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The Spanish Familial Pancreatic Cancer Registry (PANGENFAM) was established in 2009 and aims to characterize the genotype and phenotype of familial pancreatic cancer (FPC). Furthermore, an early detection screening program for pancreatic ductal adenocarcinoma (PDAC) is provided to healthy high-risk individuals from FPC and hereditary pancreatic cancer families (first-degree relatives). This article describes our experience over the last 10 years in high-risk screening. Hereditary and familial pancreatic cancer families were identified through the oncology and gastroenterology units. High-risk individuals underwent annual screening with endoscopic ultrasound (EUS) and magnetic resonance (MRI) from age 40 or 10 years younger than the youngest affected family member. Results: PANGENFAM has enrolled 290 individuals from 143 families, including 52 PDAC cases and 238 high-risk individuals. All high-risk individuals eligible for screening were offered to enter the surveillance program, with 143 currently participating. Pancreatic abnormalities were detected in 94 individuals (median age 53 years (29-83), with common findings including cystic lesions and inhomogeneous parenchyma. Imaging test concordance was 66%. Surgical intervention was performed in 4 high-risk individuals following highly suspicious lesions detected by imaging. PANGENFAM is a valuable resource for science innovation, such as biobanking, with clinical and imaging data available for analysis. For high-risk families, it may offer a potential for early diagnosis. Collaboration with other national and international registries is needed to increase our understanding of the disease biology and to standardize criteria for inclusion and follow-up, optimizing cost-effectiveness and efficacy.
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Carcinoma Ductal Pancreático , Neoplasias Pancreáticas , Sistema de Registros , Humanos , Neoplasias Pancreáticas/genética , Pessoa de Meia-Idade , Espanha , Feminino , Masculino , Adulto , Idoso , Seguimentos , Carcinoma Ductal Pancreático/genética , Carcinoma Ductal Pancreático/diagnóstico por imagem , Carcinoma Ductal Pancreático/diagnóstico , Idoso de 80 Anos ou mais , Predisposição Genética para Doença , Detecção Precoce de Câncer/métodos , Imageamento por Ressonância Magnética , Endossonografia , Fatores de Risco , CarcinomaRESUMO
Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of cancer-related death in the Western world. The number of diagnosed cases and the mortality rate are almost equal as the majority of patients present with advanced disease at diagnosis. Between 4 and 10% of pancreatic cancer cases have an apparent hereditary background, known as hereditary pancreatic cancer (HPC) and familial pancreatic cancer (FPC), when the genetic basis is unknown. Surveillance of high-risk individuals (HRI) from these families by imaging aims to detect PDAC at an early stage to improve prognosis. However, the genetic basis is unknown in the majority of HRIs, with only around 10-13% of families carrying known pathogenic germline mutations. The aim of this study was to assess an individual's genetic cancer risk based on sex and personal and family history of cancer. The Best Linear Unbiased Prediction (BLUP) methodology was used to estimate an individual's predicted risk of developing cancer during their lifetime. The model uses different demographic factors in order to estimate heritability. A reliable estimation of heritability for pancreatic cancer of 0.27 on the liability scale, and 0.07 at the observed data scale as obtained, which is different from zero, indicating a polygenic inheritance pattern of PDAC. BLUP was able to correctly discriminate PDAC cases from healthy individuals and those with other cancer types. Thus, providing an additional tool to assess PDAC risk HRI with an assumed genetic predisposition in the absence of known pathogenic germline mutations.
