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1.
Eur J Pharm Sci ; 194: 106692, 2024 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-38181870

RESUMO

In this work, five novel phosphonium salts derived from the Michael reaction were screened for their antiplatelet activity. Our findings revealed that compounds 2a, 2b, 2c, and 2d significantly inhibit platelet aggregation triggered by ADP or collagen (P < 0.001). Notably, compound 2c inhibited the arachidonic acid pathway (P < 0.001). Moreover, the selected compounds reduce CD62-P expression and inhibit GPIIb/IIIa activation. The interactions of the active compounds with their targets, ADP and collagen receptors, P2Y12 and GPVI respectively were investigated in silico using molecular docking studies. The results revealed a strong affinity of the active compounds for P2Y12 and GPVI. Additionally, cytotoxicity assays on platelets, erythrocytes, and human embryonic kidney HEK293 cells showed that compounds 2a, 2c and 2d were non-toxic even at high concentrations. In summary, our study shows that phosphonium salts can have strong antiplatelet power and suggests that compounds 2a, 2c and 2d could be promising antiplatelet agents for the management of cardiovascular diseases.


Assuntos
Inibidores da Agregação Plaquetária , Sais , Humanos , Simulação de Acoplamento Molecular , Células HEK293 , Inibidores da Agregação Plaquetária/farmacologia , Agregação Plaquetária , Plaquetas/metabolismo
2.
Foods ; 12(21)2023 Oct 24.
Artigo em Inglês | MEDLINE | ID: mdl-37959006

RESUMO

Glycosaminoglycans (GAGs) play a crucial role due to their significant biomedical functions. Chondroitin sulfate (CS) and dermatan sulfate (DS), the main representative family of GAGs, were extracted and purified from garfish (Belone belone) by-products, i.e., skin (GSB), bones (GCB), and heads (GHB), and their composition and anticoagulant activity were investigated. CS/DS were purified by ion-exchange chromatography with yields of 8.1% for heads, 3.7% for skin, and 1.4% for bones. Cellulose acetate electrophoresis was also explored for analyzing the extracted CS/DS. Interestingly, GHB, GSB, and GCB possessed sulfate contents of 21 ± 2%, 20 ± 1%, and 20 ± 1.5%, respectively. Physico-chemical analysis showed that there were no significant differences (p > 0.05) between the variances for sulfate, uronic acid, and total sugars in the GAGs extracted from the different parts of fish. Disaccharide analysis by SAX-HPLC showed that the GSB and GCB were predominately composed of ΔDi-4S [ΔUA-GalNAc 6S] (74.78% and 69.22%, respectively) and ΔDi-2,4S [ΔUA2S-GalNAc 4S] (10.92% and 6.55%, respectively). However, the GHB consisted of 25.55% ΔDi-6S [ΔUA-GalNAc 6S] and 6.28% ΔDi-2,6S [ΔUA2S-GalNAc 4S]. Moreover, classical anticoagulation tests were also used to measure their anticoagulant properties in vitro, which included the activated partial thromboplastin time, prothrombin time, and thrombin time. The CS/DS isolated from garfish by-products exhibited potent anticoagulant effects. The purified CS/DS showed exceptional anticoagulant properties according to this research and can be considered as a new agent with anticoagulant properties.

3.
BMC Complement Med Ther ; 22(1): 339, 2022 Dec 27.
Artigo em Inglês | MEDLINE | ID: mdl-36575459

RESUMO

BACKGROUND: Platelet aggregation and advanced glycation end products (AGEs) and oxidative stress are known as key factors for the development of cardiovascular diseases and diabetic complications. In this context, fruit and vegetable consumption, good sources of antioxidant compounds have been largely reported as an effective way of preventing human against these diseases. The current study focuses on the evaluation of antioxidant, antiplatelet and anti-glycation activities of pomegranate (Punica granatum L.) flowers (PF), leaves (PL), peel (PP) juice (PJ) and seeds oil (PSO). METHODS: Antioxidant activities was measured against ABTS radical and lipid peroxidation. Antiglycation activity was determined using the formation of AGE fluorescence intensity in the BSA/ribose system. Antiplatelet activity was measured in platelet rich plasma (PRP) against adenosine diphosphate (ADP), Collagen and arachidonic acid (AA). RESULTS: PF extract displayed the highest antioxidant activity against ABTS and lipid peroxidation with IC50 values of 0.7 mg/mL and 0.63 mg/mL respectively. For anti-glycation activity, PP, PF and PL inhibited moderately the pentosidine-like AGEs formation compared to positive controls with AGE-IC50 value of 0.4 mg/mL. PJ and PSO haven't any anti-AGE effect. All the extracts selectively inhibited platelet aggregation caused by one, two or three inducers in dose dependent manner. PF was the most potent inhibitor caused by all three inducers, with inhibitory effects ranging from 35.6 to 66.6%. PP and PJ exhibited antiplatelet effect against both ADP and collagen and PL and PSO only against AA. CONCLUSIONS: These results suggest that some pomegranate extracts exert potential in vitro anti-glycative and antiplatelet activities.


