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BACKGROUND: Solvent-free microwave-assisted extraction (MAE) is a green extraction method capable of boosting the yield and quality profile of hemp essential oil when compared with other conventional extraction techniques. During this process, two by-products are produced, namely the aqueous residue containing bioactive phenolics and the residual deterpenated biomass, which can be used for further extraction and purification of phytocannabinoids. To date, the hemp industry has not utilized these products, although they can be valuable for the food, cosmetic, nutraceutical and pharmaceutical market. RESULTS: This study assessed and optimized the variables affecting MAE efficiency, namely microwave irradiation power, extraction time and added water, which were studied using a central composite design approach, and results were used to optimize the extraction process for recovering three valuable fractions: essential oil, polyphenols and phytocannabinoids. The products obtained using the optimized conditions were characterized in terms of yield, chemical profile and antioxidant potential. Moreover, the by-products obtained during the optimized run were further analyzed in terms of their biological activity using both enzymatic and non-enzymatic assays. The aqueous residue demonstrated a powerful α-glucosidase inhibition, a good activity in terms of superoxide radical scavenging activity, a modest efficacy in terms of inhibition of advanced glycation end products formation and no activity in terms of lipase inhibition. The residual deterpenated biomass did not possess significant biological activity. CONCLUSION: This work demonstrated valorization of industrial hemp essential oil and its by-products, obtained by a sustainable and eco-friendly extraction method, through an almost waste-free approach. Cannabinoids as well as other valuable bioactive compounds such as glycosidic flavones may be recovered from the residues of the essential oil extraction, representing interesting substances in the pharmaceutical, cosmetic and nutraceutical fields. © 2022 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
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Canabinoides , Cannabis , Flavonas , Óleos Voláteis , Antioxidantes/análise , Canabinoides/química , Cannabis/química , Produtos Finais de Glicação Avançada , Lipase , Micro-Ondas , Superóxidos , Água , alfa-GlucosidasesRESUMO
CONTEXT: Brassica incana Ten. (Brassicaceae) is an edible plant with very limited available information. Previous studies have demonstrated the polyphenolic profile and the antioxidant and cytotoxic properties of the leaf and flowering top hydroalcoholic extracts. OBJECTIVE: The volatile composition and the antidiabetic and anti-obesity potential of B. incana leaf and flowering top extracts have been investigated. MATERIAL AND METHODS: The volatile characterization of the extracts was attained by HS-SPME-GC/MS analysis. The antidiabetic and anti-obesity potential was investigated spectrophotometrically in vitro by the ability to modulate pancreatic lipase and α-glucosidase at different concentrations using orlistat and acarbose as reference drugs. The inhibition of advanced glycation end-products (AGEs) was measured with aminoguanidine as reference and the antioxidant activity with the xanthine/xanthine oxidase system and Trolox for comparative purposes. RESULTS: Several volatiles belonging to different chemical classes were identified, being sulphur compounds the most abundant in both leaf and flowering top extracts (56.33% and 64.40% of all volatiles). Although the leaf extract showed lower IC50 values in most of the assays (0.968 and 1.921 mg/mL for α-glucosidase; 0.192 and 0.262 mg/mL for AGEs; 0.022 and 0.038 mg/mL for superoxide scavenging), there were no statistically significant differences between both samples. These extracts showed a similar behaviour to Trolox in the xanthine oxidase assay (IC50 values of 0.022 mg/mL for leaf extract; 0.038 mg/mL for flowering top and 0.028 for Trolox). CONCLUSIONS: Leaves and flowering tops from B. incana can be used as sources of functional compounds that could act as antidiabetic and anti-obesogenic agents.
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Brassica , Hipoglicemiantes , Acarbose , Antioxidantes/química , Antioxidantes/farmacologia , Topos Floridos , Hipoglicemiantes/química , Hipoglicemiantes/farmacologia , Lipase , Orlistate , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Compostos de Enxofre , Superóxidos , Xantina Oxidase , Xantinas , alfa-GlucosidasesRESUMO
Chagas disease is initiated upon infection by Trypanosoma cruzi. Among the health consequences is a decline in heart function, and the pathophysiological mechanisms underlying this manifestation are not well understood. To explore the possible mechanisms, we employed IgY LC10 affinity chromatography in conjunction with ProteomeLab PF2D and two-dimensional gel electrophoresis to resolve the proteome signature of high and low abundance serum proteins in chagasic patients. MALDI-TOF MS/MS analysis yielded 80 and 14 differentially expressed proteins associated with cardiomyopathy of chagasic and other etiologies, respectively. The extent of oxidative stress-induced carbonyl modifications of the differentially expressed proteins (n = 26) was increased and coupled with a depression of antioxidant proteins. Functional annotation of the top networks developed by ingenuity pathway analysis of proteome database identified dysregulation of inflammation/acute phase response signaling and lipid metabolism relevant to production of prostaglandins and arachidonic acid in chagasic patients. Overlay of the major networks identified prothrombin and plasminogen at a nodal position with connectivity to proteome signature indicative of heart disease (i.e., thrombosis, angiogenesis, vasodilatation of blood vessels or the aorta, and increased permeability of blood vessel and endothelial tubes), and inflammatory responses (e.g., platelet aggregation, complement activation, and phagocyte activation and migration). The detection of cardiac proteins (myosin light chain 2 and myosin heavy chain 11) and increased levels of vinculin and plasminogen provided a comprehensive set of biomarkers of cardiac muscle injury and development of clinical Chagas disease in human patients. These results provide an impetus for biomarker validation in large cohorts of clinically characterized chagasic patients.