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Carcinoma Ductal Pancreático , Predisposição Genética para Doença , Mutação em Linhagem Germinativa , Neoplasias Pancreáticas , Humanos , Carcinoma Ductal Pancreático/genética , Neoplasias Pancreáticas/genética , Feminino , Masculino , Pessoa de Meia-Idade , Adulto , Idoso , Medição de Risco/métodos , CarcinomaRESUMO
BACKGROUNDPassive immunotherapy with convalescent plasma (CP) is a potential treatment for COVID-19. Evidence from controlled clinical trials is inconclusive.METHODSWe conducted a randomized, open-label, controlled clinical trial at 27 hospitals in Spain. Patients had to be admitted for COVID-19 pneumonia within 7 days from symptom onset and not on mechanical ventilation or high-flow oxygen devices. Patients were randomized 1:1 to treatment with CP in addition to standard of care (SOC) or to the control arm receiving only SOC. The primary endpoint was the proportion of patients in categories 5 (noninvasive ventilation or high-flow oxygen), 6 (invasive mechanical ventilation or extracorporeal membrane oxygenation [ECMO]), or 7 (death) at 14 days. Primary analysis was performed in the intention-to-treat population.RESULTSBetween April 4, 2020, and February 5, 2021, 350 patients were randomly assigned to either CP (n = 179) or SOC (n = 171). At 14 days, proportion of patients in categories 5, 6, or 7 was 11.7% in the CP group versus 16.4% in the control group (P = 0.205). The difference was greater at 28 days, with 8.4% of patients in categories 5-7 in the CP group versus 17.0% in the control group (P = 0.021). The difference in overall survival did not reach statistical significance (HR 0.46, 95% CI 0.19-1.14, log-rank P = 0.087).CONCLUSIONCP showed a significant benefit in preventing progression to noninvasive ventilation or high-flow oxygen, invasive mechanical ventilation or ECMO, or death at 28 days. The effect on the predefined primary endpoint at 14 days and the effect on overall survival were not statistically significant.TRIAL REGISTRATIONClinicaltrials.gov, NCT04345523.FUNDINGGovernment of Spain, Instituto de Salud Carlos III.
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COVID-19/terapia , SARS-CoV-2 , Idoso , COVID-19/mortalidade , COVID-19/fisiopatologia , Terapia Combinada , Progressão da Doença , Feminino , Hospitalização , Humanos , Imunização Passiva/efeitos adversos , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Razão de Chances , Pandemias , Espanha/epidemiologia , Resultado do Tratamento , Soroterapia para COVID-19RESUMO
INTRODUCTION: Anemia is a public health problem worldwide and is most prevalent in preschool children, for whom it is the most frequent cause of nutritional deficits. In turn, iron deficiency is the main cause of anemia, affecting 43% of children globally. Previous studies in Cuba show rates of iron deficiency in preschool children between 38.6% and 57.6%, higher in infants (71.2% to 81.1%). WHO recommends using serum ferritin as an indicator of iron deficiency accompanied by acute (C-reactive protein) and chronic (a1-acid glycoprotein) inflammation biomarkers. OBJECTIVE: Assess how inflammation affects measuring and reporting of iron-deficiency anemia rates in Cuban preschool children. METHODS: Data were obtained from serum samples contained in the National Anemia and Iron Deficiency Survey, and included presumably healthy preschool Cuban children (aged 6-59 months). Serum samples were collected from 1375 children from randomly selected provinces in 4 regions of the country from 2014 through 2018. We examined the association between ferritin and two inflammatory biomarkers: C-reactive protein and a1-acid glycoprotein. Individual inflammation-adjusted ferritin concentrations were calculated using four approaches: 1) a higher ferritin cut-off point (⟨30 g/L); 2) exclusion of subjects showing inflammation (C-reactive protein ⟩5 mg/L or a1-acid glycoprotein ⟩1 g/L); 3) mathematical correction factor based on C-reactive protein or a1-acid glycoprotein; and 4) correction by regression with the method proposed by the Biomarkers Reflecting Inflammation and Nutritional Determinants of Anemia Group. We estimated confidence intervals of differences between unadjusted prevalence and prevalence adjusted for inflammation by each method. RESULTS: The proportion of children with inflammation according to C-reactive protein concentrations >5 mg/L was lower (11.1%, 153/1375) than the proportion measured according to the concentrations of a1-acid glycoprotein, at >1 g/L (30.8%, 424/1375). The percentage of children with high concentrations of at least one of the aforementioned biomarkers was 32.7% (450/1375). Thus, each correction method increased the observed prevalence of iron deficiency compared to unadjusted estimates (23%, 316/1375). This increase was more pronounced when using the internal regression correction method (based only on C-reactive protein) or the method based on a higher cut-off point. Adjustment using all four methods changed estimated iron deficiency prevalence, increasing it from 0.1% to 8.8%, compared to unadjusted values. CONCLUSIONS: One-third of preschool children had biomarkers indicating elevated inflammation levels. Without adjusting for inflammation, iron deficiency prevalence was underestimated. The significant disparity between unadjusted and inflammation-adjusted ferritin when using some approaches highlights the importance of selecting the right approach for accurate, corrected measurement. The internal regression correction approach is appropriate for epidemiological studies because it takes into account inflammation severity. However, other models should be explored that account for inflammation and also provide better adjusted ferritin concentrations.
