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α-Melanocyte-stimulating hormone (α-MSH) regulates diverse physiological functions by activating melanocortin receptors (MC-R). However, the role of α-MSH and its possible target receptors in the heart remain completely unknown. Here we investigate whether α-MSH could be involved in pathological cardiac remodeling. We found that α-MSH was highly expressed in the mouse heart with reduced ventricular levels after transverse aortic constriction (TAC). Administration of a stable α-MSH analog protected mice against TAC-induced cardiac hypertrophy and systolic dysfunction. In vitro experiments revealed that MC5-R in cardiomyocytes mediates the anti-hypertrophic signaling of α-MSH. Silencing of MC5-R in cardiomyocytes induced hypertrophy and fibrosis markers in vitro and aggravated TAC-induced cardiac hypertrophy and fibrosis in vivo. Conversely, pharmacological activation of MC5-R improved systolic function and reduced cardiac fibrosis in TAC-operated mice. In conclusion, α-MSH is expressed in the heart and protects against pathological cardiac remodeling by activating MC5-R in cardiomyocytes. These results suggest that analogs of naturally occurring α-MSH, that have been recently approved for clinical use and have agonistic activity at MC5-R, may be of benefit in treating heart failure.
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Remodelação Ventricular , alfa-MSH , Camundongos , Animais , alfa-MSH/farmacologia , Receptores da Corticotropina , Receptores de Melanocortina , Cardiomegalia/genética , FibroseRESUMO
Declines in European bird populations are reported for decades but the direct effect of major anthropogenic pressures on such declines remains unquantified. Causal relationships between pressures and bird population responses are difficult to identify as pressures interact at different spatial scales and responses vary among species. Here, we uncover direct relationships between population time-series of 170 common bird species, monitored at more than 20,000 sites in 28 European countries, over 37 y, and four widespread anthropogenic pressures: agricultural intensification, change in forest cover, urbanisation and temperature change over the last decades. We quantify the influence of each pressure on population time-series and its importance relative to other pressures, and we identify traits of most affected species. We find that agricultural intensification, in particular pesticides and fertiliser use, is the main pressure for most bird population declines, especially for invertebrate feeders. Responses to changes in forest cover, urbanisation and temperature are more species-specific. Specifically, forest cover is associated with a positive effect and growing urbanisation with a negative effect on population dynamics, while temperature change has an effect on the dynamics of a large number of bird populations, the magnitude and direction of which depend on species' thermal preferences. Our results not only confirm the pervasive and strong effects of anthropogenic pressures on common breeding birds, but quantify the relative strength of these effects stressing the urgent need for transformative changes in the way of inhabiting the world in European countries, if bird populations shall have a chance of recovering.
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Agricultura , Florestas , Animais , Fazendas , Europa (Continente) , Dinâmica Populacional , Aves/fisiologia , Biodiversidade , Ecossistema , Conservação dos Recursos NaturaisRESUMO
Plasma apelin levels are reduced in aging and muscle wasting conditions. We aimed to investigate the significance of apelin signaling in cardiac and skeletal muscle responses to physiological stress. Apelin knockout (KO) and wild-type (WT) mice were subjected to high-intensity interval training (HIIT) by treadmill running. The effects of apelin on energy metabolism were studied in primary mouse skeletal muscle myotubes and cardiomyocytes. Apelin increased mitochondrial ATP production and mitochondrial coupling efficiency in myotubes and promoted the expression of mitochondrial genes both in primary myotubes and cardiomyocytes. HIIT induced mild concentric cardiac hypertrophy in WT mice, whereas eccentric growth was observed in the left ventricles of apelin KO mice. HIIT did not affect myofiber size in skeletal muscles of WT mice but decreased the myofiber size in apelin KO mice. The decrease in myofiber size resulted from a fiber type switch toward smaller slow-twitch type I fibers. The increased proportion of slow-twitch type I fibers in apelin KO mice was associated with upregulation of myosin heavy chain slow isoform expression, accompanied with upregulated expression of genes related to fatty acid transport and downregulated expression of genes related to glucose metabolism. Mechanistically, skeletal muscles of apelin KO mice showed defective induction of insulin-like growth factor-1 signaling in response to HIIT. In conclusion, apelin is required for proper skeletal and cardiac muscle adaptation to high-intensity exercise. Promoting apelinergic signaling may have benefits in aging- or disease-related muscle wasting conditions.NEW & NOTEWORTHY Apelin levels decline with age. This study demonstrates that in trained mice, apelin deficiency results in a switch from fast type II myofibers to slow oxidative type I myofibers. This is associated with a concomitant change in gene expression profile toward fatty acid utilization, indicating an aged-muscle phenotype in exercised apelin-deficient mice. These data are of importance in the design of exercise programs for aging individuals and could offer therapeutic target to maintain muscle mass.
