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BACKGROUND: The ubiquitin-proteasome system (UPS) and autophagy are 2 major protein degradation pathways in eukaryotic cells. We previously identified a switch from UPS to autophagy with changes in BAG3 (B-cell lymphoma 2-associated-athanogene 3) expression after cerebral ischemia in mice. BAG3 is an antiapoptotic-cochaperone that is directly involved in cellular protein quality control as a mediator for selective macroautophagy. Here, we aimed to investigate the role of BAG3 in ischemic stroke. METHODS: Middle cerebral artery occlusion/reperfusion (MCAO/R) and oxygen-glucose deprivation/reoxygenation were used to mimic cerebral ischemia in vivo and in vitro. The UPS inhibitor MG132 and autophagy inhibitor 3-MA (3-methyladenine) were administered to mice to identify how BAG3 was involved after MCAO/R. Adeno-associated virus and lentiviral vector were used to regulate BAG3 expression in vivo and in vitro, respectively. Behavioral tests, 2,3,5-triphenyltetrazolium chloride staining, and Hematoxylin & Eosin staining were performed to evaluate cerebral injury following MCAO/R, and a Cell Counting kit-8 assay was conducted to assess oxygen-glucose deprivation/reoxygenation-induced injury in cells. Brain tissues and cell lysates were collected and analyzed for UPS activation, autophagy, and apoptosis. RESULTS: The UPS inhibitor alleviated MCAO injury in mice and increased autophagy and BAG3 expression, whereas the autophagy inhibitor exacerbated MCAO/R-induced injury. In addition, BAG3 overexpression significantly improved neurological outcomes, reduced infarct volume in vivo, and enhanced cell survival by activating autophagy and suppressing apoptosis in vitro. CONCLUSIONS: Our findings indicate that BAG3 overexpression activates autophagy and inhibits apoptosis to prevent cerebral ischemia/reperfusion and hypoxia/reoxygenation injury, suggesting a potential therapeutic benefit of BAG3 expression in cerebral ischemia.
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Isquemia Encefálica , Accidente Cerebrovascular Isquémico , Daño por Reperfusión , Animales , Ratones , Apoptosis , Autofagia , Isquemia Encefálica/metabolismo , Glucosa , Infarto de la Arteria Cerebral Media , Oxígeno , Daño por Reperfusión/metabolismoRESUMEN
OBJECTIVE: Exaggerated exercise blood pressure (BP) is associated with cardiovascular risk factors in adolescence. Cardiorespiratory fitness and adiposity (fatness) are independent contributors to cardiovascular risk, but their interrelated associations with exercise BP are unknown. This study aimed to determine the relationships between fitness, fatness, and the acute BP response to exercise in a large birth cohort of adolescents. METHODS: 2292 adolescents from the Avon Longitudinal Study of Parents and Children (aged 17.8 ± 0.4 years, 38.5% male) completed a sub-maximal exercise step test that allowed fitness (VO2 max ) to be determined from workload and heart rate using a validated equation. Exercise BP was measured immediately on test cessation and fatness calculated as the ratio of total fat mass to total body mass measured by DXA. RESULTS: Post-exercise systolic BP decreased stepwise with tertile of fitness (146 (18); 142 (17); 141 (16) mmHg) but increased with tertile of fatness (138 (15); 142 (16); 149 (18) mmHg). In separate models, fitness and fatness were associated with post-exercise systolic BP adjusted for sex, age, height, smoking, and socioeconomic status (standardized ß: -1.80, 95%CI: -2.64, -0.95 mmHg/SD and 4.31, 95%CI: 3.49, 5.13 mmHg/SD). However, when fitness and fatness were included in the same model, only fatness remained associated with exercise BP (4.65, 95%CI: 3.69, 5.61 mmHg/SD). CONCLUSION: Both fitness and fatness are associated with the acute BP response to exercise in adolescence. The fitness-exercise BP association was not independent of fatness, implying the cardiovascular protective effects of cardiorespiratory fitness may only be realized with more favorable body composition.