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Anemia Ferropriva , Anemia Ferropriva/diagnóstico , Anemia Ferropriva/epidemiologia , Biomarcadores , Pré-Escolar , Cuba/epidemiologia , Humanos , Lactente , Inflamação/epidemiologia , Ferro , Estado Nutricional , Orosomucoide/análise , PrevalênciaRESUMO
Alzheimer disease (AD) is the primary form of dementia that occurs spontaneously in older adults. Interestingly, the epigenetic profile of the cells forming the central nervous system changes during aging and may contribute to the progression of some neurodegenerative diseases such as AD. In this review, we present general insights into relevant epigenetic mechanisms and their relationship with aging and AD. The data suggest that some epigenetic changes during aging could be utilized as biomarkers and target molecules for the prevention and control of AD.
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Cigarette smoking has been associated with an increase in oxidative stress (OS) and is considered a predisposing factor to chronic noncommunicable diseases, whilst dietary antioxidants has been proposed as an alternative to cope with this oxidative stress. In this study, 20 smokers and 20 non-smokers were studied with the aim of determining their antioxidant status, as well as the ability of an infusion of 23 medicinal plants, to counteract the damage caused by OS. The plasma, red blood cells (RBCs) and polymorphonuclear cells (PBMCs) of both groups were incubated or not with the horchata infusion extract and then the OS markers, genotoxicity, nanostructure of RBCs membrane and genes related to oxidative responses and cellular functionality were evaluated. Up to 33 different compounds, mainly quercetin glycosides, were identified in the extract. A significant deterioration in the antioxidant status in smokers compared to non-smokers was found. The horchata infusion extract improved the nanostructure of RBCs and DNA damage, as well as the activity of the endogenous antioxidant enzymes and markers of oxidative damage to lipid, and proteins in plasma, RBCs and PBMCs in both groups, whilst no significant changes were found in the expression of different genes related to OS response.
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Bebidas , Fumar Cigarros/efeitos adversos , Estresse Oxidativo/efeitos dos fármacos , Plantas Medicinais , Adulto , Biomarcadores/sangue , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Fumantes , Adulto JovemRESUMO
AIM: VITAL, a phase II single-arm study, aimed to evaluate efficacy and safety of panitumumab addition to 5-fluorouracil (5-FU), mitomycin-C (MMC) and radiotherapy (RT) in patients with localized squamous cell carcinoma of the anal canal (SCCAC). METHODS: Adult, treatment-naïve SCCAC patients (Stage T2-T4, any N, M0) and ECOG-PS ≤2, received panitumumab (6 mg/kg, day 1 and Q2W; 8 weeks), 5-FU (1000 mg/m2 /d, days 1-4 and 29-32), MMC (10 mg/m2 , days 1 and 29) and RT 45 Gy (1.8 Gy/fraction) to the primary tumor and mesorectal, iliac and inguinal lymph nodes, plus 10-15 Gy boost dose to the primary tumor and affected lymph nodes. The primary objective was disease free survival rate (DFS) at 3-years (expected 3-year DFS rate: 73.7 ± 12%). RESULTS: Fifty-eight patients (31 women; median age: 59 years; ECOG-PS 0-1:98%; TNM II [29%] (T2 or T3/N0/M0)/IIIA (T1-T3/N1/M0 or T4/N0/M0) [21%]/IIIB (T4/N1/M0 or any T/N2 or N3/M0) [47%]/nonevaluable [4%]) were included. The median follow-up was 45 months. The 3-year DFS rate was 61.1% (95% CI: 47.1, 72.4). The 3-year overall survival rate was 78.4% (95% CI: 65.1, 87.1). Eighteen patients (31.0%) required a colostomy within 2 years posttreatment. Grade 3-4 toxicities were experienced by 53 (91%) patients. Most common grade 3-4 treatment-related events were radiation skin injury (40%) and neutropenia (24%). No toxic deaths occurred. Improved efficacy in colostomy-free survival and complete response rate was observed in human papilloma virus positive patients. CONCLUSIONS: Panitumumab addition to MMC-5FU regimen in SCCAC patients increases toxicity and does not improve patients' outcomes. RT plus MMC-5FU remains the standard of care for localized SCCAC patients.