Assuntos
Antioxidantes , Punica granatum , Humanos , Antioxidantes/farmacologia , Extratos Vegetais/farmacologia , Frutas , Produtos Finais de Glicação Avançada , Colágeno , Difosfato de Adenosina
4.
Bioorg Chem ; 125: 105858, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35569189

RESUMO

This work focuses on the selection and the optimization of an efficient green-extraction method, used to recover a thymol-enriched extract from thyme (Thymus vulgaris L), as well as the evaluation of the inhibitory effect of this latter on the human platelet aggregation. Different innovative extraction techniques, namely bead milling extraction, ultrasound and microwave assisted extraction, were tested for their ability to recover a high added value extract from thyme. Among all tested eco-extraction techniques, microwave extraction (MAE) was the best method in term of its extraction yield (20.84% ± 0.51), thymol concentration (731.71 mg/g) and total phenolic (23.53 ± 1.83 mg (GAE)/g of extract) and flavonoid (6.22 ± 0.35 mg of QE/g of extract) contents. Moreover, thyme extract obtained by microwave assisted extraction (TMAE) showed the most active antioxidant effect comparing to the other tested extracts. Based on these results, TMAE was chosen to be evaluated for its antiplatelet effect. Thereby, arachidonic acid, collagen and ADP were used to induce the platelet aggregation on human platelet rich plasma taken from healthy controls and results revealed that TMAE strongly inhibited the induced platelet aggregation. Indeed, TMAE exhibited potent antiaggregant activity by inhibiting platelet activation, secretion and aggregation. Additionally, cytotoxicity assay on normal HEK-293 cells showed that TMAE has no cytotoxic effect even at high concentration (8 mg/ml) and can further be taken up to various biomedical applications mainly in the prevention of cardiovascular diseases.


Assuntos
Thymus (Planta) , Plaquetas , Células HEK293 , Humanos , Extratos Vegetais/farmacologia , Folhas de Planta , Timol/farmacologia
5.
Carbohydr Polym ; 253: 117283, 2021 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-33278949

RESUMO

The effect of extraction procedures on chemical composition, structural, antitumor and anticoagulant properties of the sulphated polysaccharide 'ulvan' from the green seaweed Ulva lactuca were investigated. The structural features of ulvans were carried out by FTIR and by one- and two- dimensional 1H and 13C NMR spectroscopic. The ulvans were mainly composed of rhamnose, xylose, and uronic acid. Chemical and spectroscopic analyses demonstrated that ulvans were constituted of (1→4)-ß-glucuronic acid, (1→3,4)-α-L-rhamnose-3-sulphate and (1→4)-α-xylose. The extraction procedures effect were observed in chemical structure, Mw and biological activities. Cytotoxic activity of enzymatic-chemical extract on cervical cancer cells (HeLa) (IC50 = 1000 µg/mL) was higher than on normal peripheral blood lymphocytes cells (PBL). Acid extracts promoted to reduce HeLa cells and to grow PBL cells. At high concentrations, acid extracts showed the highest APTT and TT clotting time. Antitumoral and anticoagulant activities of ulvans from Ulva lactuca promote their use as effective therapeutic agent.