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Biomarcadores/sangue , Doença de Chagas/sangue , Proteoma/análise , Proteômica/métodos , Adolescente , Adulto , Idoso , Biomarcadores/metabolismo , Cardiomiopatia Chagásica/sangue , Cardiomiopatia Chagásica/metabolismo , Doença de Chagas/metabolismo , Eletroforese em Gel Bidimensional , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Carbonilação Proteica , Proteoma/classificação , Proteoma/metabolismo , Transdução de Sinais , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Adulto JovemRESUMO
Life expectancy has increased considerably in the last decades, clearing the way for preventive medicine. The ingestion of healthy foods or ingredients to improve health is gaining attention and edible flowers entail a promising source of bioactive compounds. The aim of this work was to study the anti-ageing and anti-obesity properties of an extract obtained from an edible flower Viola x wittrockiana though in vitro and in vivo methodologies with Caenorhabditis elegans as a model. The capacity to inhibit the enzymes α-glucosidase and lipase as well as to prevent advance glycation end-product (AGE) formation was tested in vitro. Caenorhabditis elegans was used as an obesity in vivo model to assess the effects of the extract on fat accumulation, development, progeny and health span. Viola flowers showed lower IC50 values in the α-glucosidase assay than the reference drug acarbose and exerted a higher inhibition of AGE formation than the reference substance aminoguanidine; the extract also showed pancreatic lipase inhibiting properties. Moreover, the extract lowered fat storage of C. elegans in a dose-dependent manner, up to 90.37% at the highest tested dose, and improved health span biomarkers such as lipofuscin accumulation and progeny availability. Our results demonstrate, for the first time, the anti-obesogenic and anti-ageing activity of Viola x wittrockiana flowers and their potential use as functional foods and nutraceuticals.
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Diabetes mellitus (DM) is a metabolic disease characterized by a high blood sugar level that can cause severe complications to the organism or even death when not treated. However, certain dietary habits and foods may have beneficial effects on this condition. A polyphenolic-rich extract (containing hyperoside, isoquercitrin, quercetin, ellagic acid, and vanillic acid) of Tageres erecta L. (T. erecta) was obtained from yellow and orange flowers using an ethanolic Soxhlet extraction. These extracts were screened for antidiabetic and anti-obesity properties using in vitro and in vivo procedures. The capacity to inhibit the enzymes lipase and α-glucosidase, as well as the inhibition of advance glycation end-products (AGEs) was tested in vitro. Caenorhabditis elegans (C. elegans) was used as an obesity in vivo model to assess extracts effects on fat accumulation using the wild-type strain N2 and a mutant with no N3 fatty acid desaturase activity BX24. Extracts from both cultivars (yellow and orange) T. erecta presented in vitro inhibitory activity against the enzymes lipase and α-glucosidase, showing lower IC50 values than acarbose (control). They also showed important activity in preventing AGEs formation. The polyphenol-rich matrices reduced the fat content of obese worms in the wild-type strain (N2) down to levels of untreated C. elegans, with no significant differences found between negative control (100% reduction) and both tested samples (p < 0.05). Meanwhile, the fat reduction was considerably lower in the BX24 mutants (fat-1(wa-9)), suggesting that N3 fatty acid desaturase activity could be partially involved in the T. erecta flower effect. Our findings suggested that polyphenols from T. erecta can be considered candidate bioactive compounds in the prevention and improvement of metabolic chronic diseases such as obesity and diabetes.
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Polifenóis , Tagetes , Animais , Polifenóis/farmacologia , Polifenóis/metabolismo , Caenorhabditis elegans/metabolismo , Hipoglicemiantes/farmacologia , Hipoglicemiantes/uso terapêutico , alfa-Glucosidases/metabolismo , alfa-Glucosidases/farmacologia , Extratos Vegetais/farmacologia , Extratos Vegetais/metabolismo , Flores , Obesidade/tratamento farmacológico , Lipase/metabolismo , Produtos Finais de Glicação Avançada/metabolismo , Produtos Finais de Glicação Avançada/farmacologia , Ácidos Graxos Dessaturases/metabolismo , Ácidos Graxos Dessaturases/farmacologiaRESUMO
Allium cepa L., commonly known as onion, is one of the most-consumed vegetables. The benefits of the intake of its bulb are well studied and are related to its high polyphenol content. The flowers of onions are also edible; however, there are no studies about their biological properties. Our aim was to determine the polyphenolic profile and assess the antioxidant and anti-obesity capacity of an ethanolic extract from fresh flowers of A. cepa. The phenolic constituents were identified through LC-DAD-ESI/MSn. For the anti-obesity potential, the inhibitory activity against digestive enzymes was measured. Several in vitro assays were carried out to determine the antioxidant capacity. A Caenorhabditis elegans model was used to evaluate the effect of the extract on stress resistance and fat accumulation. For the first time, kaempferol and isorhamnetin glucosides were identified in the flowers. The extract reduced fat accumulation in the nematode and had a high lipase and α- glucosidase inhibitory activity. Regarding the antioxidant activity, the extract increased the survival rate of C. elegans exposed to lethal oxidative stress. Moreover, the activities of superoxide dismutase and catalase were enhanced by the extract. Our results demonstrate, for the first time, the antioxidant and anti-obesity activity of onion flowers and their potential use as functional foods and nutraceuticals.