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Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Neoplasias do Ânus/terapia , Quimiorradioterapia Adjuvante/efeitos adversos , Terapia Neoadjuvante/efeitos adversos , Neutropenia/epidemiologia , Radiodermite/epidemiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Neoplasias do Ânus/mortalidade , Quimiorradioterapia Adjuvante/métodos , Intervalo Livre de Doença , Feminino , Fluoruracila/administração & dosagem , Fluoruracila/efeitos adversos , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Mitomicina/administração & dosagem , Mitomicina/efeitos adversos , Terapia Neoadjuvante/métodos , Neutropenia/diagnóstico , Neutropenia/etiologia , Panitumumabe/administração & dosagem , Panitumumabe/efeitos adversos , Protectomia , Radiodermite/diagnóstico , Radiodermite/etiologia , Índice de Gravidade de Doença , Taxa de SobrevidaRESUMO
Trichomonas vaginalis is the etiological agent of human trichomoniasis. Metronidazole has high treatment success rate among trichomoniasis patients. However, metronidazole-resistant T. vaginalis has been reported, contributing in an increasing number of refractory cases. The mechanism of metronidazole resistance in this parasite is still unclear. In the vaginal environment, where the microaerophilic conditions prevail but the iron concentration is constantly fluctuating, the metronidazole resistance profile of T. vaginalis could be altered. In this study, we developed metronidazole-resistant strains of T. vaginalis and evaluate if iron availability is important to the action of the drug. The modulation of iron levels and iron chelation affected the actions of metronidazole both in susceptible and resistant strains. Interestingly, the early resistant strains exhibited minor iron content. The results of transcription analysis in the early resistant strains showed dysregulation in the expression of genes that codified proteins involved in iron transporter, iron-sulfur cluster assemblage, and oxidative stress response, which could not be observed in the late resistant and susceptible strains. Our results indicate that iron content plays an important role in the metronidazole action in T. vaginalis and likely to be related to iron-sulfur proteins involved in metronidazole activation and oxidative stress via Fenton reaction.
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Antiprotozoários/farmacologia , Resistência a Medicamentos/fisiologia , Ferro/metabolismo , Metronidazol/farmacologia , Trichomonas vaginalis/efeitos dos fármacos , Feminino , Humanos , Vaginite por Trichomonas , Trichomonas vaginalis/fisiologiaRESUMO
In our country, it is estimated that about 40% of the female population are physically and/or mentally abused by their partners, and between 21% and 60% of those women develop post-traumatic stress disorder. In this population, the knowledge about coping strategies is limited. OBJECTIVES: Identify whether signifcant differences exist as regards coping behaviors used in victims of gender violence between those women who develop post-traumatic stress disorder and those who do not. METHODS: The study involved 30 women who received an informed consent, a sociodemographic questionnaire, the questionnaire of the coping responses inventory CRI-A and the SCID-I. RESULTS: There were statistically signifcant differences regarding the use of logical analysis, seeking guidance and support, cognitive avoidance, and acceptance-resignation, used more by the post-traumatic stress disorder group; and minor use of positive reappraisal, problem solving and emotional discharge. DISCUSSION: Coping strategies of positive revaluation, problem solving and emotional discharge have been related to the decrease in the severity of depressive and anxious symptomatology. On the other hand, the usage of Cognitive Avoidance has been related to the increase in anxious and depressive symptomatology.
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Adaptação Psicológica , Transtornos de Estresse Pós-Traumáticos , Violência , Ansiedade , Feminino , Humanos , Inquéritos e Questionários , Violência/psicologiaRESUMO
BACKGROUND: Psychological and educational researchers are experiencing many practical difficulties in following the guidelines of the American Psychological Association (APA) for their statistical analyses: one such difficulty is the reporting of an effect-size measure along with each test of statistical significance (APA, 2010). The problem is exacerbated when researchers focus on contrast analysis instead of omnibus tests and when the Type-I error rate per comparison has to be adjusted. METHOD: Several reasons for this problem are discussed, with emphasis on the facts that researchers may be presented with too many optional effect-size measures with varying degrees of adequacy in several designs, and common statistical packages fail to provide appropriate effect-size measures for contrast analysis. RESULTS: This study proposes specific procedures (also implemented in spreadsheets) to compute generalized eta squared for various kinds of hypotheses, either general or specific, for one-factor and factorial between-group designs, and with manipulated and/or measured factors. CONCLUSIONS: Finally, conclusions are drawn concerning the need to take into account the kind of design and the kind of hypothesis in order to calculate comparable effect-size indexes across different types of studies and to prevent an overestimation of effect size.