Assuntos
Anticoagulantes/química , Anticoagulantes/farmacologia , Antineoplásicos/química , Antineoplásicos/farmacologia , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Polissacarídeos/química , Polissacarídeos/farmacologia , Ulva/química , Anticoagulantes/isolamento & purificação , Antineoplásicos/isolamento & purificação , Doadores de Sangue , Espectroscopia de Ressonância Magnética Nuclear de Carbono-13 , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células HeLa , Humanos , Leucócitos Mononucleares/efeitos dos fármacos , Tempo de Tromboplastina Parcial , Extratos Vegetais/isolamento & purificação , Polissacarídeos/isolamento & purificação , Alga Marinha/química , Espectroscopia de Infravermelho com Transformada de Fourier , Tempo de Trombina , Tunísia
6.
Int J Biol Macromol ; 164: 131-139, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-32673716

RESUMO

The present work deals with the extraction and purification of chondroitin sulfate/dermatan sulfate from skin (CSG) and bone (CBG) of corb (Sciaena umbra). Electrophoresis of these polymers in barium acetate buffer on cellulose acetate revealed two fractions similar to dermatan sulfate and chondroitin sulfate. The in vivo anticoagulant activity of both chondroitin sulfate/dermatan sulfate (CS/DS) were evaluated, at 25 and 75 mg kg-1 of body weight (b.w), using activated partial thromboplastin time (aPTT), prothrombine time (TT) and thrombin time (PT) tests. Results showed that aPTT of CSG and CBG at 75 mg kg-1 of b.w were prolonged by 1.59 and 1.48-fold respectively, compared with the control. Further, toxicity studies on liver performed by the catalytic activity of transaminases in plasma, oxidative stress markers and hepatic morphological changes demonstrated that CSG and CBG at both doses are not toxics. In summary, the higher activity and lower toxicity of both CS/DS, especially at 25 mg kg-1 of b.w, recommended these compounds as a better drug candidate.


Assuntos
Anticoagulantes/farmacologia , Sulfatos de Condroitina/farmacologia , Dermatan Sulfato/farmacologia , Peixes/metabolismo , Animais , Anticoagulantes/isolamento & purificação , Anticoagulantes/toxicidade , Testes de Coagulação Sanguínea , Osso e Ossos/química , Varredura Diferencial de Calorimetria , Sulfatos de Condroitina/isolamento & purificação , Sulfatos de Condroitina/toxicidade , Dermatan Sulfato/isolamento & purificação , Dermatan Sulfato/toxicidade , Avaliação Pré-Clínica de Medicamentos , Eletroforese em Acetato de Celulose , Feminino , Glicosaminoglicanos/isolamento & purificação , Fígado/efeitos dos fármacos , Testes de Função Hepática , Microscopia Eletrônica de Varredura , Estresse Oxidativo/efeitos dos fármacos , Ratos Wistar , Pele/química , Difração de Raios X
7.
Int J Biol Macromol ; 160: 593-601, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-32485252

RESUMO

The present study was undertaken to establish a distinct relationship between blue crab chitosan (Cs) acetylation degree (AD) and molecular weight (Mw) and its structural features, thermal properties and bioactivity. Therefore, chitosans with different AD were prepared and Cellulase was used to produce Cs derivatives with decreasing Mw. Results clearly display a decrease of the ordered structure of Cs, with the increase of AD and the decrease of Mw. Thermal stability/degradation screening disclose a greater thermal resistance for Cs with lower AD and higher Mw. The anti-adhesive potential of Cs was, additionally, studied, as function of AD and Mw. The effectiveness of Cs in preventing biofilm adhesion was strongly influenced by its AD and Mw, with the lowest inhibition values for higher AD and lower Mw. Interestingly, the effectiveness of Cs in disrupting pre-formed biofilms increased with decreasing Mw. Moreover, Cs derivatives were found to be advantageously efficient in prolonging human blood clotting times, based on data of activated partial thromboplastin time, Quick time and thrombin time assays, typically for the intrinsic coagulation pathway. Accordingly, depending on the predicted application of Cs, either in food, biomedical and pharmaceutical industries, AD and Mw are critical traits to be inevitably reflected on.


