase plays an important role in detoxification of H2O2. A highly active catalase from camel kidney was purified to homogeneity, with a specific activity of 1,774,392 U/mg protein, using ion exchange and metal chelate affinity chromatography. The molecular weight of the enzyme was 268 kDa consisting of four identical subunits of 63 kDa. The enzyme showed higher optimum temperature (45 °C) and higher activation energy (4.37 kJ mol-1). The thermodynamic parameters, ΔH, ΔG and ΔS, were determined. The effect of various metal ions and chemicals on enzyme activity was investigated. Km, Vmax, kcat and kcat/Km values for H2O2 were found to be 46 mM, 10,715,045 U/mg, 48,265,968 s-1 and 2,966,562 s-1 mM-1, respectively. Camel kidney catalase displayed higher affinity efficiency for H2O2 and can protect reduced glutathione (GSH) from oxidation by H2O2. Sodium azide was found to be a noncompetitive inhibitor of enzyme with Ki and IC50 of 17.88 µM and 20.94 µM, respectively. Camel catalase showed unique biochemical properties. Interestingly, camel catalase can protect molecules (GSH) and organ functions (kidney) from the toxic effects of H2O2 induced by stressful desert environment.


Assuntos
Camelus , Peróxido de Hidrogênio , Animais , Catalase/química , Camelus/metabolismo , Peróxido de Hidrogênio/química , Termodinâmica , Metais , Rim/metabolismo , Concentração de Íons de Hidrogênio
3.
Molecules ; 28(15)2023 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-37570894

RESUMO

Recently, the study of the protective powers of medicinal plants has become the focus of several studies. Attention has been focused on the identification of new molecules with antioxidant and chelating properties to counter reactive oxygen species (ROS) involved as key elements in several pathologies. Considerable attention is given to argan oil (AO) and olive oil (OO) due to their particular composition and preventive properties. Our study aimed to determine the content of AO and OO on phenolic compounds, chlorophylls, and carotenoid pigments and their antioxidant potential by FRAP and DPPH tests. Thus, several metallic elements can induce oxidative stress, as a consequence of the formation of ROS. Iron is one of these metal ions, which participates in the generation of free radicals, especially OH from H2O2 via the Fenton reaction, initiating oxidative stress. To study the antioxidant potential of AO and OO, we evaluated their preventives effects against oxidative stress induced by ferrous sulfate (FeSO4) in the protozoan Tetrahymena pyriformis and mice. Then, we evaluated the activities of the enzymatic (superoxide dismutase (SOD), glutathione peroxidase (GPx)) and metabolite markers (lipid peroxidation (MDA) and glutathione (GSH)) of the antioxidant balance. The results of the antioxidant compounds show that both oils contain phenolic compounds and pigments. Moreover, AO and OO exhibit antioxidant potential across FRAP and DPPH assays. On the other hand, the results in Tetrahymena pyriformis and mice show a variation in the level of iron-changed SOD and GPx activities and MDA and GSH levels. By contrast, treating Tetrahymena pyriformis and mice with argan and olive oils shows significant prevention in the SOD and GPx activities. These results reveal that the iron-changed ROS imbalance can be counteracted by AO and OO, which is probably related to their composition, especially their high content of polyphenols, sterols, and tocopherols, which is underlined by their antioxidant activities.


Assuntos
Antioxidantes , Ferro , Camundongos , Animais , Antioxidantes/farmacologia , Antioxidantes/metabolismo , Azeite de Oliva/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Ferro/farmacologia , Peróxido de Hidrogênio/farmacologia , Óleos de Plantas/farmacologia , Óleos de Plantas/química , Estresse Oxidativo , Peroxidação de Lipídeos , Glutationa/metabolismo , Fenóis/farmacologia , Superóxido Dismutase/metabolismo
4.
Bioorg Chem ; 86: 428-436, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30771689