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Interpretação Estatística de Dados , Projetos de Pesquisa/estatística & dados numéricos , Guias como Assunto , Psicologia/estatística & dados numéricos , Tamanho da AmostraRESUMO
Arginine kinase (AK) (ATP: L-arginine phosphotransferase, E.C. 2.7.3.3) catalyzes the reversible transfer of ATP γ-phosphate group to L-arginine to synthetize phospho-arginine as a high-energy storage. Previous studies suggest additional roles for AK in cellular processes. Since AK is found only in invertebrates and it is homologous to creatine kinase from vertebrates, the objective of this work was to demonstrate nucleoside diphosphate kinase-like activity for shrimp AK. For this, AK from marine shrimp Litopenaeus vannamei (LvAK) was purified and its activity was assayed for phosphorylation of TDP using ATP as phosphate donor. Moreover, by using high-pressure liquid chromatography (HPLC) the phosphate transfer reaction was followed. Also, LvAK tryptophan fluorescence emission changes were detected by dTDP titration, suggesting that the hydrophobic environment of Trp 221, which is located in the top of the active site, is perturbed upon dTDP binding. The kinetic constants for both substrates Arg and dTDP were calculated by isothermal titration calorimetry (ITC). Besides, docking calculations suggested that dTDP could bind LvAK in the same cavity where ATP bind, and LvAK basic residues (Arg124, 126 and 309) stabilize the dTDP phosphate groups and the pyrimidine base interact with His284 and Ser122. These results suggest that LvAK bind and phosphorylate dTDP being ATP the phosphate donor, thus describing a novel alternate nucleoside diphosphate kinase-like activity for this enzyme.
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Arginina Quinase/metabolismo , Núcleosídeo-Difosfato Quinase/metabolismo , Penaeidae/enzimologia , Nucleotídeos de Timina/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Simulação de Acoplamento Molecular , FosforilaçãoRESUMO
Conformational and thermal-rheological properties of acidic (APC) and neutral (NPC) protein concentrates were evaluated and compared to those of squid (Dosidicus gigas) muscle proteins (SM). Surface hydrophobicity, sulfhydryl status, secondary structure profile, differential scanning calorimetry and oscillatory dynamic rheology were used to evaluate the effect of treatments on protein properties. Acidic condition during the washing process (APC) promoted structural and conformational changes in the protein present in the concentrate produced. These changes were enhanced during the heat setting of the corresponding sol. Results demonstrate that washing squid muscle under the proposed acidic conditions is a feasible technological alternative for squid-based surimi production improving its yield and gel-forming ability.
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Decapodiformes/química , Manipulação de Alimentos/métodos , Proteínas Musculares/química , Alimentos Marinhos/análise , Animais , Varredura Diferencial de Calorimetria , Concentração de Íons de Hidrogênio , Músculos/química , ReologiaRESUMO
The tons of organic waste that are annually generated by agro-industry, can be used as raw material for methane production. For this reason, it is important to previously perform biodegradability tests to organic wastes for their full scale methanization. This paper addresses biodegradability, methane production and the behavior of populations of eubacteria and archaeabacteria during anaerobic digestion of banana, mango and papaya agroindustrial wastes. Mango and banana wastes had higher organic matter content than papaya in terms of their volatile solids and total solid rate (94 and 75% respectively). After 63 days of treatment, the highest methane production was observed in banana waste anaerobic digestion: 63.89ml CH4/per gram of chemical oxygen demand of the waste. In the PCR-DGGE molecular analysis, different genomic footprints with oligonucleotides for eubacteria and archeobacteria were found. Biochemical methane potential results proved that banana wastes have the best potential to be used as raw material for methane production. The result of a PCR- DGGE analysis using specific oligonucleotides enabled to identify the behavior of populations of eubacteria and archaeabacteria present during the anaerobic digestion of agroindustrial wastes throughout the process.