Assuntos
Adesivos/química , Adesivos/farmacologia , Coagulação Sanguínea/efeitos dos fármacos , Braquiúros/química , Quitosana/química , Quitosana/farmacologia , Acetilação/efeitos dos fármacos , Animais , Biofilmes/efeitos dos fármacos , Humanos , Peso Molecular , Trombina/metabolismo
8.
Tunis Med ; 97(2): 327-334, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31539091

RESUMO

AIMS: To determine region-specific reference ranges of lymphocyte T subsets in blood donors and to assess the influence of gender and age on lymphocyte T susbsets. METHODS: Blood samples from 143 blood donors were collected in the Blood Transfusion Center of Sfax. Lymphocyte T subsets were analyzed using a dual-platform method with a flow cytometer (percentages) and an automated hematology analyzer (white blood cells and lymphocytes). ANOVA and Student's tests were used for data analysis. RESULTS: Reference values were expressed as mean and 95% confidence intervals for T cells: CD3+: 1415 ± 348 cells/µL [1357-1473], CD3+/CD4+: 786 ± 220 cells/µL [732.31-811.7], CD3+/CD8+: 639 ± 258 cells/µL [596-862] and CD4+/CD8+ ratio was 1.46 ± 0.77 [1.36-1.62]. Gender and age influenced blood lymphocyte T subsets. CONCLUSION: Our study leads to the establishment of peripheral blood lymphocyte T subset reference ranges in blood donors in the region of Sfax. A study on a more diversified population, including more important number of individuals from various regions of Tunisia and including enumeration of other lymphocyte subsets (B cells and NK cells) is required.


Assuntos
Doadores de Sangue , Citometria de Fluxo/normas , Subpopulações de Linfócitos T/citologia , Adolescente , Adulto , Fatores Etários , Envelhecimento/sangue , Doadores de Sangue/estatística & dados numéricos , Complexo CD3/sangue , Relação CD4-CD8 , Feminino , Citometria de Fluxo/métodos , Humanos , Contagem de Linfócitos/normas , Contagem de Linfócitos/estatística & dados numéricos , Masculino , Pessoa de Meia-Idade , Valores de Referência , Linfócitos T/citologia , Linfócitos T/metabolismo , Tunísia/epidemiologia , Adulto Jovem
9.
Int J Biol Macromol ; 134: 405-412, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31071403

RESUMO

Chondroitin sulfate/dermatan sulfate (CS/DS) were isolated and purified for the first time from the bone of corb (Sciaena umbra) (CBG) and their chemical composition and anticoagulant activity were assessed. Infrared spectrum and agarose-gel electrophoresis for extracted CS/DS were also investigated. The results showed that the purified CS/DS obtained at a yield of 10% contains about 31.28% sulfate and an average molecular mass of 23.35 kDa. Disaccharide analysis indicated that CBG was composed of monosulfated disaccharides in positions 6 and 4 of the N-acetylgalactosamine (8.6% and 40.0%, respectively) and disulfated disaccharides in different percentages. The charge density was 1.4 and the ratio of 4:6 sulfated residues was equal to 4.64. Chondroitinase AC showed that the purified CS/DS contained mainly 74% CS and 26% DS. Moreover, the new CS/DS extracted from bone of corb showed a strong anticoagulant effect through activated partial thrombosis time (aPTT), thrombin time (TT) and prothrombin time (PT). In fact, CBG prolonged significantly (p < 0.05), aPTT and PT about 2.62 and 1.26 fold, respectively, greater than that of the negative control at a concentration of 1000 µg/mL. However, TT assay of CBG was prolonged 3.53 fold compared with the control at 100 µg/mL. The purified CS/DS displayed a promising anticoagulant potential, which may be used as a novel and soothing drug.


Assuntos
Anticoagulantes/química , Anticoagulantes/farmacologia , Osso e Ossos/química , Sulfatos de Condroitina/química , Sulfatos de Condroitina/farmacologia , Dermatan Sulfato/química , Dermatan Sulfato/farmacologia , Animais , Anticoagulantes/isolamento & purificação , Fracionamento Químico , Fenômenos Químicos , Sulfatos de Condroitina/isolamento & purificação , Dermatan Sulfato/isolamento & purificação , Peso Molecular , Umbridae
10.
Mar Drugs ; 17(4)2019 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-30934709

RESUMO

In this study, the antihypertensive activity of Purafect®-smooth hound viscera protein hydrolysate (VPH) and its peptide fraction with molecular weight (MW) below 1 kDa (VPH-I) was investigated. In addition, the lipase inhibitory activity, as well the anticoagulant potential, in vitro, were assessed. The antihypertensive effects of VPH and VPH-I were studied during 24 h (short-term effect) and 30 days (long-term effect) using high-salt (18% NaCl) and -fructose (10%) diet (HSFD)-induced hypertension. Data showed that, 4 h post-administration of VPH and VPH-I (200 mg/kg BW), the systolic blood pressure of rats was reduced by about 6 and 9 mmHg, respectively. These effects were similar to that obtained with Captopril (~9 mmHg at t = 4 h). On the other hand, exposing the rats to daily to HSFD, coupled to the administration of viscera peptides, was found to attenuate hypertension. In addition, the proteins' treatments were able to correct lipid and glycemic disorders, by reducing the total cholesterol and triglyceride contents and resorting to the plasma glucose level, compared to the HSFD group. Overall, the present findings demonstrated the preventive effect of VPH-peptides from hypertension complications, as a result of their biological properties.