RESUMO

A novel copper, zinc superoxide dismutase (CuZnSOD) was purified to homogeneity from the liver of an animal well adapted to the stressful living conditions of the desert, the camel (Camelus dromedarius). The biochemical properties of camel liver CuZnSOD were examined. The purified enzyme had a native molecular weight of 28 kDa, as judged by gel filtration chromatography, and showed a single band at 27 kDa on SDS-PAGE, indicating that it is a monomeric protein. Optimal activity of the purified enzyme occurred at 43 °C and pH 6.0, and the activation energy was 1.42 kJ/mol. CuZnSOD activity was strongly inhibited by ß-ME, DTT, H2O2 and SDS and slightly inhibited by EDTA, NaN3 and PMSF. Al3+, Ca2+, Cd2+, Mg2+ and Zn2+ stimulated CuZnSOD activity, whereas Ba2+, Co2+, Fe2+ and Ni2+ inhibited it. The purified enzyme contained 0.010 µg of Cu and 0.69 µg of Zn per mg of protein. Km, Vmax, kcat and kcat/Km values for NBT and riboflavin were 16.27 and 0.16 µM, 20.85 and 21.54 U/mg, 9.65 and 9.97 s-1, and 0.59 and 62.33 s-1 µM-1, respectively. Camel liver CuZnSOD exhibited unique biochemical properties compared to those of other CuZnSODs, including lower molecular weight with a monomeric structure, higher optimum temperature, very low Ea, very low optimum pH, very low contents of Cu and Zn, and higher affinity, turnover number and catalytic efficiency for riboflavin. These unique properties of camel liver CuZnSOD might be related to the ability of this animal to inhabit stressful desert conditions.


Assuntos
Antioxidantes/metabolismo , Cobre/metabolismo , Fígado/enzimologia , Superóxido Dismutase/metabolismo , Zinco/metabolismo , Animais , Antioxidantes/química , Antioxidantes/isolamento & purificação , Camelus , Cobre/química , Cobre/isolamento & purificação , Relação Dose-Resposta a Droga , Cinética , Estrutura Molecular , Relação Estrutura-Atividade , Superóxido Dismutase/química , Superóxido Dismutase/isolamento & purificação , Zinco/química , Zinco/isolamento & purificação
5.
Int J Mol Sci ; 18(10)2017 Oct 19.
Artigo em Inglês | MEDLINE | ID: mdl-29048364

RESUMO

Sepsis causes severe dysregulation of organ functions, via the development of oxidative stress and inflammation. These pathophysiological mechanisms are mimicked in mice injected with bacterial lipopolysaccharide (LPS). Here, protective properties of argan oil against LPS-induced oxidative stress and inflammation are explored in the murine model. Mice received standard chow, supplemented with argan oil (AO) or olive oil (OO) for 25 days, before septic shock was provoked with a single intraperitoneal injection of LPS, 16 hours prior to animal sacrifice. In addition to a rise in oxidative stress and inflammatory markers, injected LPS also caused hepatotoxicity, accompanied by hyperglycemia, hypercholesterolemia and hyperuremia. These LPS-associated toxic effects were blunted by AO pretreatment, as corroborated by normal plasma parameters and cell stress markers (glutathione: GSH) and antioxidant enzymology (catalase, CAT; superoxide dismutase, SOD and glutathione peroxidase, GPx). Hematoxylin-eosin staining revealed that AO can protect against acute liver injury, maintaining a normal status, which is pointed out by absent or reduced LPS-induced hepatic damage markers (i.e., alanine aminotransferase (ALT) and aspartate transaminase (AST)). Our work also indicated that AO displayed anti-inflammatory activity, due to down-regulations of genes encoding pro-inflammatory cytokines Interleukin-6 (IL-6) and Tumor Necrosis Factor-α (TNF-α) and in up-regulations of the expression of anti-inflammatory genes encoding Interleukin-4 (IL-4) and Interleukin-10 (IL-10). OO provided animals with similar, though less extensive, protective changes. Collectively our work adds compelling evidence to the protective mechanisms of AO against LPS-induced liver injury and hence therapeutic potentialities, in regard to the management of human sepsis. Activations of IL-4/Peroxisome Proliferator-Activated Receptors (IL-4/PPARs) signaling and, under LPS, an anti-inflammatory IL-10/Liver X Receptor (IL-10/LXR) route, obviously indicated the high potency and plasticity of the anti-inflammatory properties of argan oil.