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Adiposidad/fisiología , Presión Sanguínea/fisiología , Capacidad Cardiovascular/fisiología , Ejercicio Físico/fisiología , Adolescente , Estudios Transversales , Prueba de Esfuerzo , Femenino , Humanos , Estudios Longitudinales , MasculinoRESUMEN
PURPOSE: Exaggerated exercise blood pressure (BP) is associated with altered cardiac structure and increased cardiovascular risk. Fitness modifies these associations, but the effect in healthy adolescents is unknown. We performed an observational study to determine the influence of fitness on post-exercise BP, and on its relationship with cardiac structure in adolescents. METHODS: 4835 adolescents from the Avon Longitudinal Study of Parents and Children, (15.4 (0.3) years, 49% male) completed a submaximal cycle test. Fitness was estimated as physical work capacity 170 adjusted for lean body mass and post-exercise BP measured immediately posttest. Cardiovascular structure and function, including left ventricular (LV) mass (n = 1589), left atrium (LA) size (n = 1466), cardiac output (CO, n = 1610), and total peripheral resistance (TPR, n = 1610) were measured at rest by echocardiography 2.4 (0.4) years later. RESULTS: Post-exercise systolic BP increased stepwise by fitness tertile (131.2 mm Hg [130.4, 132.1]; 137.3 mm Hg [136.5, 138.0]; 142.3 mm Hg [141.5, 143.1]). Each 5 mm Hg of post-exercise systolic BP was associated with 2.46 g [1.91, 3.01] greater LV mass, 0.02 cm [0.02, 0.03] greater LA size, and 0.25 g/m2.7 [0.14, 0.36] greater LV mass index. Adjustment for fitness abolished associations (0.29 g [-0.16, 0.74]; 0.01 cm [-0.001, 0.014] and 0.08 g/m2.7 [-0.001, 0.002]). Similar associations between post-exercise systolic BP and each outcome were found between the lowest and highest fitness thirds. CO increased with fitness third (difference 0.06 L/min [-0.05, 0.17]; 0.23 L/min [0.12, 0.34]) while TPR decreased (difference -0.13 mm Hg·min/L [-0.84,0.59]; -1.08 mm Hg·min/L [-0.1.80, 0.35]). CONCLUSIONS: Post-exercise systolic BP increased with fitness, which modified its association with cardiac structure. Higher CO, but lower TPR suggests a physiologically adapted cardiovascular system with greater fitness, highlighting the importance of fitness in adolescence.
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Presión Sanguínea , Ejercicio Físico , Corazón/fisiología , Aptitud Física , Adolescente , Ecocardiografía , Prueba de Esfuerzo , Femenino , Voluntarios Sanos , Corazón/diagnóstico por imagen , Humanos , Estudios Longitudinales , MasculinoRESUMEN
PURPOSE: Masked hypertension is associated with increased cardiovascular risk but is undetectable by clinic blood pressure (BP). Elevated systolic BP responses to submaximal exercise reveal the presence of masked hypertension in adults, but it is unknown whether this is the case during adolescence. We aimed to determine if exercise BP was raised in adolescents with masked hypertension, and its association with cardiovascular risk markers. METHODS: A total of 657 adolescents (aged 17.7 ± 0.3 years; 41.9% male) from the Avon longitudinal study of parents and children (ALSPAC) completed a step-exercise test with pre-, post-, and recovery-exercise BP, clinic BP and 24-hour ambulatory BP. Masked hypertension was defined as clinic BP <140/90 mm Hg and 24-hour ambulatory BP ≥130/80 mm Hg. Assessment of left-ventricular (LV) mass index and carotid-femoral pulse wave velocity (aortic PWV) was also undertaken. Thresholds of clinic, pre-, post-, and recovery-exercise systolic BP were explored from ROC analysis to identify masked hypertension. RESULTS: Fifty participants (7.8%) were classified with masked hypertension. Clinic, pre-, post-, and recovery-exercise systolic BP were associated with masked hypertension (AUC ≥ 0.69 for all, respectively), with the clinic systolic BP threshold of 115 mm Hg having high sensitivity and specificity and exercise BP thresholds of 126, 150, and 130 mm Hg, respectively, having high specificity and negative predictive value (individually or when combined) for ruling out the presence of masked hypertension. Additionally, this exercise systolic BP above the thresholds was associated with greater left-ventricular mass index and aortic PWV. CONCLUSIONS: Submaximal exercise systolic BP is associated with masked hypertension and adverse cardiovascular structure in adolescents. Exercise BP may be useful in addition to clinic BP for screening of high BP and cardiovascular risk in adolescents.