Assuntos
Anti-Hipertensivos/farmacologia , Colesterol/metabolismo , Hipertensão/tratamento farmacológico , Hipertensão/prevenção & controle , Hidrolisados de Proteína/farmacologia , Animais , Glicemia/metabolismo , Pressão Sanguínea/efeitos dos fármacos , Dieta , Frutose/administração & dosagem , Metabolismo dos Lipídeos/efeitos dos fármacos , Masculino , Ratos , Ratos Wistar , Cloreto de Sódio/administração & dosagem
11.
Int J Biol Macromol ; 123: 335-342, 2019 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-30419328

RESUMO

A sulfated polysaccharide from Globularia alypum L. (GASP) was extracted with a yield of 14.2%. GASP is composed mostly of sulfate and total sugars (13.29% and 71.56%, respectively) with small amount of proteins and lipids. The chemical and structural characterization was studied by Infra-Red spectroscopic and gas chromatography-mass spectrometry (GC-MS). GASP composed of eight carbohydrates where galactose, glucose, and mannose are the major compounds (33.47%, 26.71% and 18.21%, respectively). The in vitro and in vivo anticoagulant activities in rats were tested using the standard coagulation assays activated partial thromboplastin time (aPTT), prothrombine time (TT) and thrombin time (PT) tests. Both doses of GASP (200 and 500 mg/kg b.w) displayed a significant in vitro (1.22 and 1.33-fold, 1.17 and 1.27-fold, and 1.21 and 1.26-fold, respectively) and in vivo (1.47 and 2.52-fold; 1.20 and 1.43-fold; 1.21 and 1.40-fold, respectively) compared with the control. Toxicity studies on liver performed by the catalytic activity of transaminases in plasma, oxidative stress markers and hepatic morphological changes indicated that GASP at both doses are not toxics. The important pharmacological and toxicological profile of GASP revealed that this compound may be used as a novel and effective drug.


Assuntos
Anticoagulantes/administração & dosagem , Coagulação Sanguínea/efeitos dos fármacos , Plantaginaceae/química , Polissacarídeos/administração & dosagem , Animais , Anticoagulantes/química , Anticoagulantes/isolamento & purificação , Testes de Coagulação Sanguínea , Cromatografia Gasosa , Humanos , Espectrometria de Massas , Tempo de Tromboplastina Parcial/métodos , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Ratos , Sulfatos/química , Tempo de Trombina/métodos
12.
RSC Adv ; 8(17): 9383-9393, 2018 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-35541829

RESUMO

This study was carried out to investigate the hypolipidemic, cardioprotective and anticoagulant properties of fish goby protein hydrolysates (GPHs) in rats fed a high fat and fructose diet (HFFD). Wistar rats were fed with HFFD for 2 months, coupled with the oral administration of GPHs and undigested goby protein (UGP). Compared with the standard diet, HFFD induced dyslipidemia and liver structure alterations, and increased pancreatic lipase activity. In addition, HFFD caused a significant increase in body weight. Interestingly, administration of UGP and GPHs to HFFD fed rats was efficacious in lowering serum total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-c) as well as hepatic TC and TG, and increased the serum high density lipoprotein cholesterol (HDL-c) content. Moreover, all treatments significantly decreased the atherogenic index and coagulant factor levels (thrombin and prothrombin). UGP and GPH administration also significantly decreased pancreatic lipase activity, which mitigates lipid accumulation. Similarly, UGP and its hydrolysates showed cardioprotective potential revealed by decreasing the risk of atherogenic and coronary artery disease and improving the liver architecture. The ex vivo plasma clotting test showed that GPHs exert a great therapeutic anticoagulant potential. The overall results demonstrated that GPH supplementation can counteract high-fat/fructose diet-induced obesity.