Assuntos
Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Hepatopatias/tratamento farmacológico , Fígado/efeitos dos fármacos , Azeite de Oliva/farmacologia , Estresse Oxidativo , Óleos de Plantas/farmacologia , Animais , Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/uso terapêutico , Antioxidantes/administração & dosagem , Antioxidantes/uso terapêutico , Suplementos Nutricionais , Lipopolissacarídeos/toxicidade , Fígado/metabolismo , Hepatopatias/etiologia , Hepatopatias/prevenção & controle , Camundongos , Azeite de Oliva/administração & dosagem , Azeite de Oliva/uso terapêutico , Óleos de Plantas/administração & dosagem , Óleos de Plantas/uso terapêutico
6.
Theriogenology ; 215: 234-240, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38100995

RESUMO

This research sought to purify C-phycocyanin (C-PC) from Spirulina platensis and investigate its potential in enhancing the quality parameters and in vivo fertility of ram semen subjected to cooled storage at 5 °C, when using a skim milk (SM) based semen extender. The purification process of C-PC involved cold maceration, pre-purification using chitosan and activated charcoal, followed by purification through aqueous two-phase extraction (ATPE) and ion-exchange chromatography. Afterward, fifty ejaculates were collected from 4 fertile Boujaâd rams and extended using the SM extender at 37 °C, enriched with 0 µg/mL (control), 1.2 µg/mL, 2.4 µg/mL, 3.6 µg/mL, or 4.8 µg/mL of C-PC. The diluted semen was subsequently cooled to 5 °C using a controlled cooling process, with a gradual cooling rate of approximately 0.5 °C per minute, and its quality parameters were evaluated after 0, 4, 8, and 24 h of cooling storage. Then, its fertilization ability after 4 h of cooling storage was evaluated using artificial insemination. The adopted purification process yielded a grade analytical purity of 4.06. Additionally, semen extended in SM with a 2.4 µg/mL C-PC supplement displayed significant (P < 0.0001) enhancement in total motility, progressive motility, curvilinear velocity, straight-line velocity, average path velocity, viability and lipid peroxidation of ram semen at 0, 4, 8, and 24 h of cooling storage. These improvements were observed in direct comparison to both the control group and the other C-PC concentrations. Regarding fertility rates, semen extended in SM with a 2.4 µg/mL C-PC recorded a 76 % rate, a notable increment from the 63 % observed in ewes inseminated by semen extended in SM alone, although the difference was not statistically significant (p > 0.05). These findings underscore the promising potential of C-PC as a natural supplement for enhancing semen quality, warranting further investigations.


Assuntos
Análise do Sêmen , Preservação do Sêmen , Ovinos , Animais , Masculino , Feminino , Análise do Sêmen/veterinária , Ficocianina/farmacologia , Preservação do Sêmen/veterinária , Preservação do Sêmen/métodos , Motilidade dos Espermatozoides , Fertilidade , Sêmen , Carneiro Doméstico , Espermatozoides
7.
Front Pharmacol ; 15: 1345340, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38455958

RESUMO

This study assessed the medicinal properties of Euphorbia resinifera O. Berg (E. resinifera) and Euphorbia officinarum subsp echinus (Hook.f. and Coss.) Vindt (Euphorbia echinus, known for their pharmaceutical benefits. Extracts from their flowers, stems, propolis, and honey were examined for phenolic content, antioxidant, anti-inflammatory, and antibacterial activities. Total phenolic content (TPC), total flavonoid content (TFC), and total condensed tannin (TCC) were determined using specific methods. Antioxidant potential was assessed through various tests including DPPH, FRAP, ABTS, and Total antioxidant capacity. Anti-inflammatory effects were evaluated using phenol-induced ear edema in rats, while antibacterial activity was measured against Gram-positive (Staphylococcus aureus ATCC 6538) and Gram-negative (E. coli ATCC 10536) bacteria. Among the extracts, the aqueous propolis extract of E. resinifera demonstrated exceptional antioxidant capabilities, with low IC50 values for DPPH (0.07 ± 0.00 mg/mL) and ABTS (0.13 ± 0.00 mg/mL), as well as high TAC (176.72 ± 0.18 mg AA/mg extract) and FRAP (86.45 ± 1.45 mg AA/mg extract) values. Furthermore, the anti-inflammatory effect of E. resinifera propolis extracts surpassed that of indomethacin, yielding edema percentages of 3.92% and 11.33% for the aqueous and ethanolic extracts, respectively. Microbiological results indicated that the aqueous extract of E. resinifera flower exhibited the most potent inhibitory action against S. aureus, with an inhibition zone diameter (IZD) of 21.0 ± 0.00 mm and a minimum inhibitory concentration (MIC) of 3.125 mg/mL. Additionally, only E. resinifera honey displayed the ability to inhibit E. coli growth, with an inhibition zone diameter of 09.30 ± 0.03 mm and a MIC of 0.0433 mg/mL.