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Agricultura , Anaerobiose , Archaea/isolamento & purificação , Bactérias/isolamento & purificação , Biocombustíveis , Reatores Biológicos , Resíduos Industriais , Metano/biossíntese , Resíduos Sólidos , Archaea/metabolismo , Bactérias/metabolismo , Biodegradação Ambiental , Carica , DNA Bacteriano/análise , Mangifera , Metano/isolamento & purificação , Plantago , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Eliminação de Resíduos , RibotipagemRESUMO
Las toneladas de residuos orgánicos que se generan anualmente en la agroindustria pueden aprovecharse como materia prima para la producción de metano. Para que los residuos orgánicos se puedan convertir a metano a gran escala, es importante que previamente se realicen sobre ellos pruebas de biodegradabilidad; un parámetro importante que conviene establecer es su potencial bioquímico de metano. En el presente trabajo se estudió la biodegradabilidad, la producción de metano y el comportamiento de poblaciones de eubacterias y arqueobacterias durante la digestión anaerobia de residuos de plátano, mango y papaya provenientes de la agroindustria, adicionando un inóculo microbiano. Los residuos de mango y plátano tenían mayor contenido de materia orgánica (94 y 75 %, respectivamente) que el residuo de papaya con base en su relación sólidos volátiles/sólidos totales. Después de 63 días de tratamiento, la mayor producción de metano se observó en la digestión anaerobia del residuo de plátano: 63,89 ml de metano por g de demanda química de oxígeno del residuo. Los resultados del potencial bioquímico de metano demostraron que el residuo de plátano tiene el mejor potencial para ser usado como materia prima en la producción de metano. A través de un análisis por PCR-DGGE con oligonucleótidos específicos se logró evaluar el tamaño y la composición de las poblaciones de eubacterias y arqueobacterias presentes en la digestión anaerobia de residuos agroindustriales a lo largo del proceso.
The tons of organic waste that are annually generated by agro-industry, can be used as raw material for methane production. For this reason, it is important to previously perform biodegradability tests to organic wastes for their full scale methanization. This paper addresses biodegradability, methane production and the behavior of populations of eubacteria and archaeabacteria during anaerobic digestion of banana, mango and papaya agroindustrial wastes. Mango and banana wastes had higher organic matter content than papaya in terms of their volatile solids and total solid rate (94 and 75 % respectively). After 63 days of treatment, the highest methane production was observed in banana waste anaerobic digestion: 63.89 ml CH4/per gram of chemical oxygen demand of the waste. In the PCR-DGGE molecular analysis, different genomic footprints with oligonucleotides for eubacteria and archeobacteria were found. Biochemical methane potential results proved that banana wastes have the best potential to be used as raw material for methane production. The result of a PCR- DGGE analysis using specific oligonucleotides enabled to identify the behavior of populations of eubacteria and archaeabacteria present during the anaerobic digestion of agroindustrial wastes throughout the process.
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Digestão Anaeróbia/métodos , Gerenciamento de Resíduos/métodos , Metano/biossíntese , Biodegradação Ambiental , Uso de Resíduos Sólidos , Agroindústria/prevenção & controle , Reciclagem/métodos , Análise da Demanda Biológica de Oxigênio/métodos , Resíduos de AlimentosRESUMO
Nucleotide phosphorylation is a key step in DNA replication and viral infections, since suitable levels of nucleotide triphosphates pool are required for this process. Deoxythymidine monophosphate (dTMP) is produced either by de novo or salvage pathways, which is further phosphorylated to deoxythymidine triphosphate (dTTP). Thymidyne monophosphate kinase (TMK) is the enzyme in the junction of both pathways, which phosphorylates dTMP to yield deoxythymidine diphosphate (dTDP) using adenosine triphosphate (ATP) as a phosphate donor. White spot syndrome virus (WSSV) genome contains an open reading frame (ORF454) that encodes a thymidine kinase and TMK domains in a single polypeptide. We overexpressed the TMK ORF454 domain (TMKwssv) and its specific activity was measured with dTMP and dTDP as phosphate acceptors. We found that TMKwssv can phosphorylate dTMP to yield dTDP and also is able to use dTDP as a substrate to produce dTTP. Kinetic parameters K M and k cat were calculated for dTMP (110 µM, 3.6 s(-1)), dTDP (251 µM, 0.9 s(-1)) and ATP (92 µM, 3.2 s(-1)) substrates, and TMKwssv showed a sequential ordered bi-bi reaction mechanism. The binding constants K d for dTMP (1.9 µM) and dTDP (10 µM) to TMKwssv were determined by Isothermal Titration Calorimetry. The affinity of the nucleotidic analog stavudine monophosphate was in the same order of magnitude (K d 3.6 µM) to the canonical substrate dTMP. These results suggest that nucleotide analogues such as stavudine could be a suitable antiviral strategy for the WSSV-associated disease.