13.
RSC Adv ; 8(66): 37965-37975, 2018 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-35558578

RESUMO

Chondroitin sulfate/dermatan sulfate (CS/DS) was extracted from Atlantic bluefin tuna (Thunnus thynnus) skin (SGAT) and was purified and characterized. SGAT was characterized by acetate cellulose electrophoresis, FTIR spectroscopy, 13C NMR spectroscopy and SAX-HPLC. According to the results obtained for specific chondroitinases (ABC and AC) and the SAX-HPLC separation of generated unsaturated repeating disaccharides, the polymer was found to contain a disaccharide monosulfated in positions 6 and 4 of GalNAc and disulfated disaccharides in different percentages. These results were confirmed by 13C NMR experiments. The average molecular mass was 24.07 kDa, as determined by PAGE analysis. SGAT was evaluated for its in vitro anticoagulant activity via activated partial thromboplastin time, thrombin time and prothrombin time tests. The polymer showed strong inhibitory activity against angiotensin I-converting enzyme (IC50 = 0.25 mg mL-1). Overall, the results suggest that this newly extracted CS/DS can be useful for pharmacological applications.

14.
Exp Toxicol Pathol ; 69(1): 45-53, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27825753

RESUMO

The present study aimed to characterize and evaluate the in vitro and in vivo anticoagulant activity of sulfated glycosaminoglycans from the skins of smooth hound (SHSG) and grey triggerfish (GTSG). The analysis of SHSG and GTSG with acetate cellulose electrophoresis in Zn-acetate revealed the presence of hyaluronic acid (HA), chondroitin sulfate (CS) and dermatan sulfate (DS). Both glycosaminoglycans were evaluated for their in vitro anticoagulant activities using activated partial thromboplastin time (aPTT), thrombin time (TT) and prothrombine time (PT) tests. SHSG and GTSG and calciparin were tested as in vivo anticoagulants by subcutaneous (s.c) injection to adult female Wistar rats in a concentration of 75mg/kg of body weight. The administration of SHSG, GTSG and calciparin to rats induced a significant decrease of platelet rates compared to the control. The aPTT assay of SHSG and GTSG was prolonged 1.3 and 1.23-fold respectively compared with the control. Toxicity studies were performed to investigate whether or not SHSG and GTSG can cause pathological changes in the liver, proteins and DNA. The concentration and catalytic activity of liver oxidative stress markers and enzymes, respectively, as well as the observed hepatic morphological changes indicated that calciparin induced hepatic toxicity and oxidative damage in the liver. The higher activity and lower toxicity of SHSG and GTSG recommended these compounds as a better drug candidate than calciparin.


Assuntos
Anticoagulantes/farmacologia , Peixes , Glicosaminoglicanos/farmacologia , Animais , Anticoagulantes/química , Feminino , Glicosaminoglicanos/química , Ratos , Ratos Wistar , Pele/química
15.
Biomed Pharmacother ; 80: 322-330, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-27133072

RESUMO

Sulfated glycosaminoglycans (SGNL) were extracted for the first time from Norway lobster (Nephrops norvegicus) shell. The monosaccharide composition analysed by GC/MS revealed the presence of galacturonic acid, glucuronic acid, N-acetylgalactosamine and N-acetylglucosamine. The analysis of SGNL with acetate cellulose electrophoresis in Zn-acetate revealed the presence of heparan sulfate (HS) and dermatan sulfate (DS). SGNL were evaluated for their anticoagulant activities using activated partial thromboplastin time (aPTT), thrombin time (TT) and prothrombine time (PT) tests. After 21h incubation, HCT116 cell proliferation was inhibited (p<0.05) between 39.7 and 54.8% at 1.5-7.5mg/mL of SGNL. SGNL don't show hemolytic activity towards bovine erythrocytes and no cytotoxicity against the normal lymphocytes. The antiproliferative efficacy of these lobster glycosaminoglycans were probably related with the higher sulfate content. SGNL demonstrated promising antiproliferative and anticoagulant potential, which may be used as a novel, effective and promising antithrombotic agent.