8.
Biomedicines ; 11(4)2023 Apr 18.
Artigo em Inglês | MEDLINE | ID: mdl-37189816

RESUMO

This study aimed to determine the size and distribution of LDL and HDL particles in North African acute coronary syndrome (ACS) patients and to compare the level of small dense LDL (sdLDL) to other markers used in cardiovascular risk prediction. METHODS: A total of 205 ACS patients and 100 healthy control subjects were enrolled. LDL particle size and LDL and HDL subclass distributions were measured using Quantimetric Lipoprint® linear polyacrylamide gel electrophoresis. Lipid ratios (total cholesterol, LDL cholesterol, non-HDL cholesterol, and HDL cholesterol) were determined to calculate the atherogenic index of plasma (AIP), the atherogenic coefficient (AC), Castelli's Risk-I (CR-I), and Castelli's Risk-II (CR-II). Receiver operating characteristic (ROC) curve analyses and area under the curve (AUC) were used to assess the predictive value of sdLDL as a marker for cardiovascular disease. RESULTS: The ACS patients, compared to the healthy control subjects, displayed an alteration of LDL particle distribution, with a significant increase in sdLDL serum concentrations (0.303 ± 0.478 mmol/L vs. 0.0225 ± 0.043 mmol/L, respectively, p < 0.001). The sdLDL levels had a high discrimination accuracy [AUC = 0.847 ± 0.0353 (95% CI 0.778 to 0.916, p < 0.0001)]. The best predictive cutoff value of ACS determined with the maximum Youden index (J) [(sensitivity + specificity) - 1 = 0.60] was 0.038 mmol/L. A Spearman correlation analysis showed that sdLDL levels were moderately but significantly and positively correlated with AC and CR-I (r = 0.37, p < 0.001) and weakly but significantly correlated with PAI and CR-II; r = 0.32 (p < 0.001) and r = 0.30 (p < 0.008), respectively. The subclass distribution of HDL particles from ACS patients was also altered, with a decrease in large HDL particles and an increase in small HDL particles compared to HDL from healthy control subjects. CONCLUSION: Due to their high atherogenicity, sdLDL levels could be used as a valuable marker for the prediction cardiovascular events.

9.
Toxicology ; 465: 153025, 2022 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-34748892

RESUMO

Fluoride is a natural element widely distributed in the environment and plays an important role in the growth of humans and animals. However, in many species, high concentrations of fluoride induce several problems, such as dental, skeletal, and non-skeletal fluorosis. Sheep living in endemic areas are sensitive to the chronic toxicity of fluoride, and they have been found to suffer not only from teeth and bone problems but also from other organs. Studies indicating the chronic harmful effects of fluoride on teeth, bones, blood biochemical parameters, kidney, liver, heart, reproductive system and growth in sheep have been clearly summarized in this review. Besides, this work also includes updated progress in terms of prevention or reduction of fluoride toxicity in this species.


Assuntos
Antioxidantes/administração & dosagem , Doenças Ósseas/veterinária , Suplementos Nutricionais , Doenças Endêmicas/veterinária , Flúor/toxicidade , Fluorose Dentária/veterinária , Minerais/administração & dosagem , Doenças dos Ovinos/prevenção & controle , Ração Animal , Animais , Doenças Ósseas/induzido quimicamente , Doenças Ósseas/diagnóstico , Doenças Ósseas/prevenção & controle , Doenças Endêmicas/prevenção & controle , Fluorose Dentária/diagnóstico , Fluorose Dentária/etiologia , Fluorose Dentária/prevenção & controle , Fatores de Proteção , Medição de Risco , Fatores de Risco , Ovinos , Doenças dos Ovinos/induzido quimicamente , Doenças dos Ovinos/diagnóstico
10.
Pharmaceuticals (Basel) ; 15(4)2022 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-35455460