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Black mulberry, Morus nigra L. (family: Moraceae), is a healthy food and medicinal plant. Microwave hydrodiffusion and gravity (MHG) is one of the most innovative applications of solvent-free microwave extraction. The aim of this study was to optimize for the first time the MHG solvent-free extraction of polyphenols and sugars from M. nigra fruits. Optimization was carried out using a central composite design (CCD) with selected responses such as extraction yield, total polyphenol (TPC), flavonoid (TFC), anthocyanin (TAC), and sugar (TSC) contents, in addition to DPPH radical scavenging, and α-glucosidase (AGHi), lipase (Li), and xanthine oxidase (XOi) inhibition as tools to evaluate the best parameters for efficient and rapid extraction of black mulberry. The optimized extract was characterized in terms of the aforementioned parameters to validate the models, and was further analyzed for 36 individual polyphenols using HPLC-MS/MS. The optimized MHG extract was finally compared with traditional extracts, and demonstrated much better performance in terms of TPC, TAC, and Li, while the traditional extracts showed better XOi and AGHi. In conclusion, MHG is a valuable green technique for the production of non-degraded black mulberry polyphenol-rich extract and we suggest its larger use in the pharmaceutical and food industries.
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Helichrysum stoechas (L.) Moench (H. stoechas) is a medicinal plant traditionally used in the Iberian Peninsula to treat different disorders such as arterial hypertension. The aim of this study was to investigate the vascular effects of a polyphenolic methanolic extract of H. stoechas, which has high antioxidant activity, and its mechanism of action. Isometric myography studies were performed in an organ bath with rat aortic rings with intact endothelium. The H. stoechas extract produced vasorelaxation in the aortic rings that were precontracted by phenylephrine or KCl. L-NAME and Rp-8-Br-PET-cGMPS but not indomethacin or H-89; it also reduced the relaxant response evoked by H. stoechas extract on the phenylephrine-induced contractions. H. stoechas extract reduced the response to CaCl2 similar to verapamil and reduced the phenylephrine-induced contractions comparable with heparin. TRAM-34, apamin and glibenclamide reduced relaxation induced by the H. stoechas extract. The combination of L-NAME+TRAM-34+apamin almost completely inhibited the H. stoechas-induced effect. In conclusion, the relaxant effect of the H. stoechas extract is partially mediated by endothelium through the activation of the NO/PKG/cGMP pathway and the opening of Ca2+-activated K+ channels. Furthermore, the decrease in the cytosolic Ca2+ by the inhibition of Ca2+ influx through the L-type Ca2+ channels and by the reduction of Ca2+ release from the sarcoplasmic reticulum via the IP3 pathway is also involved.
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BACKGROUND: Birth defects (BDs) are a serious public health problem in Mexico. The objective of this paper was to identify the frequency of newborns (NBs) that are born small for gestational age with a birth defect. MATERIALS AND METHODS: A cross-sectional study of NBs from hospitals part of the Mexican Institute of Social Security in Chihuahua, Mexico, was conducted. NBs were classified according to the presence or absence of a BD and according to their weight percentile using regional standards of birth weight. RESULTS: NBs diagnosed with (n = 263) or without BDs (n = 64,626) were included in this study. A greater proportion of NBs small for gestational age were identified in cases involving BDs (20%), compared with NBs without BDs (9%). Moreover, the average decrease in birth weights of NBs with BDs associated with their digestive system was 210 grams (95% confidence interval [CI], -436/-12), with their genital organs 440 grams (95% CI, -730/-151), involving chromosomal abnormalities 230 grams (95% CI, -435/-26), or with their musculoskeletal system 289 grams (95% CI, -43/-147) according to the gestational age, sex, and condition of the NB (p < 0.05). CONCLUSIONS: A greater proportion of NBs with BDs were associated with a low birth rate for their gestational age. In addition, some BDs were found to be associated with an impaired birth weight more often than others This suggests that clinical decisions regarding NBs with congenital defects and a lower birth weight for their gestational age should be diagnosed and treated for additional nutritional considerations as needed. Birth Defects Research (Part A), 2011. © 2011 Wiley-Liss, Inc.
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Peso ao Nascer , Anormalidades Congênitas , Recém-Nascido Pequeno para a Idade Gestacional , Índice de Apgar , Estudos Transversais , Idade Gestacional , Humanos , Recém-Nascido , MéxicoRESUMO
INTRODUCTION: Introduction: the goal of this work was to evaluate the acceptance of various types of thickeners, specifically modified starch thickener and gum thickener, both with and without flavoring. Patients and methods: a randomized sample of 40 hospitalized patients with oropharyngeal dysphagia was recruited. The taste, smell, and appearance of each type of thickener were evaluated, as well as the volume of liquid ingested by the patients taking each type of thickener (modified starch thickener vs. gum thickener, both with and without flavoring). Results: the overall acceptance of gum thickener was significantly higher than that of modified starch thickener (7.45 (1.57) vs. 5.10 (2.43), respectively; p = 0.001). When a food flavor was added to the thickened water, the overall rating of the product was higher than when no flavor was added (7.70 (1.53) vs. 4.85 (2.16); p < 0.001). The difference between the daily volume of water consumed by the patients who received gum thickeners (928.33 (331.27) mL) and those who received starch thickeners (670.00 (288.35) mL) was statistically significant (p = 0.012). Patient consumption was also higher when flavoring was added as compared to when it was not (943.33 (302.45) mL) vs. (655.00 (304.60) mL; p = 0.005). Conclusion: the acceptances of the thickener and of water intake by patients with dysphagia were both significantly higher when using gum thickeners compared to starch thickeners, and when adding flavoring.