Assuntos
Exoesqueleto/química , Anticoagulantes/farmacologia , Colo/patologia , Glicosaminoglicanos/isolamento & purificação , Glicosaminoglicanos/farmacologia , Nephropidae/química , Animais , Bovinos , Morte Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Eletroforese , Células HCT116 , Hemólise/efeitos dos fármacos , Humanos , Tempo de Tromboplastina Parcial , Tempo de Protrombina , Espectrofotometria Ultravioleta , Espectroscopia de Infravermelho com Transformada de Fourier
16.
Artigo em Inglês | MEDLINE | ID: mdl-26606109

RESUMO

Glycocalicin (GC) is a large extracellular proteolytic fragment of glycoprotein Ib, a membrane platelet component playing an essential role in the physiological processes of platelet adhesion and aggregation. GC contains the binding sites for thrombin and von Willebrand factor. GC circulates normally in vivo in significant concentrations and the plasma level of this protein reflects a complex function of factors including platelet count or platelet turnover. It can therefore serve as a good indicator for many diseases like hypoplastic thrombocytopenia and idiopathic thrombocytopenic purpura. For this reason, several purification assays have been previously described. In this work, we describe a novel analytical method for GC purification from human platelets based on preparative HPLC gel filtration followed by immuno-affinity chromatography on NHS activated column conjugated with specific antibody. Pure GC was obtained from tiny amount of starting material. Our protocol of GC purification is simple, fast and provides a pure end product.


Assuntos
Complexo Glicoproteico GPIb-IX de Plaquetas/isolamento & purificação , Cromatografia de Afinidade , Cromatografia Líquida de Alta Pressão , Humanos
17.
Chem Biol Interact ; 236: 1-6, 2015 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-25912554

RESUMO

The aim of the present study was to evaluate the acute and sub-chronic toxicity of lipopeptides mixture produced by Bacillus mojavensis A21 as well as their in vitro anticoagulant activity. A21 lipopeptides was given to mice at single dose from 75 mg to 1000 mg/kg body weight (bw). The median lethal dose (LD50) of A21 lipopeptides was about 550 mg/kg bw. Sub-chronic toxicity study for 28 days was done by daily oral administration of A21 lipopeptides at doses of 40 and 400 mg/kg bw in rats. Results showed that A21 lipopeptides did not cause any change in body weights and they did not produce any marked alterations in the hematological blood parameters including hematocrit concentration, hemoglobin level, white and red cells count. However, the platelets level decreased significantly compared to control value. Moreover, no significant differences in the serum biochemical characteristics were observed for rats treated by the lowest dose. In contrast, a little enhancement of alanine-aminotransferase (ALT) activity and decrease in total cholesterol were observed with the highest dose. A21 lipopeptides were also found to cause a prolongation of the thrombin time (TT), the prothrombin time (PT) and the activated partial thromboplastin time (APTT). Overall, A21 lipopeptides may be very promising compounds for therapeutic purposes.


Assuntos
Anticoagulantes/farmacologia , Bacillus/química , Lipopeptídeos/farmacologia , Lipopeptídeos/toxicidade , Administração Oral , Animais , Anticoagulantes/administração & dosagem , Anticoagulantes/toxicidade , Comportamento Animal/efeitos dos fármacos , Peso Corporal/efeitos dos fármacos , Lipopeptídeos/administração & dosagem , Masculino , Camundongos , Tamanho do Órgão/efeitos dos fármacos , Tempo de Tromboplastina Parcial , Tempo de Protrombina , Ratos Wistar , Tempo de Trombina , Testes de Toxicidade Aguda , Testes de Toxicidade Subcrônica
18.
Hematology ; 18(6): 365-9, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23601927

RESUMO

OBJECTIVES: Human platelet-specific alloantigens (HPA) are polymorphic epitopes which vary among ethnic groups. BACKGROUND: In Tunisia, HPA frequencies were determined in North and centre; however, the pattern of HPA in South Tunisian population is not been studied yet. The aim of this work was to determine allelic frequencies of HPA-1, -3, and -5 systems in south Tunisian population, in order to estimate the risk of anti-platelet allo-immunization and to create a register of HPA-typed blood donors. METHODS: Our study concerned 212 unrelated healthy, regular blood donors from southern Tunisia. Allelic polymorphisms of each system were determined using a polymerase chain reaction with sequence-specific primers. RESULTS: Genotype frequencies a/a, a/b, and b/b were, respectively, 0.670, 0.288, and 0.042 for HPA-1 system, 0.430, 0.462, and 0.108 for HPA-3 system, and 0.750, 0.241, and 0.009 for HPA-5 system. The allele frequencies were 0.814 and 0.186 for HPA-1a and -1b alleles; 0.660 and 0.340 for HPA-3a and -3b alleles and 0.870, and 0.130 for HPA-5a and -5b alleles. DISCUSSION: The reported frequencies are more similar to those of Caucasians than those of north Tunisian population.