RESUMO

During sepsis, the imbalance between oxidative insult and body antioxidant response causes the dysfunction of organs, including the brain and liver. Exposing mice to bacterial lipopolysaccharides (LPS) results in a similar pathophysiological outcome. The protection offered by argan oil was studied against LPS-induced oxidative stress, dysregulation of peroxisomal antioxidants, and ß-oxidation activities in the brain and liver. In a short-term LPS treatment, lipid peroxidation (malonaldehyde assay) increased in the brain and liver with upregulations of proinflammatory tumor necrosis factor (Tnf)-α and anti-inflammatory interleukin (Il)-10 genes, especially in the liver. Although exposure to olive oil (OO), colza oil (CO), and argan oil (AO) prevented LPS-induced lipid peroxidation in the brain and liver, only AO exposure protected against liver inflammation. Remarkably, only exposure to AO prevented LPS-dependent glutathione (GSH) dysregulation in the brain and liver. Furthermore, exposure to AO increased more efficiently than OO and CO in both organs, peroxisomal antioxidant capacity via induction of catalase (Cat) gene, protein and activity expression levels, and superoxide dismutase (Sod1) mRNA and activity levels. Interestingly, LPS decreased protein levels of the peroxisomal fatty acid-ATP binding cassette (ABC) transporters, ABCD1 and ABCD2, and increased acyl-CoA oxidase 1 (ACOX1) protein expression. Moreover, these LPS effects were attenuated for ABCD1 and ACOX1 in the brain of mice pretreated with AO. Our data collectively highlight the protective effects of AO against early oxidative stress caused by LPS in the brain and liver and their reliance on the preservation of peroxisomal functions, including antioxidant and ß-oxidation activities, making AO a promising candidate for the prevention and management of sepsis.

11.
Int J Biol Macromol ; 160: 991-999, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-32454111

RESUMO

Acid phosphatase (ACP) plays an important role in regulating phosphate nutrition in plants. Herein, for the first time, a novel ACP from Opuntia megacantha Salm-Dyck cladodes was purified to homogeneity and biochemically characterized. Specific activity of 8.78 U/mg was obtained with 11.29-fold purification and 15% yield. ACP was purified as monomer with molecular weight of 44 kDa as determined by SDS-PAGE under denaturing and nondenaturing conditions. Optimum pH and temperature for ACP activity was 5.5 and 60 °C, respectively. Thermodynamic parameters (Ea, ΔH, ΔG and ΔS) were also determined. ACP activity was stimulated by Ca2+, strongly inhibited by Cu2+ and Fe3+, and moderately inhibited by Mg2+ and Zn2+. Br-, CN-, F-, I- and N3- weakly inhibited ACP activity, where more than 70% of enzyme activity was remained at 5 mM. In addition, effect of ß-ME, Cys, DTT, EDTA, H2O2, PMSF, SDS and TX-100 on ACP activity was investigated. km, Vmax, kcat and kcat/km of ACP for p-NPP were found to be 0.09 mM, 2.75 U/mL, 9.60 s-1 and 106.67 s-1 mM-1, respectively. The biochemical properties of ACP from Opuntia megacantha Salm-Dyck cladodes provide novel features with other plant ACPs and basic knowledge of ACP in Opuntia species.


Assuntos
Fosfatase Ácida/química , Fosfatase Ácida/isolamento & purificação , Opuntia/enzimologia , Fenômenos Químicos , Ativação Enzimática , Concentração de Íons de Hidrogênio , Cinética , Peso Molecular , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Termodinâmica
12.
Int J Food Sci ; 2020: 8885889, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33029486

RESUMO

Pomegranate (Punica granatum L) is widely cultivated in the Mediterranean countries especially in Morocco. Pomegranate peel and seed contain considerable amounts of phenolic compounds with antioxidant activity. The aim of the present study was to phytochemically characterize the pomegranate peels and seeds obtained from three Moroccan provinces, using UHPLC-DAD. In addition, total phenolic content (TPC), total flavonoid contents (TFC), and metal chelating of pomegranate peel were also evaluated. The results showed that pomegranate peel possesses the highest phenolic (TPC: 224.39 mg GAE/g dw) and flavonoid (TFC: 62.64 mg rutin/g dw) contents. Punicalagin-ß and punicalagin-α, are the abundant compounds found in peel: 216.36 ± 9.94 mg/g, 154.94 ± 5.21 mg/g, respectively. Pomegranate peels showed significantly (p < 0.05) high antioxidant activity 1-diphenyl-2-picrylhydrazyl (DPPH) EC50: 42.71 ± 0.04 µg/mL, 2.2'-Azino-bis(3-Ethylbenzothiazoline-6-Sulfonic Acid) (ABTS) EC50: 62.15 ± 0.01 µg/mL), and chelating activity (FRAP 1.85 ± 0.00 mg ascorbic acid equivalents/100 g, Fe2+: 2.52 ± 0.01 µmol EDTA equivalents/g dw) compared to seeds. A positive correlation between antioxidant activity and total phenolic was found. According to achieved results, high antioxidant capacity of pomegranate extracts, especially peel, shed light to further use as natural food preservatives. Pomegranate peel could be used for the fortification of food with fiber by introducing it in dietary, as well as in health applications due to its higher antioxidant capacity.