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Adaptação Fisiológica , Apelina , Camundongos Knockout , Músculo Esquelético , Condicionamento Físico Animal , Animais , Apelina/metabolismo , Apelina/genética , Camundongos , Condicionamento Físico Animal/fisiologia , Músculo Esquelético/metabolismo , Treinamento Intervalado de Alta Intensidade/métodos , Masculino , Miócitos Cardíacos/metabolismo , Metabolismo Energético , Camundongos Endogâmicos C57BL , Fibras Musculares Esqueléticas/metabolismo , Fibras Musculares de Contração Lenta/metabolismo , Cardiomegalia/metabolismo , Cardiomegalia/genética , Cardiomegalia/fisiopatologia , Cardiomegalia/patologiaRESUMO
Complex dynamic systems displaying interdependency between nitroaldol and boronic ester reactions have been demonstrated. Nitroalkane-1,3-diols, generated by the nitroaldol reaction, were susceptible to ester formation with different boronic acids in aprotic solvents, whereas hydrolysis of the esters occurred in the presence of water. The boronic ester formation led to significant stabilization of the nitroaldol adducts under basic conditions. The use of bifunctional building blocks was furthermore established, allowing for main chain nitroaldol-boronate dynamers as well as complex network dynamers with distinct topologies. The shape and rigidity of the resulting dynamers showed an apparent dependency on the configuration of the boronic acids.
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BACKGROUND: Turning in place is a challenging motor task and is used as a brief assessment test of lower limb function and dynamic balance. This review aims to examine how research of instrumented analysis of turning in place is implemented. In addition to reporting the studied population, we covered acquisition systems, turn detection methods, quantitative parameters, and how these parameters are computed. METHODS: Following the development of a rigorous search strategy, the Web of Science and Scopus were systematically searched for studies involving the use of turning-in-place. From the selected articles, the study population, types of instruments used, turn detection method, and how the turning-in-place characteristics were calculated. RESULTS: Twenty-one papers met the inclusion criteria. The subject groups involved in the reviewed studies included young, middle-aged, and older adults, stroke, multiple sclerosis and Parkinson's disease patients. Inertial measurement units (16 studies) and motion camera systems (5 studies) were employed for gathering measurement data, force platforms were rarely used (2 studies). Two studies used commercial software for turn detection, six studies referenced previously published algorithms, two studies developed a custom detector, and eight studies did not provide any details about the turn detection method. The most frequently used parameters were mean angular velocity (14 cases, 7 studies), turn duration (13 cases, 13 studies), peak angular velocity (8 cases, 8 studies), jerkiness (6 cases, 5 studies) and freezing-of-gait ratios (5 cases, 5 studies). Angular velocities were derived from sensors placed on the lower back (7 cases, 4 studies), trunk (4 cases, 2 studies), and shank (2 cases, 1 study). The rest (9 cases, 8 studies) did not report sensor placement. Calculation of the freezing-of-gait ratio was based on the acceleration of the lower limbs in all cases. Jerkiness computation employed acceleration in the medio-lateral (4 cases) and antero-posterior (1 case) direction. One study did not reported any details about jerkiness computation. CONCLUSION: This review identified the capabilities of turning-in-place assessment in identifying movement differences between the various subject groups. The results, based on data acquired by inertial measurement units across studies, are comparable. A more in-depth analysis of tests developed for gait, which has been adopted in turning-in-place, is needed to examine their validity and accuracy.