Assuntos
Antígenos de Plaquetas Humanas/genética , Plaquetas/imunologia , Frequência do Gene , Antígenos de Plaquetas Humanas/sangue , Antígenos de Plaquetas Humanas/imunologia , Epitopos/genética , Epitopos/imunologia , Feminino , Genótipo , Humanos , Masculino , Reação em Cadeia da Polimerase , Polimorfismo Genético , Tunísia
19.
Cryobiology ; 65(3): 326-31, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23010483

RESUMO

We aimed in the first part of our work to study the effect of cryopreservation on the human sperm DNA integrity and the activation of caspase 3, the main apoptosis indicator. In the second part, we were interested in testing the effect of quercetin, as an antioxidant, in preventing sperm damage during the freeze-thawing process. Seventeen semen samples were obtained from 17 men recruited for infertility investigations. Liquefied sperm was cryopreserved using spermfreeze®. Nine of the used samples were divided into two aliquots; the first one was cryopreserved with spermfreeze only (control) and the second one was cryopreserved with spermfreeze supplemented with quercetin to a final concentration of 50 µM. Sperm motility and viability were assessed according to WHO criteria. We used TUNEL assay and the Oxy DNA assay to assess sperm DNA integrity. Activated caspase 3 levels were measured in spermatozoa using fluorescein-labeled inhibitor of caspase (FLICA). Cryopreservation led to a significant increase in sperm DNA fragmentation, DNA oxidation and caspase 3 activation (p<0.01). Supplementation of the cryopreservation medium with quercetrin induced a significant improvement in post thaw sperm parameters, compared to those of control, regarding sperm motility (p=0.007), viability (p=0.008) and DNA integrity (p=0.02); however, it had no effect on caspase 3 activation (p=0.3). We conclude that oxidative stress plays a major role in inducing sperm cryodamage but implication of apoptosis in this impairment requires further investigations. Quercetin could have protective effect during cryopreservation but further research is needed to confirm this effect.


Assuntos
Antioxidantes/farmacologia , Criopreservação/métodos , Quercetina/farmacologia , Preservação do Sêmen/métodos , Espermatozoides/citologia , Caspase 3/metabolismo , Sobrevivência Celular/efeitos dos fármacos , DNA/metabolismo , Fragmentação do DNA/efeitos dos fármacos , Humanos , Marcação In Situ das Extremidades Cortadas , Masculino , Estresse Oxidativo/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Espermatozoides/metabolismo
20.
Transfusion ; 52(1): 134-43, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22014098

RESUMO

BACKGROUND: The goal of selecting a healthy blood donor is to safeguard donors and reduce the risks of infections and immunologic complications for recipients. STUDY DESIGN AND METHODS: To evaluate the blood donor selection process, a survey was conducted in 28 blood transfusion centers located in 15 francophone African countries. Data collected included availability of blood products, risk factors for infection identified among blood donor candidates, the processing of the information collected before blood collection, the review process for the medical history of blood donor candidates, and deferral criteria for donor candidates. RESULTS: During the year 2009, participating transfusion centers identified 366,924 blood donor candidates. A mean of 13% (range, 0%-36%) of the donor candidates were excluded based solely on their medical status. The main risk factors for blood-borne infections were having multiple sex partners, sexual intercourse with occasional partners, and religious scarification. Most transfusion centers collected this information verbally instead of having a written questionnaire. The topics least addressed were the possible complications relating to the donation, religious scarifications, and history of sickle cell anemia and hemorrhage. Only three centers recorded the temperature of the blood donors. The deferral criteria least reported were sickle cell anemia, piercing, scarification, and tattoo. CONCLUSIONS: The medical selection process was not performed systemically and thoroughly enough, given the regional epidemiologic risks. It is essential to identify the risk factors specific to francophone African countries and modify the current medical history questionnaires to develop a more effective and relevant selection process.


Assuntos
Doadores de Sangue/estatística & dados numéricos , Transfusão de Sangue/estatística & dados numéricos , Seleção do Doador/métodos , Seleção do Doador/normas , Adulto , África , Bancos de Sangue/estatística & dados numéricos , Feminino , Humanos , Masculino
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