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Presión Sanguínea , Ejercicio Físico , Hipertensión Enmascarada/diagnóstico , Adolescente , Monitoreo Ambulatorio de la Presión Arterial , Prueba de Esfuerzo , Femenino , Humanos , Estudios Longitudinales , Masculino , Análisis de la Onda del Pulso , Sensibilidad y Especificidad , SístoleRESUMEN
Cardiovascular disease remains the leading cause of mortality on a global scale. Individuals who possess risk factors for cardiovascular disease, such as high blood pressure (BP) and obesity, face an elevated risk of experiencing organ-specific pathophysiological changes. This damage includes pathophysiological changes in the heart and peripheral vascular systems, such as ventricular hypertrophy, arterial stiffening, and vascular narrowing and stenosis. Consequently, these damages are associated with an increased risk of developing severe cardiovascular outcomes including stroke, myocardial infarction, heart failure, and coronary heart disease. Among all the risk factors associated with cardiovascular disease, high blood pressure emerges as the most prominent. However, conventional resting BP measurement methods such as auscultatory or oscillometric methods may fail to identify many individuals with asymptomatic high BP. Recently, exercise BP has emerged as a valuable diagnostic tool for identifying real (high) blood pressure levels and assessing underlying cardiovascular risk, in addition to resting BP measurements in adults. Furthermore, numerous established factors, such as low cardiorespiratory fitness and high body fatness, have been confirmed to contribute to exercise BP and the associated cardiovascular risk. Modifying these factors may help reduce high exercise BP and, consequently, alleviate the burden of cardiovascular disease. A significant body of evidence has demonstrated cardiovascular disease in later life have their origins in early life. Children and adolescents with these cardiovascular risk factors also possess a greater propensity to develop cardiovascular diseases later in life. Nevertheless, the majority of previous studies on the clinical utility of exercise BP have been conducted in middle-to-older aged populations, often with pre-existing clinical conditions. Therefore, there is a need to investigate further of the factors influencing exercise BP in adolescence and its association with cardiovascular risk in early life. Our previously published work showed that exercise BP is a potential useful method to detect adolescents with increased cardiovascular risk. Children and adolescents with cardiovascular risk factors are more likely to develop cardiovascular diseases later in life. However, previous studies on the clinical utility of exercise BP have largely focused on middle-to-older aged populations with pre-existing clinical conditions. Therefore, there is a need to investigate further the factors influencing exercise BP in adolescence and its association with future cardiovascular risk. Our previous studies, which focused on exercise BP measured at submaximal intensity, have shown that exercise BP is a potentially useful method for identifying adolescents at increased cardiovascular risk. Our previous findings suggest that improving cardio-respiratory fitness and reducing body fatness may help to reduce the risk of developing cardiovascular disease and improve overall cardiovascular health. These findings have important implications for the development of effective prevention and early detection strategies, which can contribute to improved public health outcomes.
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Presión Sanguínea , Capacidad Cardiovascular , Enfermedades Cardiovasculares , Ejercicio Físico , Humanos , Niño , Adolescente , Capacidad Cardiovascular/fisiología , Presión Sanguínea/fisiología , Factores de Riesgo , Masculino , Factores de Riesgo de Enfermedad Cardiaca , FemeninoRESUMEN
As the resident immune cells in the central nervous system, microglia exhibit a 'sensitized' or 'primed' phenotype with dystrophic morphology and dysregulated functions in aged brains. Although studies have demonstrated the inflammatory profile of aged microglia in several neurological diseases, this issue is largely uncertain in stroke. Consequently, this study investigated the effects of primed and repopulated microglia on post-ischemic brain injury in aged mice. We replaced primed microglia with newly repopulated microglia through pharmacological administration and withdrawal of the colony-stimulating factor 1 receptor (CSF1R) inhibitor, PLX3397. Further, we performed a series of behavioral tests and flow cytometry in mouse models of middle cerebral artery occlusion (MCAO) to study the effects of microglial replacement on ischemic injury in the aged brain. With depletion and subsequent repopulation of microglia in MCAO mice, microglial replacement in aged mice improved neurological function and decreased brain infarction. This protective effect was accompanied by the reduction of peripheral immune cells infiltrating into brains. We showed that the repopulated microglia expressed elevated neuroprotective factors (including Cluster of Differentiation 206, transforming growth factor-ß, and interleukin-10) and diminished expression of inflammatory markers (including Cluster of Differentiation 86, interleukin-6, and tumor necrosis factor α). Moreover, microglial replacement protected the blood-brain barrier and relieved neuronal death in aged mice subjected to 60 min of MCAO. These results imply that the replacement of microglia in the aged brain may alleviate brain damage and neuroinflammation, and therefore, ischemic brain damage. Thus, targeting microglia could be a promising therapeutic strategy for ischemic stroke.