13.
Environ Toxicol ; 23(4): 539-47, 2008 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-18214938

RESUMO

Industrial processing of phosphates generates chemical wastes which are, without any treatment, discharged directly into the Atlantic Ocean at Jorf Lasfar (JL), located 120 km south of Casablanca (Morocco) were shellfish are also collected by people without any control. Marine bivalves concentrate these pollutants by filtration and serve as vectors in human's exposure. The objective of this study was to test and compare in vitro on human intestinal cells (Caco-2) the cytotoxicity and genotoxicity of mussels (Mytilus galloprovincialis) extracts (either hydrophilic or lipophilic) collected at two coastal sites; JL (neighboring a phosphate processing plat-form) and Oualidia (OL) (a vegetable growing area) located 160 km south of Casablanca (i.e. 40 km south of JL). Using Caco-2 cells, the following end-points have been evaluated, cytotoxicity as measured by MTS test, inhibition of cellular macromolecules syntheses (DNA and protein) and genotoxicity evaluated by DNA fragmentation in agarose gel electrophoresis. The results indicated, that hydrophilic and lipophilic OL mussels extracts are cytotoxic and inhibit cellular macromolecules syntheses. Moreover these extracts damage the DNA in Caco-2 cells. The lipophilic JL mussels extract is cytotoxic, inhibits cellular macromolecules syntheses, and damages the DNA in Caco-2 cells whereas the hydrophilic extract of JL mussels fails to inhibit protein synthesis and does not damage the DNA. This extract rather enhances protein synthesis, suggesting possible metallothioneins induction by metal ions. Altogether these in vitro data indicate that mussels collected from OL could be more harmful than those from JL even though the later is closer to the pollution site than OL. Nevertheless consumption of mussels from all these areas may present a risk for humans. Epidemiological studies will be needed for global risk assessment in humans living in these areas especially those consuming see food regularly.


Assuntos
Bivalves/química , Citotoxinas/toxicidade , Células Epiteliais/efeitos dos fármacos , Mutagênicos/toxicidade , Animais , Oceano Atlântico , Células CACO-2 , Citotoxinas/química , Relação Dose-Resposta a Droga , Humanos , Mucosa Intestinal/citologia , Metais/análise , Metais/toxicidade , Marrocos , Mutagênicos/química
14.
Anim Reprod Sci ; 197: 40-47, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30143278

RESUMO

The present research was conducted to evaluate the effects of season and age on scrotal circumference (SC), semen characteristics, semen composition, and sperm motility during liquid storage in Boujaâd rams. Semen was collected throughout 1 year from April 2014 to March 2015. Two ram groups, namely older rams including animals aged 5.5 to 6 years of age, and younger rams including animals aged 2.5 to 3 years of age were used. Scrotal circumferences, semen characteristics, and some biochemical variables from semen were measured. After collection and evaluation, the semen was extended in skim milk (SM) at 15 °C to a concentration of 0.8 × 109 spermatozoa/ml. Thereafter, samples were evaluated at different storage times (0, 8, and 24 h). The results indicate that older rams had greater values of SC, lesser semen quality, protein, lipid, and cholesterol concentrations than younger rams. Nevertheless, for both groups there were the least values for semen quality and semen composition in winter. The semen collected from younger rams had greater motility after liquid storage compared with that of the older rams. The least semen quality after liquid storage was recorded in the winter compared with the other seasons of the year for both age groups. In conclusion, results of the present study indicate that older Boujaâd sheep have lesser semen quality, different semen composition than younger rams after liquid storage and during different seasons of the year. Furthermore, there was a strong correlation between semen composition and motility of stored ram semen.