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Núcleosídeo-Fosfato Quinase/química , Fases de Leitura Aberta , Proteínas Virais/química , Vírus da Síndrome da Mancha Branca 1/enzimologia , Núcleosídeo-Fosfato Quinase/antagonistas & inibidores , Núcleosídeo-Fosfato Quinase/genética , Estrutura Terciária de Proteína , Especificidade por Substrato/fisiologia , Proteínas Virais/antagonistas & inibidores , Proteínas Virais/genética , Vírus da Síndrome da Mancha Branca 1/genéticaRESUMO
Overweight and obesity are associated with systemic inflammation and oxidative stress which, in turn, enhance the development of cardiometabolic disruptions. Lifestyle changes and pharmacologic approaches show moderately effective results regarding overall health improvements. Evidence suggests that cacao flavonoids are associated with a reduced cardiometabolic risk, due to the modulation of molecular pathways subjacent to glucose and lipids metabolism. The aim of this study was to assess the effects of cacao flavonoids supplementation on anthropometric and cardiometabolic risk factors in overweight subjects. A double-blind, placebo-controlled, pilot clinical trial was conducted in overweight subjects with borderline criteria of metabolic syndrome. Participants were randomly assigned to either, supplement of cacao flavonoids (80 mg) or placebo, daily, for 4 weeks. Cardiometabolic variables were blood pressure, glycemia and lipid profile. Serum markers of oxidative damage (free protein carbonyls and malondialdehyde) were also analyzed. Anthropometric measurements included body weight, body mass index, waist circumference, and fat and fat-free mass. We found significant reductions in body weight (p = 0.04), waist circumference (p = 0.03), triacylglycerols (p < 0.01), TG/HDL ratio (p = 0.01), MDA (p = 0.02) and protein carbonyls (p = 0.01) in the flavonoid-supplemented group. Results from this study show that cacao flavonoids can effectively modulate anthropometric and cardiometabolic risk factors.
El sobrepeso y la obesidad están asociados con la inflamación sistémica y el estrés oxidativo, que, a su vez, incrementan el desarrollo de trastornos cardiometabólicos. Cambios en el estilo de vida y tratamientos farmacológicos muestran resultados moderadamente eficaces en relación con la mejora general de la salud. La evidencia sugiere que los flavonoides del cacao se asocian con un riesgo cardiometabólico reducido, debido a la modulación de las vías moleculares subyacentes al metabolismo de la glucosa y de los lípidos. El objetivo de este estudio fue evaluar los efectos de la suplementación de flavonoides del cacao sobre factores de riesgo cardiometabólico y antropométrico en sujetos con sobrepeso. Se llevó a cabo un ensayo clínico piloto, doble ciego y controlado con placebo en sujetos con sobrepeso y criterios limítrofes de síndrome metabólico. Los participantes fueron asignados al azar a cuatro semanas de tratamiento con suplemento oral de flavonoides de cacao (80 mg) diario o placebo. Las variables cardiometabólicas analizadas fueron presión arterial sistémica, glicemia y perfil lipídico. También se analizaron los marcadores séricos de estrés oxidativo (carbonilos proteicos libres y malondialdehído). Las medidas antropométricas incluyeron el peso corporal, índice de masa corporal, circunferencia de la cintura, masa grasa y masa libre de grasa. Se encontró una reducción significativa en el peso corporal (p = 0.04), circunferencia de la cintura (p = 0.03), triglicéridos (p < 0.01), la relación TG/HDL (p = 0.01), MDA (p = 0.02) y carbonilos (p = 0.01) en el grupo con suplemento de flavonoides. Los resultados de este estudio muestran que los flavonoides del cacao pueden modular efectivamente factores de riesgo cardiometabólico y antropométricos.