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Envejecimiento , Infarto de la Arteria Cerebral Media , Ratones Endogámicos C57BL , Microglía , Enfermedades Neuroinflamatorias , Fármacos Neuroprotectores , Animales , Microglía/efectos de los fármacos , Microglía/inmunología , Masculino , Ratones , Infarto de la Arteria Cerebral Media/inmunología , Infarto de la Arteria Cerebral Media/patología , Infarto de la Arteria Cerebral Media/tratamiento farmacológico , Fármacos Neuroprotectores/uso terapéutico , Fármacos Neuroprotectores/farmacología , Enfermedades Neuroinflamatorias/tratamiento farmacológico , Enfermedades Neuroinflamatorias/inmunología , Enfermedades Neuroinflamatorias/patología , Aminopiridinas/farmacología , Aminopiridinas/uso terapéutico , Pirroles/farmacología , Pirroles/uso terapéutico , Isquemia Encefálica/tratamiento farmacológico , Encéfalo/patología , Encéfalo/efectos de los fármacos , Encéfalo/inmunología , Modelos Animales de Enfermedad , Receptores de Factor Estimulante de Colonias de Granulocitos y Macrófagos/antagonistas & inhibidores , Receptores de Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Barrera Hematoencefálica/efectos de los fármacosRESUMEN
How to avoid the microenvironment limitations in the therapeutic process of pressure ulcers is still challenging. The development of a functional gel can kill bacteria and scavenge reactive oxygen species (ROS), which is urgently required in the therapeutic process of pressure ulcers. Herein, an in situ sprayed gel is developed with silver nanoparticles (AgNPs) and polydopamine (PDA) NPs (APG) to obviate microenvironment restrictions in treating pressure ulcers. The gel is constructed by spraying sodium alginate solution and CaCl2 solution. AgNPs serve as an antibacterial agent in the formed gel, which can effectively cause bacterial inactivation and show more than 5 log (>99.999%) bacterial killing efficiency against methicillin-resistant S. aureus (MRSA), Staphylococcus aureus (S. aureus), and Escherichia coli (E. coli) in vitro. Meanwhile, PDA NPs serve as the antioxidative agent in the formed gel, which can facilitate the elimination of ROS to address the high ROS problem in wound microenvironment. Based on these features, it is demonstrated through cell and animal experiments that the AgNPs and PDA NPs incorporated gel can realize the effective treatment of MRSA-infected and hydrogen peroxide (H2 O2 )-sensitized pressure ulcers. It is believed that the designed system by a simple spray-coating approach can provide a new therapeutic strategy in biomedical areas.
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Nanopartículas del Metal , Staphylococcus aureus Resistente a Meticilina , Úlcera por Presión , Animales , Staphylococcus aureus , Úlcera por Presión/tratamiento farmacológico , Especies Reactivas de Oxígeno , Escherichia coli , Nanopartículas del Metal/uso terapéutico , Plata/farmacología , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Pruebas de Sensibilidad MicrobianaRESUMEN
With the popularity of computer-based testing (CBT), it is easier to collect item response times (RTs) in psychological and educational assessments. RTs can provide an important source of information for respondents and tests. To make full use of RTs, the researchers have invested substantial effort in developing statistical models of RTs. Most of the proposed models posit a unidimensional latent speed to account for RTs in tests. In psychological and educational tests, many tests are multidimensional, either deliberately or inadvertently. There may be general effects in between-item multidimensional tests. However, currently there exists no RT model that considers the general effects to analyze between-item multidimensional test RT data. Also, there is no joint hierarchical model that integrates RT and response accuracy (RA) for evaluating the general effects of between-item multidimensional tests. Therefore, a bi-factor joint hierarchical model using between-item multidimensional test is proposed in this study. The simulation indicated that the Hamiltonian Monte Carlo (HMC) algorithm works well in parameter recovery. Meanwhile, the information criteria showed that the bi-factor hierarchical model (BFHM) is the best fit model. This means that it is necessary to take into consideration the general effects (general latent trait) and the multidimensionality of the RT in between-item multidimensional tests.