Assuntos
Envelhecimento/fisiologia , Preservação do Sêmen/veterinária , Sêmen/fisiologia , Ovinos/fisiologia , Animais , Masculino , Estações do Ano , Análise do Sêmen , Motilidade dos Espermatozoides , Espermatozoides
15.
Artigo em Inglês | MEDLINE | ID: mdl-29100758

RESUMO

Climate change and increasing temperatures are global concerns. Camel (Camelus dromedarius) lives most of its life under high environmental stress in the desert and represent ideal model for studying desert adaptation among mammals. Catalase plays a key role in protecting cells against oxidative stress. For the first time, catalase from camel liver was purified to homogeneity by zinc chelate affinity chromatography using pH gradient elution, a better separation was obtained. A purification fold of 201.81 with 1.17% yield and a high specific activity of 1132539.37U/mg were obtained. The native enzyme had a molecular weight of 268kDa and was composed of four subunits of equal size (65kDa). The enzyme showed optimal activity at a temperature of 45°C and pH 7.2. Thiol reagents, ß-Mercaptoethanol and D,L-Dithiothreitol, inhibited the enzyme activity. The enzyme was inhibited by Al3+, Cd2+ and Mg2+, whereas Ca2+, Co2+ and Ni2+ stimulated the catalase activity. Reduced glutathione has no effect on catalase activity. The Km and Vmax of the enzyme for hydrogen peroxide were 37.31mM and 6185157U/mg, respectively. Sodium azide inhibited the enzyme noncompetitively with Ki value of 14.43µM, the IC50 was found to be 16.71µM. The properties of camel catalase were different comparing to those of mammalian species. Relatively higher molecular weight, higher optimum temperature, protection of reduced glutathione from hydrogen peroxide oxidation and higher affinity for hydrogen peroxide and sodium azide, these could be explained by the fact that camel is able to live in the intense environmental stress in the desert.


Assuntos
Catalase/química , Catalase/isolamento & purificação , Cromatografia de Afinidade/métodos , Fígado/enzimologia , Animais , Camelus , Catalase/antagonistas & inibidores , Catalase/metabolismo , Ácido Edético , Inibidores Enzimáticos , Concentração de Íons de Hidrogênio , Dodecilsulfato de Sódio , Temperatura
16.
PLoS One ; 10(8): e0133719, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26241956

RESUMO

The purpose of the present study was to investigate the distribution of PON1 Q192R and L55M polymorphisms and activities in a North African population and to determine their association with cardiovascular complications. The prevalence of the QQ, QR, RR, LL, LM, and MM genotypes in the study population was 55.4%, 34.09%, 9.83%, 41.97%, 48.20%, and 9.83% respectively. The Q, R, L, and M alleles had a gene frequency of 0.755, 0.245, 0.67, and 0.33, respectively. The PON1 192 RR genotype was significantly more prevalent among ACS patients than among healthy subjects. There was a 4.33-fold increase in the risk of ACS in subjects presenting the PON1 192 RR genotype compared to those with the QQ genotype (OR=4.33; 95% CI=1.27-17.7). There was a significantly different distribution of PON1 L55M in the ACS patient groups (UA, STEMI, NSTEMI). Moreover, individuals presenting the PON1 55MM genotype present a higher risk for ACS than those with LL genotype (OR=3.69; 95% CI=1.61-11.80). Paraoxonase activities were significantly lower in coronary patients than in healthy subjects. The decrease in PON1 activity was inversely correlated with the number of concomitant risk factors for CVD (r=0.57, p<0.0001). The results of the present study suggested that the PON1 R and M alleles may play a role in the pathogenesis of cardiac ischemia in our North African population and that a decrease in PON1 activity may be a valuable marker for monitoring the development of the atherosclerosis process and the associated cardiovascular complications.