INTRODUCCIÓN: Introducción: el objetivo de este trabajo fue evaluar la aceptación de varios tipos de espesantes (almidón modificado frente a gomas) con y sin saborizante. Pacientes y métodos: se reclutaron 40 pacientes hospitalizados con disfagia orofaríngea. Se evaluaron el sabor, el olor y la apariencia de cada tipo de espesante, así como el volumen de líquido ingerido por los pacientes que tomaban cada tipo de espesante (espesante de almidón modificado vs. espesante de goma, ambos con o sin saborizante). Resultados: la aceptación general del espesante de goma fue significativamente mayor que la del almidón modificado (7,45 (1,57) vs. 5,10 (2,43); p = 0,001). Cuando se añadió un saborizante al agua espesada, la calificación general fue mejor (7,70 (1,53) frente a 4,85 (2,16); p < 0,001). La diferencia entre el volumen diario de agua consumida por los pacientes que recibieron espesantes de goma (928,33 (331,27) ml) y los que recibieron espesantes de almidón (670,00 (288,35) ml) fue estadísticamente significativa (p = 0,012). El consumo de líquido también fue mayor cuando se agregó el saborizante (943,33 (302,45) ml frente a 655,00 (304,60) ml; p = 0,005). Conclusión: la aceptación del espesante y la ingesta de agua por parte de los pacientes con disfagia fueron significativamente mayores cuando se utilizaron espesantes de goma, en comparación con los espesantes de almidón, y al agregar saborizantes.
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Transtornos de Deglutição/complicações , Aditivos Alimentares/administração & dosagem , Transtornos de Deglutição/dietoterapia , Humanos , Projetos Piloto , Amido/administração & dosagem , Amido/uso terapêutico , Viscosidade/efeitos dos fármacosRESUMO
The house fly, Musca domestica L. (Diptera: Muscidae), continues to be a primary pest of livestock facilities worldwide. This pest also has shown a propensity for pesticide resistance development when under high selection pressures. In this study the house fly strain FDm was created by a 20% contribution from each of five colonies collected from dairies in Florida with known imidacloprid resistance. The FDm strain was used to evaluate the level ofimidacloprid resistance after five selections near the LC70 value of each selected generation. Overall, the mean selection mortality was 72.7, with males being considerably more susceptible than females. The unselected (F0) FDm strain showed considerable susceptibility to imidacloprid after its creation, compared with the five parental strains. Between 9500 and 14,000 virgin house flies were used in each selection. After the fifth and final selection, a 331-fold increase in imidacloprid resistance at the LC70 was observed over the parental FDm strain. In parallel studies, the FDm strain showed increasing tolerance of the commercial imidacloprid product QuickBayt. These results suggest that livestock producers should use caution when choosing pesticides and consider rotating fly baits, as is encouraged with other pesticide treatment regimes on farms.
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Moscas Domésticas/efeitos dos fármacos , Imidazóis/farmacologia , Inseticidas/farmacologia , Nitrocompostos/farmacologia , Animais , Indústria de Laticínios/normas , Feminino , Florida , Masculino , Neonicotinoides , Controle de Pragas/métodosRESUMO
OBJECTIVE: Maternal infections may induce placental, amniotic and, potentially, fetal inflammatory responses. As cytokine responses may be mediated by oxidative stress, we determined whether the antioxidant N-acetyl-cysteine (NAC), can attenuate maternally induced amniotic and placental cytokine responses to maternal infection (modeled by lipopolysaccharide [LPS]). STUDY DESIGN: Gestation day 18 pregnant rats were (1) treated with LPS (100 microg/kg, body weight; intraperitoneally) alone; (2) pretreated with NAC (300 mg/kg body weight; intraperitoneally) 30 minutes before LPS; (3) posttreated with NAC 120 minutes after LPS; or (4) treated with NAC 30 minutes before and 120 minutes after LPS. Six hours after LPS administration, maternal serum and amniotic fluid interleukin-6 (IL-6) and IL-10 levels, and placental IL-6 messenger RNA levels were determined. RESULTS: LPS increased maternal serum IL-6 (50 +/- 25 to 3444 +/- 584 pg/mL) and IL-10 (40 +/- 20 to 958 +/- 339 pg/mL) and amniotic fluid IL-6 (59 +/- 25 to 891 +/- 128 pg/mL). Pretreatment and/or posttreatment with NAC attenuated IL-6 in the maternal serum and amniotic fluid and IL-10 in the amniotic fluid. LPS also induced placental IL-6 messenger RNA that was inhibited by treatment with NAC before and after LPS. CONCLUSION: NAC inhibition of inflammatory responses may protect the fetus from potential long-term sequelae.