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Background: The prospective association between plasma Se and stroke risk remains inconclusive. The relationship between Se and ischemic stroke among a low circulating Se status population deserves more attention, especially for Chinese people who were a high-risk group for Se deficiency. Objective: The relationship between plasma Se concentration and ischemic stroke risk in a large-scale Chinese community-based population and any potential effect modifiers were investigated. Methods: A nested, case-control study, using data from the "China H-type Hypertension Registry Study" were conducted. A total of 1,904 first ischemic stroke cases and 1,904 controls matched for age, sex, and village were included in this study. The association between plasma Se and first ischemic stroke was evaluated by conditional logistic regression analyses. Results: The median value of plasma Se was 65.8 µg/L among total participants. Overall, a significant inverse relationship between plasma Se and first ischemic stroke risk was found (per SD increment; adjusted OR: 0.87; 95% CI: 0.80 and 0.95). Accordingly, a significantly lower risk of first ischemic stroke was found in participants in quartile 3 (65.8-<77.8 µg/L) (adjusted OR: 0.78; 95% CI: 0.63 and 0.96) and quartile 4 (≥77.8 µg/L) (adjusted OR: 0.76; 95% CI: 0.59 and 0.96), compared with those in quartile 1 (<56.0 µg/L). Furthermore, a significantly lower ischemic stroke risk was found in those with lower low-density lipoprotein cholesterol (LDL-C) levels (<3.4 vs. ≥3.4 mmol/L; P for interaction = 0.015) or those with lower homocysteine levels (<12.1 (median) vs. ≥12.1 µmol/L; P for interaction = 0.027) at baseline. Conclusion: Plasma Se was significantly inversely associated with the risk of first ischemic stroke among a large-scale Chinese community-based population (most adults with hypertension and elevated total homocysteine), especially among those with lower LDL-C and lower homocysteine levels.
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Integrins αvß3 and αvß6 are highly expressed on tumor cells and/or by the tumor vasculature of many human cancers, and represent promising targets for anticancer therapy. Novel chemically programmed antibodies (cpAbs) targeting these integrins were prepared using the catalytic aldolase Antibody (Ab) programming strategy. The effects of the cpAbs on cellular functions related to tumor progression were examined in vitro using tumor cell lines and their cognate integrin ligands, fibronectin and osteopontin. The inhibitory functions of the conjugates and their specificity were examined based on interference with cell-cell and cell-ligand interactions related to tumor progression. Cell binding analyses of the anti-integrin cpAbs revealed high affinity for tumor cells that overexpressed αvß3 and αvß6 integrins, and weak interactions with αvß1 and αvß8 integrins, in vitro. Functional analyses demonstrated that the cpAbs strongly inhibited cell-cell interactions through osteopontin binding, and they had little or no immediate effects on cell viability and proliferation. On the basis of these characteristics, the cpAbs are likely to have a broad range of activities in vivo, as they can target and antagonize one or multiple αv integrins expressed on tumors and tumor vasculatures. Presumably, these conjugates may inhibit the establishment of metastastatic tumors in distant organs through interfering with cell adhesion more effectively than antibodies or compounds targeting one integrin only. These anti-integrin cpAbs may also provide useful reagents to study combined effect of multiple αv integrins on cellular functions in vitro, on pathologies, including tumor angiogenesis, fibrosis, and epithelial cancers, in vivo.
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Anticuerpos/uso terapéutico , Antígenos de Neoplasias/inmunología , Antineoplásicos , Integrina alfaV/inmunología , Neoplasias/inmunología , Anticuerpos/inmunología , Comunicación Celular , Línea Celular Tumoral , Fructosa-Bifosfato Aldolasa , Humanos , Inmunoconjugados/uso terapéutico , Integrina alfaVbeta3/inmunología , Integrinas/inmunología , Neoplasias/tratamiento farmacológicoRESUMEN
Chemical programming of nine murine antibodies with catalytic aldolase activity was examined using compounds, equipped with diketone or pro-vinyl ketone linkers that inhibit integrin adhesion receptor functions. The results showed that most Abs were programmed using the diketone compounds in a manner similar to previously reported catalytic antibody 38C2. On the other hand, only those antibodies, which catalyzed the retro aldol reaction of the pro-vinyl ketone linkers efficiently, were programmed. Conjugated to integrin targeting compounds, at least three new antibodies, including 84G3, 85H6, and 90G8, exhibited high specific binding to human tumor cells expressing integrin alpha(v)beta(3.).