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Doença de Parkinson , Acidente Vascular Cerebral , Pessoa de Meia-Idade , Humanos , Idoso , Marcha , Movimento , Perna (Membro)RESUMO
Human tear fluid contains numerous compounds, which are present in highly variable amounts owing to the dynamic and multipurpose functions of tears. A better understanding of the level and sources of variance is essential for determining the functions of the different tear components and the limitations of tear samples as a potential biomarker source. In this study, a quantitative proteomic method was used to analyze variations in the tear protein profiles of healthy volunteers. High day-to-day and inter-eye personal variances were observed in the tear volumes, protein content, and composition of the tear samples. Several normalization and outlier exclusion approaches were evaluated to decrease variances. Despite the intrapersonal variances, statistically significant differences and cluster analysis revealed that proteome profile and immunoglobulin composition of tear fluid present personal characteristics. Using correlation analysis, we could identify several correlating protein clusters, mainly related to the source of the proteins. Our study is the first attempt to achieve more insight into the biochemical background of human tears by statistical evaluation of the experimentally observed dynamic behavior of the tear proteome. As a pilot study for determination of personal protein profiles of the tear fluids of individual patients, it contributes to the application of this noninvasively collectible body fluid in personal medicine.
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Proteoma , Proteômica , Humanos , Proteoma/metabolismo , Proteômica/métodos , Projetos Piloto , Lágrimas/metabolismo , Proteínas do Olho/metabolismo , Controle de QualidadeRESUMO
A reversed-phase high-performance liquid chromatographic (HPLC) method was developed for the simultaneous determination of the potential impurities of dexketoprofen, including the distomer R-ketoprofen. After screening the separation capability of four polysaccharide columns (Lux Amylose-1, Lux Amylose-2, Lux Cellulose-1 and Lux Cellulose-2) in polar organic and in reversed-phase modes, appropriate enantioseparation was observed only on the Lux Amylose-2 column in an acidified acetonitrile/water mixture. A detailed investigation of the mobile phase composition and temperature for enantio- and chemoselectivity showed many unexpected observations. It was observed that both the resolution and the enantiomer elution order can be fine-tuned by varying the temperature and mobile phase composition. Moreover, hysteresis of the retention times and enantioselectivity was also observed in reversed-phase mode using methanol/water mixtures on amylose-type columns. This could indicate that the three-dimensional structure of the amylose column can change by transitioning from a polar organic to a reversed-phase mode, which affects the enantioseparation process. Temperature-dependent enantiomer elution order and rare enthalpic/entropic controlled enantioseparation in the operative temperature range were also observed in reversed-phase mode. To find the best methodological conditions for the determination of dexketoprofen impurities, a full factorial optimization design was performed. Using the optimized parameters (Lux Amylose-2 column with water/acetonitrile/acetic acid 50/50/0.1 (v/v/v) at a 1 mL/min flow rate at 20 °C), baseline separations were achieved between all compounds within 15 min. Our newly developed HPLC method was validated according to the current guidelines, and its application was tested on commercially available pharmaceutical formulations. According to the authors' knowledge, this is the first study to report hysteretic behavior on polysaccharide columns in reversed-phase mode.
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Amilose , Cromatografia de Fase Reversa , Cetoprofeno/análogos & derivados , Trometamina , Amilose/química , Temperatura , Polissacarídeos/química , Celulose/química , Cromatografia Líquida de Alta Pressão/métodos , Água , Acetonitrilas , EstereoisomerismoRESUMO
In systemic sclerosis (SSc), fibrosis of the myocardium along with ongoing autoimmune inflammation can alter the electric function of the cardiac myocytes, which may increase the risk for ventricular arrhythmias and sudden cardiac death. We analyzed the electrocardiographic (ECG) variables describing ventricular repolarization such as QT interval, QT dispersion (QTd), T wave peak-to-end interval (Tpe), and arrhythmogeneity index (AIX) of 26 patients with SSc and 36 healthy controls. Furthermore, echocardiographic and laboratory parameters were examined, with a focus on inflammatory proteins like C-reactive ptotein (CRP), soluble intracellular adhesion molecule-1 (sICAM-1), soluble vascular adhesion molecule-1 (sVCAM-1), and progranulin (PGRN). The CRP, sICAM-1, and sVCAM-1 levels were positively correlated with the length of the QT interval. Although the serum PGRN levels were not increased in the SSc group compared to the controls, in SSc patients, the PGRN levels were positively correlated with the QT interval and the AIX. According to our results, we conclude that there may be a potential association between autoimmune inflammation and the risk for ventricular arrhythmias in patients with SSc. We emphasize that the measurement of laboratory parameters of inflammatory activity including CRP, PGRN, sVCAM-1, and sICAM-1 could be helpful in the prediction of sudden cardiac death in patients with SSc.