Assuntos
Síndrome Coronariana Aguda/genética , Arildialquilfosfatase/genética , Polimorfismo Genético , Síndrome Coronariana Aguda/sangue , Síndrome Coronariana Aguda/epidemiologia , Alelos , Substituição de Aminoácidos , Glicemia/análise , Pressão Sanguínea , Índice de Massa Corporal , Feminino , Frequência do Gene , Estudos de Associação Genética , Predisposição Genética para Doença , Genótipo , Humanos , Lipídeos/sangue , Masculino , Pessoa de Meia-Idade , Marrocos , Estresse Oxidativo , Risco , Fatores de Risco
17.
Anim Reprod Sci ; 160: 57-67, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26235670

RESUMO

Due to its high antioxidant content, the argan oil could play a beneficial role in liquid storage of ram semen. The aim of this study was to investigate effects of different concentration of argan oil (ARO) on spermatologic parameters, lipid peroxidation and DNA fragmentation during liquid storage of ram semen until 48 h. Also effects of extenders and temperature on same parameters were assessed. For these aims, semen samples were collected from Boujaâd rams, extended with Tris egg yolk or skim milk extenders without (control) or supplemented with different concentrations of ARO (1%, 2%, 5% and 10% v/v) at a final concentration of 0.8 × 10(9) sperm/mL and stored until 48 h at 5 °C or 15 °C. The sperm quality assessments were performed at different intervals during storage (0, 8, 24 and 48 h). Sperm progressive motility started to decrease after 8h of storage in all temperatures--extenders combinations and dropped steadily during the 8-48 h interval. However, sperm viability, progressive motility and membrane integrity were markedly higher in ARO groups (especially in 1% in Tris and 5% in skim milk) until 24h and 48 h storage at both temperatures compared to controls. The argan oil also decreased the level of spontaneous and induced malondialdehyde (MDA) and the sperm DNA fragmentation until 48 h storage. In conclusion, it was determined that addition of argan oil to conventional extenders may improve the quality of ram semen during liquid storage in different temperatures.


Assuntos
Leite/química , Óleos de Plantas/farmacologia , Preservação do Sêmen/veterinária , Sêmen/efeitos dos fármacos , Trometamina/farmacologia , Animais , Membrana Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Gema de Ovo , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Óleos de Plantas/administração & dosagem , Ovinos
18.
Clin Biochem ; 47(18): 318-25, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25218815

RESUMO

OBJECTIVE: The functionality of HDL has been suggested as an important factor in the prevention of cardiovascular and coronary artery diseases. The objective of the present study was to investigate the functionality of HDL and the factors that may affect the anti-atherogenic properties of HDL in ACS patients. METHODS AND RESULTS: One hundred healthy subjects and 205 ACS patients were recruited. HDL functionality was evaluated by measuring their capacity to mediate cholesterol efflux from J774 macrophages. Oxidative stress status was determined by measuring plasma malondialdehyde (MDA), protein carbonyl, and vitamin E levels by HPLC. The PON1 Q192R polymorphism status and PON1 paraoxonase and arylesterase activities of the healthy subjects and ACS patients were also determined. The HDL of ACS patients displayed a limited capacity to mediate cholesterol efflux, especially via the ABCA1-pathway. MDA (7.06±0.29 µM) and protein carbonyl (9.29±0.26 µM) levels were significantly higher in ACS patients than in healthy subjects (2.29±0.21 µM and 3.07±0.17 µM, respectively, p<0.0001), while α- and γ-tocopherol (vitamin E) levels in ACS patients were 8-fold (p<0.001) and 2-fold (p<0.05) lower than in healthy subjects. Paraoxonase, arylesterase and HDL-corrected PON1 activities (PON1 activity/HDL ratio) were significantly lower in ACS patients. Logistic regression analyses showed that high PON1 paraoxonase and arylesterase activities had a significant protective effect (OR=0.413, CI 0.289-0.590, p<0.001; OR=0.232 CI 0.107-0.499, p<0.001, respectively) even when adjusted for HDL level, age, BMI, and PON1 polymorphism. CONCLUSION: The results of the present study showed that the functionality of HDL is impaired in ACS patients and that the impairment may be due to oxidative stress and an alteration of PON1 activities.


Assuntos
Síndrome Coronariana Aguda/metabolismo , Arildialquilfosfatase/metabolismo , Lipoproteínas HDL/metabolismo , Estresse Oxidativo , Síndrome Coronariana Aguda/sangue , Síndrome Coronariana Aguda/genética , Animais , Arildialquilfosfatase/genética , Índice de Massa Corporal , Linhagem Celular , Colesterol/metabolismo , Feminino , Humanos , Modelos Logísticos , Macrófagos/metabolismo , Masculino , Malondialdeído/sangue , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , Carbonilação Proteica , Vitamina E/sangue
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