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Acetilcisteína/farmacologia , Líquido Amniótico/metabolismo , Antioxidantes/farmacologia , Citocinas/antagonistas & inibidores , Mediadores da Inflamação/antagonistas & inibidores , Lipopolissacarídeos/farmacologia , Placenta/metabolismo , Animais , Feminino , Interleucina-10/sangue , Interleucina-6/sangue , Interleucina-6/genética , Interleucina-6/metabolismo , Gravidez , RNA Mensageiro/antagonistas & inibidores , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
BACKGROUND: We investigated the pathological and diagnostic role of selected markers of inflammation, oxidant/antioxidant status, and cellular injury in human Chagas disease. METHODS: Seropositive/chagasic subjects characterized as clinically-symptomatic or clinically-asymptomatic (n = 116), seronegative/cardiac subjects (n = 102), and seronegative/healthy subjects (n = 45) were analyzed for peripheral blood biomarkers. RESULTS: Seropositive/chagasic subjects exhibited an increase in sera or plasma levels of myeloperoxidase (MPO, 2.8-fold), advanced oxidation protein products (AOPP, 56%), nitrite (5.7-fold), lipid peroxides (LPO, 12-17-fold) and malondialdehyde (MDA, 4-6-fold); and a decline in superoxide dismutase (SOD, 52%) and glutathione (GSH, 75%) contents. Correlation analysis identified a significant (p<0.001) linear relationship between inflammatory markers (AOPP/nitrite: r = 0.877), inflammation and antioxidant/oxidant status (AOPP/glutathione peroxidase (GPX): r = 0.902, AOPP/GSH: r = 0.806, Nitrite/GPX: 0.773, Nitrite/LPO: 0.805, MDA/MPO: 0.718), and antioxidant/oxidant levels (GPX/MDA: r = 0.768) in chagasic subjects. Of these, MPO, LPO and nitrite biomarkers were highly specific and sensitive for distinguishing seropositive/chagasic subjects from seronegative/healthy controls (p<0.001, training and fitting AUC/ROC >0.95). The MPO (r = 0.664) and LPO (r = 0.841) levels were also correlated with clinical disease state in chagasic subjects (p<0.001). Seronegative/cardiac subjects exhibited up to 77% decline in SOD, 3-5-fold increase in LPO and glutamate pyruvate transaminase (GPT) levels, and statistically insignificant change in MPO, AOPP, MDA, GPX, GSH, and creatine kinase (CK) levels. CONCLUSIONS: The interlinked effects of innate immune responses and antioxidant/oxidant imbalance are major determinants of human Chagas disease. The MPO, LPO and nitrite are excellent biomarkers for diagnosing seropositive/chagasic subjects, and MPO and LPO levels have potential utility in identifying clinical severity of Chagas disease.
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Antioxidantes/análise , Biomarcadores/sangue , Doença de Chagas/imunologia , Doença de Chagas/patologia , Imunidade Inata , Oxidantes/sangue , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Chagas disease, caused by Trypanosoma cruzi, is endemic in Latin America and an emerging infectious disease in the US and Europe. We have shown TcG1, TcG2, and TcG4 antigens elicit protective immunity to T. cruzi in mice and dogs. Herein, we investigated antigenicity of the recombinant proteins in humans to determine their potential utility for the development of next generation diagnostics for screening of T. cruzi infection and Chagas disease. METHODS AND RESULTS: Sera samples from inhabitants of the endemic areas of Argentina-Bolivia and Mexico-Guatemala were analyzed in 1(st)-phase for anti-T. cruzi antibody response by traditional serology tests; and in 2(nd)-phase for antibody response to the recombinant antigens (individually or mixed) by an ELISA. We noted similar antibody response to candidate antigens in sera samples from inhabitants of Argentina and Mexico (n=175). The IgG antibodies to TcG1, TcG2, and TcG4 (individually) and TcG(mix) were present in 62-71%, 65-78% and 72-82%, and 89-93% of the subjects, respectively, identified to be seropositive by traditional serology. Recombinant TcG1- (93.6%), TcG2- (96%), TcG4- (94.6%) and TcG(mix)- (98%) based ELISA exhibited significantly higher specificity compared to that noted for T. cruzi trypomastigote-based ELISA (77.8%) in diagnosing T. cruzi-infection and avoiding cross-reactivity to Leishmania spp. No significant correlation was noted in the sera levels of antibody response and clinical severity of Chagas disease in seropositive subjects. CONCLUSIONS: Three candidate antigens were recognized by antibody response in chagasic patients from two distinct study sites and expressed in diverse strains of the circulating parasites. A multiplex ELISA detecting antibody response to three antigens was highly sensitive and specific in diagnosing T. cruzi infection in humans, suggesting that a diagnostic kit based on TcG1, TcG2 and TcG4 recombinant proteins will be useful in diverse situations.