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Arritmias Cardíacas , Molécula 1 de Adesão Intercelular , Progranulinas , Escleroderma Sistêmico , Molécula 1 de Adesão de Célula Vascular , Humanos , Escleroderma Sistêmico/sangue , Escleroderma Sistêmico/complicações , Feminino , Masculino , Pessoa de Meia-Idade , Molécula 1 de Adesão de Célula Vascular/sangue , Molécula 1 de Adesão Intercelular/sangue , Arritmias Cardíacas/etiologia , Arritmias Cardíacas/sangue , Progranulinas/sangue , Eletrocardiografia , Adulto , Biomarcadores/sangue , Estudos de Casos e Controles , Idoso , Fatores de Risco , Proteína C-Reativa/metabolismo , Proteína C-Reativa/análiseRESUMO
Acquiring resistance against antiviral drugs is a significant problem in antimicrobial therapy. In order to identify novel antiviral compounds, the antiviral activity of eight plants indigenous to the southern region of Hungary against herpes simplex virus-2 (HSV-2) was investigated. The plant extracts and the plant compound carnosic acid were tested for their effectiveness on both the extracellular and intracellular forms of HSV-2 on Vero and HeLa cells. HSV-2 replication was measured by a direct quantitative PCR (qPCR). Among the tested plant extracts, Salvia rosmarinus (S. rosmarinus) exhibited a 90.46% reduction in HSV-2 replication at the 0.47 µg/mL concentration. Carnosic acid, a major antimicrobial compound found in rosemary, also demonstrated a significant dose-dependent inhibition of both extracellular and intracellular forms of HSV-2. The 90% inhibitory concentration (IC90) of carnosic acid was between 25 and 6.25 µg/mL. Proteomics and high-resolution respirometry showed that carnosic acid suppressed key ATP synthesis pathways such as glycolysis, citrate cycle, and oxidative phosphorylation. Inhibition of oxidative phosphorylation also suppressed HSV-2 replication up to 39.94-fold. These results indicate that the antiviral action of carnosic acid includes the inhibition of ATP generation by suppressing key energy production pathways. Carnosic acid holds promise as a potential novel antiviral agent against HSV-2.
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Abietanos , Trifosfato de Adenosina , Antivirais , Herpesvirus Humano 2 , Extratos Vegetais , Replicação Viral , Abietanos/farmacologia , Replicação Viral/efeitos dos fármacos , Chlorocebus aethiops , Células Vero , Trifosfato de Adenosina/metabolismo , Trifosfato de Adenosina/biossíntese , Humanos , Animais , Herpesvirus Humano 2/efeitos dos fármacos , Herpesvirus Humano 2/fisiologia , Antivirais/farmacologia , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Células HeLaRESUMO
This comprehensive review explores the utilization of chiral stationary phases (CSPs) in the context of single-column simultaneous chiral-achiral high-performance liquid chromatography (HPLC) separation methods. While CSPs have traditionally been pivotal for enantioselective drug analysis, contemporary CSPs often exhibit notable chemoselective properties. Consequently, there is a discernible trend towards the development of methodologies that enable simultaneous enantio- and chemoselective separations utilizing a single CSP-based chromatographic column. This review provides an exhaustive overview of reported HPLC methods in this domain, with a focus on four major CSP types: cyclodextrin-, glycopeptide antibiotic-, protein-, and polysaccharide-based CSPs. This article delves into the diverse applications of CSPs, encompassing various chromatographic modes such as normal phase (NP), reverse phase (RP), and polar organic (PO). This review critically discusses method development, emphasizing the additional chemoselective separation mechanisms of CSPs. It also explores possibilities for method optimization and development, concluding with future perspectives on this evolving field. Despite the inherent challenges in understanding the retention mechanisms involved in chemoselective separations, this review highlights promising trends and anticipates a growing number of simultaneous enantio- and chemoselective methods in pharmaceutical analyses, pharmacokinetic studies, and environmental sample determinations.