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Anticuerpos Catalíticos/química , Fructosa-Bifosfato Aldolasa/química , Fragmentos Fab de Inmunoglobulinas/química , Anticuerpos Catalíticos/inmunología , Anticuerpos Catalíticos/metabolismo , Línea Celular Tumoral , Fructosa-Bifosfato Aldolasa/inmunología , Fructosa-Bifosfato Aldolasa/metabolismo , Humanos , Fragmentos Fab de Inmunoglobulinas/inmunología , Fragmentos Fab de Inmunoglobulinas/metabolismo , Integrina alfaVbeta3/metabolismo , Cetonas/químicaRESUMEN
The enhanced aerobic denitrification capability of the mixed strains YH01+YH02 in utilizing potassium nitrate, and the dynamic changes in the microbial community component during the period of operation, were evaluated. The microbial community in different stages of the SBR was analyzed by using high-throughput sequencing technology after inoculation with YH01+YH02. The results showed that the NO3--N, TN, and COD removal efficiencies increased by 12.1%, 9.2%, and 9.4%, respectively. The relative abundances of the microbes in the microbial community increased at the genus level, and the diversity in the microbial community decreased after enhancement. Principal component analysis and UPGMA analysis revealed that the period of SBR operation was roughly divined into four phases. The relative abundances of Delftia and Acidovorax increased during the period of operation, and YH01+YH02 exhibited excellent compatibility with the SBR ecosystem and played an important part in aerobic denitrification.
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Bacterias/clasificación , Reactores Biológicos/microbiología , Desnitrificación , Nitrógeno/aislamiento & purificación , Bacterias/metabolismoRESUMEN
To reveal the dynamic succession of microbial community structure along with time in bio-denitrification reactor, a continuous flow reactor containing immobilized heterotrophic nitrification-aerobic denitrification bacterium Burkholderia sp. YX02 was taken as a model. The microbial community structure in the bioreactor was analyzed by PCR-DCGE, and its correlations with environmental factors such as pH, NH4+ -N, NO2- -N, NO3- -N and COD were simultaneously investigated. The results showed that the microbial community was relatively rich during the early stage of 18 days. The similarity of community structure in different stages was not orderly declining with the operation. In addition, the structural similarity in adjacent stages firstly increased, then decreased, and eventually tended to be stable. Shannon-Wiener index firstly descended significantly, and then ascended with new microbial community emerging at the later stage. UPGMA clustering analysis roughly divided the process into three periods with certain relationship. Principal component analysis showed that during the operation of the bioreactor predominant bacterial community formed steadily and new microbial community dominated by Burkholderia sp. YX02 emerged at the later stage of the operation. Canonical correspondence analysis certificated that the structure of microbial community was most obviously affected by NO2- -N, followed by NO3- -N, NH4+ -N and COD, and pH had the least effect.
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Reactores Biológicos/microbiología , Burkholderia/aislamiento & purificación , Desnitrificación , Consorcios Microbianos , Nitrificación , Procesos HeterotróficosRESUMEN
Demonstrating plausible nonenzymatic polymerization mechanisms for prebiotic monomers represents a fundamental goal in prebiotic chemistry. While a great deal is now known about the potentially prebiotic synthesis of amino acids, our understanding of abiogenic polymerization processes to form polypeptides is less well developed. Here, we show that carbon disulfide (CS2), a component of volcanic emission and sulfide mineral weathering, and a widely used synthetic reagent and solvent, promotes peptide bond formation in modest yields (up to â¼20%) from α-amino acids under mild aqueous conditions. Exposure of a variety of α-amino acids to CS2 initially yields aminoacyl dithiocarbamates, which in turn generate reactive 2-thiono-5-oxazolidone intermediates, the thio analogues of N-carboxyanhydrides. Along with peptides, thiourea and thiohydantoin species are produced. Amino acid stereochemistry was preserved in the formation of peptides. Our findings reveal that CS2 could contribute to peptide bond formation, and possibly other condensation reactions, in abiogenic settings.