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Antígenos de Protozoários/imunologia , Doença de Chagas/diagnóstico , Doença de Chagas/prevenção & controle , Vacinas Protozoárias/administração & dosagem , Vacinas Protozoárias/imunologia , Trypanosoma cruzi/imunologia , Adolescente , Adulto , Idoso , Anticorpos Antiprotozoários/sangue , América Central , Doença de Chagas/imunologia , Ensaio de Imunoadsorção Enzimática , Humanos , Imunoglobulina G/sangue , Pessoa de Meia-Idade , Sensibilidade e Especificidade , América do Sul , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/imunologia , Adulto JovemRESUMO
Trypanosoma cruzi elicits reactive oxygen species (ROS) of inflammatory and mitochondrial origin in infected hosts. In this study, we examined ROS-induced oxidative modifications in the heart and determined whether the resultant oxidized cardiac proteins are targets of immune response and of pathological significance in Chagas disease. Heart biopsies from chagasic mice, rats and human patients exhibited, when compared to those from normal controls, a substantial increase in protein 4-hydroxynonenal (4-HNE), malondialdehyde (MDA), carbonyl, and 3-nitrotyrosine (3-NT) adducts. To evaluate whether oxidized proteins gain antigenic properties, heart homogenates or isolated cardiomyocytes were oxidized in vitro and one- or two-dimensional gel electrophoresis (2D-GE)/Western blotting (WB) was performed to investigate the proteomic oxidative changes and recognition of oxidized proteins by sera antibodies in chagasic rodents (mice, rats) and human patients. Human cardiomyocytes exhibited LD(50) sensitivity to 30 µM 4-HNE and 100 µM H(2)O(2) at 6 h and 12 h, respectively. In vitro oxidation with 4-HNE or H(2)O(2) resulted in a substantial increase in 4-HNE- and carbonyl-modified proteins that correlated with increased recognition of cardiac (cardiomyocytes) proteins by sera antibodies of chagasic rodents and human patients. 2D-GE/Western blotting followed by MALDI-TOF-MS/MS analysis to identify cardiac proteins that were oxidized and recognized by human chagasic sera yielded 82 unique proteins. We validated the 2D-GE results by enzyme-linked immunosorbent assay (ELISA) and WB and demonstrated that oxidation of recombinant titin enhanced its immunogenicity and recognition by sera antibodies from chagasic hosts (rats and humans). Treatment of infected rats with phenyl-α-tert-butyl nitrone (PBN, antioxidant) resulted in normalized immune detection of cardiac proteins associated with control of cardiac pathology and preservation of heart contractile function in chagasic rats. We conclude that ROS-induced, cardiac-oxidized antigens are targets of immune recognition by antibodies and molecular determinants for pathogenesis during Chagas disease.
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Autoanticorpos/imunologia , Doença de Chagas/etiologia , Doença de Chagas/imunologia , Miocárdio/imunologia , Espécies Reativas de Oxigênio/imunologia , Animais , Afinidade de Anticorpos , Antígenos/imunologia , Antígenos/metabolismo , Cardiomiopatia Chagásica/etiologia , Cardiomiopatia Chagásica/imunologia , Cardiomiopatia Chagásica/metabolismo , Cardiomiopatia Chagásica/patologia , Doença de Chagas/metabolismo , Doença de Chagas/parasitologia , Ensaios Clínicos como Assunto , Coração/parasitologia , Humanos , Camundongos , Camundongos Endogâmicos C3H , Miocárdio/metabolismo , Miócitos Cardíacos/imunologia , Miócitos Cardíacos/metabolismo , Miócitos Cardíacos/parasitologia , Oxirredução , Ligação Proteica , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Trypanosoma cruzi/imunologiaRESUMO
INTRODUCTION: In El Salvador, end-stage renal disease is the leading cause of hospital deaths in adults, the second cause of death in men and the fifth leading cause of death in adults of both sexes in the general population. OBJECTIVE: Identify risk factors for chronic kidney disease and urinary markers of renal and vascular damage, measure kidney function and characterize prevalence of chronic kidney disease in persons aged ≥18 years in the Bajo Lempa region of El Salvador. METHODS: A cross-sectional analytical epidemiological study was carried out using active screening for chronic kidney disease and associated risk factors in individuals aged ≥18 years in the Bajo Lempa Region, a rural, coastal area in El Salvador. Door-to-door visits and clinical examinations were conducted. Epidemiological and clinical data were collected including: family and personal clinical history of disease; biological, behavioral, social and environmental risk factors; physical measurements; urinalysis for markers of renal and vascular damage; and blood tests (serum creatinine, serum glucose, lipid profile). Glomerular filtration rate was calculated using the Modification of Diet in Renal Disease formula. Chronic kidney disease case confirmation was done three months later. Multiple logistic regression was used for data analysis. RESULTS: A total of 375 families and 775 individuals (343 men, 432 women) were studied-88.3% of the total resident population in the region. Elevated prevalence of risk factors was observed: diabetes mellitus, 10.3%; hypertension,16.9%; family history of chronic kidney disease, 21.6%; dyslipidemias, 63.1%; overweight, 34%; obesity, 22.4%; metabolic syndrome, 28.8%; use of non-steroidal anti-inflammatory drugs, 74.8%; infectious diseases, 86.9%; agricultural occupation, 40.6% (80.6% in men); and contact with agrochemicals, 50.3% (82.5% in men). Prevalence renal damage markers was 15.8% (greater in men): microalbuminuria 6.3%; proteinuria 5.7%; hematuria 3.5%; proteinuria-hematuria 0.3%. Proteinuria of <1 g/L predominated. Prevalence of chronic kidney disease was 17.9% (25.7% in men; 11.8% in women). Distribution by stages: stage 1, 4.6%; stage 2, 3.5%; stage 3, 6.2%; stage 4, 3.0%; stage 5, 0.6%. In patients with chronic kidney disease, most common was non-diabetic chronic kidney disease (86.3%), followed by chronic kidney disease associated with neither diabetes nor hypertension (54.7%). Prevalence of chronic renal failure was 9.8% (17% in men; 4.1% in women). Multiple logistic regression showed significant association with increasing age, male sex, hypertension and family history of chronic kidney disease. CONCLUSIONS: Elevated prevalence of chronic kidney disease, chronic renal failure and risk factors was found, compared to international reports. Most common was chronic kidney disease of unknown cause, associated with neither diabetes nor hypertension. Associations were found with age, male sex, hypertension and family history of chronic kidney disease, with decline in kidney function beginning at early ages. Male farmers have a dual burden of non traditional (occupational, toxic environmental) and traditional (vascular) risk factors that could act in synergy, contributing to kidney damage.