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Antibacterianos , Ciclodextrinas , Cromatografia Líquida de Alta Pressão/métodos , Antibacterianos/química , Glicopeptídeos/química , EstereoisomerismoRESUMO
Many enzymes use adaptive frameworks to preorganize substrates, accommodate various structural and electronic demands of intermediates, and accelerate related catalysis. Inspired by biological systems, a Ru-based molecular water oxidation catalyst containing a configurationally labile ligand [2,2':6',2â³-terpyridine]-6,6â³-disulfonate was designed to mimic enzymatic framework, in which the sulfonate coordination is highly flexible and functions as both an electron donor to stabilize high-valent Ru and a proton acceptor to accelerate water dissociation, thus boosting the catalytic water oxidation performance thermodynamically and kinetically. The combination of single-crystal X-ray analysis, various temperature NMR, electrochemical techniques, and DFT calculations was utilized to investigate the fundamental role of the self-adaptive ligand, demonstrating that the on-demand configurational changes give rise to fast catalytic kinetics with a turnover frequency (TOF) over 2000 s-1, which is compared to oxygen-evolving complex (OEC) in natural photosynthesis.
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In the nodules of IRLC legumes, including Medicago truncatula, nitrogen-fixing rhizobia undergo terminal differentiation resulting in elongated and endoreduplicated bacteroids specialized for nitrogen fixation. This irreversible transition of rhizobia is mediated by host produced nodule-specific cysteine-rich (NCR) peptides, of which c. 700 are encoded in the M. truncatula genome but only few of them have been proved to be essential for nitrogen fixation. We carried out the characterization of the nodulation phenotype of three ineffective nitrogen-fixing M. truncatula mutants using confocal and electron microscopy, monitored the expression of defence and senescence-related marker genes, and analysed the bacteroid differentiation with flow cytometry. Genetic mapping combined with microarray- or transcriptome-based cloning was used to identify the impaired genes. Mtsym19 and Mtsym20 mutants are defective in the same peptide NCR-new35 and the lack of NCR343 is responsible for the ineffective symbiosis of NF-FN9363. We found that the expression of NCR-new35 is significantly lower and limited to the transition zone of the nodule compared with other crucial NCRs. The fluorescent protein-tagged version of NCR343 and NCR-new35 localized to the symbiotic compartment. Our discovery added two additional members to the group of NCR genes essential for nitrogen-fixing symbiosis in M. truncatula.
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Medicago truncatula , Rhizobium , Medicago truncatula/genética , Medicago truncatula/metabolismo , Cisteína/metabolismo , Nitrogênio/metabolismo , Peptídeos/metabolismo , Fixação de Nitrogênio , Simbiose , Nódulos Radiculares de Plantas/metabolismoRESUMO
KEY MESSAGE: The bs5 resistance gene against bacterial spot was identified by map-based cloning. The recessive bs5 gene of pepper (Capsicum annuum L.) conditions a non-hypersensitive resistance trait, characterized by a slightly swollen, pale green, photosynthetically active leaf tissue, following Xanthomonas euvesicatoria infection. The isolation of the bs5 gene by map-based cloning revealed that the bs5 protein was shorter by 2 amino acids as compared to the wild type Bs5 protein. The natural 2 amino acid deletion occurred in the cysteine-rich transmembrane domain of the tail-anchored (TA) protein, Ca_CYSTM1. The protein products of the wild type Bs5 and mutant bs5 genes were shown to be located in the cell membrane, indicating an unknown function in this membrane compartment. Successful infection of the Bs5 pepper lines was abolished by the 6 bp deletion in the TM encoding domain of the Ca_CYSTM1 gene in bs5 homozygotes, suggesting, that the resulting resistance might be explained by the lack of entry of the Xanthomonas specific effector molecules into the plant cells.