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Disulfuro de Carbono/química , Dipéptidos/química , Agua/química , Anaerobiosis , Espectroscopía de Resonancia Magnética con Carbono-13 , Cromatografía Líquida de Alta Presión , Espectroscopía de Protones por Resonancia Magnética , Espectrometría de Masa por Ionización de ElectrosprayRESUMEN
Trisoxazoline 1/Co(ClO(4))(2).6H(2)O catalyzed the 1,3-cycloaddition between nitrones 3 with alkylidene malonates 2 at 0 degrees C to give the isoxazolidines with both high enantioselectivity and high exo selectivity. However, when the temperature was lowered from 0 to -40 degrees C, the same cycloaddition afforded endo isomers as the major products with good to high enantioselectivity. A mechanism is provided.
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Novel chiral iPr-tris(oxazoline)/Cu(ClO4)2 x 6H2O catalyzed coupling of terminal acetylenes and nitrones to afford cis-disubstituted beta-lactams is described; the choice of base proves essential to both the diastereoselectivity and the enantioselectivity.
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Alquinos/química , Cobre/química , Óxidos de Nitrógeno/química , Catálisis , Lactamas/síntesis química , Compuestos Organometálicos/química , Oxazoles/química , EstereoisomerismoAsunto(s)
Dicetopiperazinas/síntesis química , Dipéptidos/química , Biomimética , Catálisis , Péptido SintasasRESUMEN
A systematic analysis using 10 synthetic asimicin stereoisomers revealed that the stereochemistry of the bis-tetrahydrofuran core, including the tetrahydrofuran rings and the adjacent hydroxy functions, had significant effect on its cytotoxicity. Our findings set to rest the highly controversial perception that the stereochemistry of the tetrahydrofuran core has little effect on the activity, which is not true for its cytotoxic effect, and also reinforces the previous conclusion that asimicin is a highly potent anticancer compound.
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Furanos/química , Furanos/toxicidad , Animales , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Furanos/síntesis química , Furanos/farmacología , Humanos , Ratones , Conformación Molecular , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
Ribosomes and nonribosomal peptide synthetases (NRPSs) carry out instructed peptide synthesis through a series of directed intermodular aminoacyl transfer reactions. We recently reported the design of coiled-coil assemblies that could functionally mimic the elementary aminoacyl loading and intermodular aminoacyl transfer steps of NRPSs. These peptides were designed initially to accelerate aminoacyl transfer mainly through catalysis by approximation by closely juxtaposing four active site moieties, two each from adjacent noncovalently associated helical modules. In our designs peptide self-assembly positions a cysteine residue that is used to covalently capture substrates from solution via transthiolesterification (substrate loading step to generate the aminoacyl donor site) adjacent to an aminoacyl acceptor site provided by a covalently tethered amino acid or modeled by the epsilon-amine of an active site lysine. However, through systematic functional analyses of 48 rationally designed peptide sequences, we have now determined that the substrate loading and intermodular aminoacyl transfer steps can be significantly influenced (up to approximately 103-fold) by engineering changes in the active site microenvironment through amino acid substitutions and variations in the inter-residue distances and geometry. Mechanistic studies based on 15N NMR and kinetic analysis further indicate that certain active site constellations furnish an unexpectedly large pK(a) depression (1.5 pH units) of the aminoacyl-acceptor moiety, helping to explain the observed high rates of aminoacyl transfer in those constructs. Taken together, our studies demonstrate the feasibility of engineering efficient de novo peptide sequences possessing active sites and functions reminiscent of those in natural enzymes.
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Materiales Biomiméticos , Péptido Sintasas/química , Péptidos/síntesis química , Ingeniería de Proteínas , Secuencia de Aminoácidos , Aminoácidos , Sitios de Unión , Péptidos/química , Estructura Secundaria de ProteínaRESUMEN
A novel catalyst 2 for Wittig-type and dehalogenation reactions was developed. In the presence of triphenyl phosphite, a wide variety of aldehydes could react with alpha-bromoacetates to afford alpha,beta-unsaturated esters or ketones in high yields with excellent E-stereoselectivity when 1-2 mol % of compound 2 was used. Compound 2 was also an effective catalyst for reductive dehalogenation of alpha-bromocarbonyl compounds. The mechanisms for the above reactions were also proposed.