INTRODUCCIÓN En El Salvador, la enfermedad renal crónica terminal es la causa principal de muerte hospitalaria en adultos, la segunda causa de muerte en los hombres y la quinta causa principal de muerte entre adultos de ambos sexos en la población general. OBJETIVO Identificar los factores de riesgo de la enfermedad renal crónica y los marcadores de daño renovascular en orina, medir la función renal y caracterizar la prevalencia de enfermedad renal crónica en personas X18 años de edad en la región del Bajo Lempa en El Salvador. METODOS Se realizó un estudio epidemiológico transversal y analítico de la enfermedad renal crónica y los factores de riesgo asociados en individuos con edades X18 años por medio de pesquisa activa en la Región del Bajo Lempa, una zona costera rural en El Salvador. Se efectuaron visitas casa por casa y consultas médicas. Se recolectaron datos epidemiológicos y clínicos que incluían: historia clínica personal y familiar para la enfermedad; factores de riesgo biológicos, de conducta, sociales y ambientales; mediciones físicas; análisis de orina buscando marcadores de daño renovascular y exámenes de sangre (creatinina y glucosa en suero, lipidograma). La tasa de filtración glomerular se calculó usando la fórmula de MDRD (sigla en inglés de Modificación de la Dieta en la Enfermedad Renal). La confirmación de casos de enfermedad renal crónica se realizó en un período de tres meses. Se utilizó la regresión logística múltiple para analizar los datos. RESULTADOS Se estudiaron un total de 375 familias y 775 individuos (343 hombres, 432 mujeres), el 88,3% del total de la población residente en la región. Se observó una elevada prevalencia de factores de riesgo: diabetes mellitus en 10,3%; hipertensión en 16,9%; antecedentes familiares de enfermedad renal crónica en 21,6%; dislipidemias en 63,1%; sobrepeso en 34%; obesidad en 22,4%; síndrome metabólico en 28,8%; uso de medicamentos anti-inZ amatorios no esteroideos en 74,8%, enfermedades infecciosas en 86,9%, 40,6 % eran agricultores (80,6% de los hombres) y 50,3% tenían contacto con agroquímicos (82,5% de los hombres). La prevalencia de marcadores de daño renal fue de 15,8% (mayor en los hombres): microalbuminuria de 6,3%; proteinuria de 5,7%; hematuria de 3,5%; proteinuria-hematuria de 0,3%. Predominó la proteinuria <1 g/L. La prevalencia de enfermedad renal crónica fue de 17,9% (25,7 en los hombres; 11,8% en las mujeres). La distribución por estadios fue la siguiente: estadio 1, un 4,6%; estadio 2, un 3,5%' estadio 3, un 6,2%; estadio 4, un 3,0% y en estadio 5, un 0,6%. En pacientes con enfermedad renal crónica, la más común fue la enfermedad renal crónica no diabética (86,3%), seguida de la enfermedad renal crónica no asociada a diabetes ni a hipertensión (54,7%). La prevalencia de la insuficiencia renal crónica fue de 9,8% (17% en los hombres y 4,1% en mujeres). La regresión logística múltiple mostro una asociación significativa con el aumento de la edad, sexo masculino, hipertensión y antecedentes familiares de enfermedad renal crónica. CONCLUSIONES Se halló una elevada prevalencia de enfermedad renal crónica, insuficiencia renal crónica y factores de riesgo en comparación con lo reportado internacionalmente. La más frecuente fue la enfermedad renal crónica de causa desconocida, no asociada con diabetes ni hipertensión. Se encontraron asociaciones con la edad, sexo masculino, hipertensión e historia familiar de enfermedad renal crónica, con una disminución de la función renal que comienza a edades tempranas. Los agricultores hombres tienen la doble carga de riesgos no tradicionales (ocupacionales, toxico-ambientales) y tradicionales (vasculares) que podrían actuar de manera sinérgica, contribuyendo al daño renal. PALABRAS CLAVE Enfermedad renal crónica/epidemiologia, factores de riesgo, prevalencia, salud ocupacional, salud ambiental, plaguicidas, agroquímicos, El Salvador
Assuntos
Falência Renal Crônica/epidemiologia , Adulto , Biomarcadores/urina , Causas de Morte , Estudos Transversais , El Salvador/epidemiologia , Feminino , Humanos , Modelos Logísticos , Masculino , Programas de Rastreamento , Prevalência , Fatores de Risco , População RuralRESUMO
BACKGROUND: The housefly, Musca domestica L., continues to be a major pest of confined livestock operations. Houseflies have developed resistance to most chemical classes, and new chemistries for use in animal agriculture are increasingly slow to emerge. Five adult housefly strains from four Florida dairy farms were evaluated for resistance to four insecticides (beta-cyfluthrin, permethrin, imidacloprid and nithiazine). RESULTS: Significant levels of tolerance were found in most field strains to all insecticides, and in some cases substantial resistance was apparent (as deduced from comparison with prior published results). At the LC(90) level, greater than 20-fold resistance was found in two of the fly strains for permethrin and one fly strain for imidacloprid. Beta-cyfluthrin LC(90) resistance ratios exceeded tenfold resistance in three fly strains. The relatively underutilized insecticide nithiazine had the lowest resistance ratios; however, fourfold LC(90) resistance was observed in one southern Florida fly strain. Farm insecticide use and its impact on resistance selection in Florida housefly populations are discussed. CONCLUSION: Housefly resistance to pyrethroids is widespread in Florida. Imidacloprid resistance is emerging, and tolerance was observed to both imidacloprid and nithiazine. If these insecticides are to retain efficacy, producer use must be restrained.