RESUMO
BACKGROUND: Coronary flow velocity reserve (CFVR) increases the diagnostic sensitivity of stress echocardiography. The aim of this study was to evaluate the prognostic value of CFVR in patients without new wall motion abnormalities during pharmacologic stress echocardiography. METHODS: The outcomes of 651 patients with normal wall motion response during stress echocardiography with dobutamine up to 50 µg/kg/min (n = 351) or dipyridamole up to 0.84 mg/kg over 4 min (n = 300) were evaluated. CFVR was calculated simultaneously in the distal territory of the left anterior descending coronary artery. CFVR ≥ 2 was defined as normal. Major events considered during follow-up were cardiovascular death, myocardial infarction, and late myocardial revascularization. RESULTS: Normal CFVR was recorded in 523 patients and reduced CFVR in 128. During a mean follow-up period of 34.6 ± 18 months, 48 major events occurred, in 25 patients (4.8%) with normal and 23 patients (18%) with reduced CFVR; event-free survival was significantly different between patients with normal versus abnormal CFVR (P < .0001). Diabetes increased risk only in patients with abnormal CFVR (P = .05). In the multivariate analysis, CFVR and history of smoking were the only independent predictors of combined morbidity and mortality. Abnormal CFVR was associated with a higher event rate, independently of the pharmacologic stress technique used. The event hazard ratio was inversely proportional to CFVR. CONCLUSIONS: CFVR was an independent predictor of mortality after pharmacologic stress echocardiography with normal wall motion, and the degree of decrease was associated with increased risk. Diabetes worsened prognosis only with abnormal CFVR.
Assuntos
Estenose Coronária/mortalidade , Estenose Coronária/fisiopatologia , Complicações do Diabetes/mortalidade , Complicações do Diabetes/fisiopatologia , Dobutamina , Ecocardiografia/estatística & dados numéricos , Reserva Fracionada de Fluxo Miocárdico , Idoso , Argentina/epidemiologia , Comorbidade , Estenose Coronária/diagnóstico por imagem , Complicações do Diabetes/diagnóstico por imagem , Teste de Esforço/métodos , Feminino , Humanos , Masculino , Contração Miocárdica/efeitos dos fármacos , Prevalência , Prognóstico , Reprodutibilidade dos Testes , Fatores de Risco , Sensibilidade e Especificidade , Taxa de Sobrevida , VasodilatadoresRESUMO
Several proteases from plant sources have been proposed as milk coagulants, however, limited research has been done on their milk-clotting properties. The effect of temperature on the milk-clotting activity of kiwi fruit, melon and ginger extracts was evaluated, as well as the effects of the different extracts on curd properties. Melon extracts showed high milk-clotting activity over a broad temperature range (45-75 °C) while kiwi fruit and ginger extracts showed high activity over a narrower temperature range, with a maximum at 40 and 63 °C, respectively. Curds produced using kiwi extracts had textural properties comparable with those obtained using commercial rennet, while melon extracts produced a fragile gel and low curd yield. The milk-clotting behavior of the three plant extracts was related to the protease specificity present in these extracts. The kiwi proteases displayed chymosin-like properties and thus hold the best potential for use as a milk coagulant in cheese production.
Assuntos
Actinidia/química , Queijo/análise , Cucurbitaceae/química , Aditivos Alimentares/química , Leite/química , Extratos Vegetais/química , Zingiber officinale/química , Animais , Bovinos , Concentração de Íons de Hidrogênio , TemperaturaRESUMO
Jumbo squid (Dosidicus gigas) mantle muscle was cooked simulating industrial procedures (95 degrees C x 25 min, 1.2:5 muscle:water ratio). The effluent produced was analyzed for chemical and biochemical oxygen demands (COD and BOD(5), respectively), proximate analysis, flavor-related compounds (free amino acids, nucleotides and carbohydrates) and SDS-PAGE. The COD and BOD(5) exhibited variation among samplings (N=3) (27.4-118.5 g O(2)/L for COD and 11.3-26.7 g O(2)/L for BOD(5)). The effluent consisted of 1% total solids, 75% of which represented crude protein. Sixty percent of the total free amino acid content, which imparts flavor in squid species, corresponded to glutamic acid, serine, glycine, arginine, alanine, leucine and lysine. The nucleotide concentration followed this order, Hx>>ADP>AMP>ATP>IMP>HxR. The variation observed in the present work was probably due to physiological maturity differences among the squid specimens (i.e., juvenile versus mature). Solids present in squid cooking effluent could be recovered and potentially used as flavor ingredients in squid-analog production by the food industry.