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Capsicum , Xanthomonas , Capsicum/genética , Capsicum/metabolismo , Alelos , Genes Recessivos , Membrana Celular/metabolismo , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Proteínas de Plantas/genética , Regulação da Expressão Gênica de PlantasRESUMO
Wnt11 regulates early cardiac development and left ventricular compaction in the heart, but it is not known how Wnt11 regulates postnatal cardiac maturation and response to cardiac stress in the adult heart. We studied cell proliferation/maturation in postnatal and adolescent Wnt11 deficient (Wnt11-/-) heart and subjected adult mice with partial (Wnt11+/-) and complete Wnt11 (Wnt11-/-) deficiency to cardiac pressure overload. In addition, we subjected primary cardiomyocytes to recombinant Wnt proteins to study their effect on cardiomyocyte growth. Wnt11 deficiency did not affect cardiomyocyte proliferation or maturation in the postnatal or adolescent heart. However, Wnt11 deficiency led to enlarged heart phenotype that was not accompanied by significant hypertrophy of individual cardiomyocytes. Analysis of stressed adult hearts from wild-type mice showed a progressive decrease in Wnt11 expression in response to pressure overload. When studied in experimental cardiac pressure overload, Wnt11 deficiency did not exacerbate cardiac hypertrophy or remodeling and cardiac function remained identical between the genotypes. When subjecting cardiomyocytes to hypertrophic stimulus, the presence of recombinant Wnt11 together with Wnt5a reduced protein synthesis. In conclusion, Wnt11 deficiency does not affect postnatal cardiomyocyte proliferation but leads to cardiac growth. Interestingly, Wnt11 deficiency alone does not substantially modulate hypertrophic response to pressure overload in vivo. Wnt11 may require cooperation with other noncanonical Wnt proteins to regulate hypertrophic response under stress.
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Coração/crescimento & desenvolvimento , Miócitos Cardíacos/metabolismo , Proteínas Wnt/metabolismo , Animais , Cardiomegalia/metabolismo , Proliferação de Células , Camundongos , Miocárdio , Proteínas Wnt/genéticaRESUMO
Pituitary tumors are rare in chinchillas. This report describes the clinical, gross, histologic, and immunohistochemical characteristics of pituitary tumors in 4 chinchillas. The affected chinchillas were females between 4 and 18 years of age. Clinically, neurologic signs were most commonly reported and included depression, obtundation, seizure, head-pressing, ataxia, and possible blindness. Computed tomography scanning of 2 chinchillas revealed solitary intracranial extra-axial masses in the region of the pituitary gland. Two pituitary tumors were confined to the pars distalis; the other 2 invaded the brain. Based on their microscopic appearances and lack of distant metastases, all 4 tumors were diagnosed as pituitary adenomas. Immunohistochemically, all pituitary adenomas were weakly to strongly positive for growth hormone, most consistent with the diagnosis of somatotropic pituitary adenomas. To the authors' knowledge, this is the first detailed report of the clinical, pathologic, and immunohistochemical features of pituitary tumors in chinchillas.
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Adenoma , Neoplasias Hipofisárias , Doenças dos Roedores , Feminino , Animais , Masculino , Neoplasias Hipofisárias/patologia , Neoplasias Hipofisárias/veterinária , Chinchila , Hipófise/patologia , Adenoma/patologia , Adenoma/veterináriaRESUMO
(1) Background and Goal: Several studies have investigated the association of sleep, diurnal patterns, and circadian rhythms with the presence and with the risk states of mental illnesses such as schizophrenia and bipolar disorder. The goal of our study was to examine actigraphic measures to identify features that can be extracted from them so that a machine learning model can detect premorbid latent liabilities for schizotypy and bipolarity. (2) Methods: Our team developed a small wrist-worn measurement device that collects and identifies actigraphic data based on an accelerometer. The sensors were used by carefully selected healthy participants who were divided into three groups: Control Group (C), Cyclothymia Factor Group (CFG), and Positive Schizotypy Factor Group (PSF). From the data they collected, our team performed data cleaning operations and then used the extracted metrics to generate the feature combinations deemed most effective, along with three machine learning algorithms for categorization. (3) Results: By conducting the training, we were able to identify a set of mildly correlated traits and their order of importance based on the Shapley value that had the greatest impact on the detection of bipolarity and schizotypy according to the logistic regression, Light Gradient Boost, and Random Forest algorithms. (4) Conclusions: These results were successfully compared to the results of other researchers; we had a similar differentiation in features used by others, and successfully developed new ones that might be a good complement for further research. In the future, identifying these traits may help us identify people at risk from mental disorders early in a cost-effective, automated way.