Assuntos
Indústria de Laticínios , Inseticidas/farmacologia , Muscidae/efeitos dos fármacos , Piretrinas/farmacologia , Animais , Feminino , Florida , Imidazóis/farmacologia , Resistência a Inseticidas , Muscidae/fisiologia , Neonicotinoides , Nitrocompostos/farmacologia , Permetrina/farmacologiaRESUMO
OBJECTIVE: To identify the number, type, and appropriateness of abbreviations used to write nurse clinical charts (CC) and nursing discharge reports (NDR) in an inpatient unit. METHOD: We performed a descriptive cross-sectional study in an inpatient unit in the Hospital Clínico San Carlos in Madrid in November 2004. A sample of the CCs and NDRs of patients admitted to the unit during the period studied was analyzed. RESULTS: We analyzed 75 episodes, in which 405 abbreviations/acronyms and 9 different symbols were identified among a total of 1,201. Most of the abbreviations (85.7%; 95% confidence interval [CI] 81.9-88.8) were not included in the "Dictionary of medical abbreviations and other acronyms". For more than half of the abbreviated terms (58.5%; 95% CI: 53.7-63.2) several different abbreviations were used; the median number of abbreviations for the same term was 5 (P25: 2; P75: 4). CONCLUSIONS: A high percentage of the abbreviations used were nonstandard and their meaning had to be deduced from the context. Although only a small percentage of abbreviations were found in the NDRs, abbreviations should not be used in this type of document.
Assuntos
Abreviaturas como Assunto , Registros de Enfermagem/normas , Estudos Transversais , Estudos Retrospectivos , EspanhaRESUMO
Perinatal infections are a risk factor for fetal neurological pathologies, including cerebral palsy and schizophrenia. Cytokines that are produced as part of the inflammatory response are proposed to partially mediate the neurological injury. This study investigated the effects of intraperitoneal injections of lipopolysaccharide (LPS) to pregnant rats on the production of cytokines and stress markers in the fetal environment. Gestation day 18 pregnant rats were treated with LPS (100 microg/kg body wt i.p.), and maternal serum, amniotic fluid, placenta, chorioamnion, and fetal brain were harvested at 1, 6, 12, and 24 h posttreatment to assay for LPS-induced changes in cytokine protein (ELISA) and mRNA (real-time RT-PCR) levels. We observed induction of proinflammatory cytokines interleukin (IL)-1 beta, IL-6, and tumor necrosis factor-alpha (TNF-alpha) as well as the anti-inflammatory cytokine IL-10 in the maternal serum within 6 h of LPS exposure. Similarly, proinflammatory cytokines were induced in the amniotic fluid in response to LPS; however, no significant induction of IL-10 was observed in the amniotic fluid. LPS-induced mRNA changes included upregulation of the stress-related peptide corticotropin-releasing factor in the fetal whole brain, TNF-alpha, IL-6, and IL-10 in the chorioamnion, and TNF-alpha, IL-1 beta, and IL-6 in the placenta. These findings suggest that maternal infections may lead to an unbalanced inflammatory reaction in the fetal environment that activates the fetal stress axis.
Assuntos
Líquido Amniótico/metabolismo , Química Encefálica/efeitos dos fármacos , Hormônio Liberador da Corticotropina/metabolismo , Citocinas/biossíntese , Lipopolissacarídeos/farmacologia , Líquido Amniótico/efeitos dos fármacos , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/embriologia , Ensaio de Imunoadsorção Enzimática , Feminino , Placenta/efeitos dos fármacos , Placenta/metabolismo , Gravidez , Pró-Opiomelanocortina/metabolismo , RNA Mensageiro/biossíntese , RNA Mensageiro/isolamento & purificação , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Estresse Fisiológico/metabolismoRESUMO
Introducción: La disfunción endotelial es un marcador temprano de la enfermedad arterioesclerótica, y su detección nos permite determinar cuáles pacientes son potenciales sujetos de tratamiento para revertir la progresión del padecimiento vascular. Dado que el paciente diabético es un paciente con alta prevalencia de complicaciones arterioscleróticas, la detección de éste fenómeno en el mismo, es de capital importacia. Métodos y Resultados: Mediante Doppler braquial en estado basal, isquemia y post-nitroglicerina, se determinó la capacidad vascular de aumentar el flujo sanguíneo en 10 pacientes sanos y 20 pacientes diabéticos, con el objetivo de correlacionarlos con su HbA1c y perfil lipídico. La vasodilatación mediada por endotelio (VMF) y la vasodilatación mediada por nitroglicerina (VMN) expresadas en porcentaje de cambio del diámetro vascular, se encontraron reducidas en forma estadísticamente significativa en el grupo de diabéticos, (5.57 por ciento vs 9.97 por ciento para el grupo control p = 0.004 y 21.49 por ciento vs 12.64 por ciento para el grupo control p = 0.004 respectivamente) éstas alteraciones correlacionaron con los años de evolución de la enfermedad y los niveles de colesterol total, LDL y triglicéridos en éstos pacientes. Conclusión: El estudio doppler braquial es un instrumento útil para determinar mediante la disfunción endotelial, las fases tempranas de la arterioesclerosis en pacientes de alto riesgo para este fenómeno podrían ser intervenidos en aras de mejorar el pronóstico de estos pacientes. Palabras claves: Disfunción endotelial, Diabetes Mellitus.