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Transtorno Bipolar , Esquizofrenia , Humanos , Transtorno Bipolar/diagnóstico , Actigrafia/métodos , Esquizofrenia/diagnóstico , Sono , Ritmo CircadianoRESUMO
Altered organokine expression contributes to increased cardiometabolic risk in obesity. Our aim was to evaluate the associations of serum afamin with glucose homeostasis, atherogenic dyslipidemia, and other adipokines in severe obesity to clarify the early metabolic alterations. 106 non-diabetic obese (NDO) subjects and 62 obese patients with type 2 diabetes matched for age, gender, and body mass index (BMI) were enrolled in this study. We compared their data with 49 healthy, lean controls. Serum afamin and retinol-binding protein 4 (RBP4), as well as plasma plasminogen activator inhibitor-1 (PAI-1), were measured with ELISA, and lipoprotein subfractions were analyzed using Lipoprint gel electrophoresis. Afamin and PAI-1 found to be significantly higher in the NDO and T2M group (p < 0.001 and p < 0.001, respectively) than in the controls. In contrast, RBP4 was unexpectedly lower in the NDO and T2DM group compared to controls (p < 0.001). Afamin showed negative correlations with mean LDL size and RBP4, but positive correlations with anthropometric, glucose/lipid parameters, and PAI-1 in both the overall patients and the in NDO + T2DM groups. BMI, glucose, intermediate HDL, and small HDL were predictors of afamin. Afamin may serve as a biomarker for the severity of cardiometabolic disturbances in obesity. The complexity of organokine patterns in NDO subjects draws attention to the diverse spectrum of obesity-related comorbidities.
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Doenças Cardiovasculares , Diabetes Mellitus Tipo 2 , Resistência à Insulina , Humanos , Inibidor 1 de Ativador de Plasminogênio/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Obesidade/metabolismo , Índice de Massa Corporal , Glucose , Proteínas Plasmáticas de Ligação ao RetinolRESUMO
The interaction between human serum albumin (HSA) and apremilast (APR), a novel antipsoriatic drug, was characterized by multimodal analytical techniques including high-performance liquid chromatography (HPLC), fluorescence spectroscopy and molecular docking for the first time. Using an HSA chiral stationary phase, the APR enantiomers were well separated, indicating enantioselective binding between the protein and the analytes. The influence of chromatographic parameters-type and concentration of the organic modifier, buffer type, pH, ionic strength of the mobile phase, flow rate and column temperature-on the chromatographic responses (retention factor and selectivity) was analyzed in detail. The results revealed that the eutomer S-APR bound to the protein to a greater extent than the antipode. The classical van 't Hoff method was applied for thermodynamic analysis, which indicated that the enantioseparation was enthalpy-controlled. The stability constants of the protein-enantiomer complexes, determined by fluorescence spectroscopy, were in accordance with the elution order observed in HPLC (KR-APR-HSA = 6.45 × 103 M-1, KS-APR-HSA = 1.04 × 104 M-1), showing that, indeed, the later-eluting S-APR displayed a stronger binding with HSA. Molecular docking was applied to study and analyze the interactions between HSA and the APR enantiomers at the atomic level. It was revealed that the most favored APR binding occurred at the border between domains I and II of HSA, and secondary interactions were responsible for the different binding strengths of the enantiomers.
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Albumina Sérica Humana , Albumina Sérica , Humanos , Albumina Sérica Humana/metabolismo , Simulação de Acoplamento Molecular , Albumina Sérica/metabolismo , Estereoisomerismo , Cromatografia Líquida de Alta Pressão/métodos , Termodinâmica , Ligação Proteica , Sítios de Ligação , Espectrometria de FluorescênciaRESUMO
Four diastereomers of 16-azidomethyl substituted 3-O-benzyl estradiol (1-4) and their two estrone analogs (16AABE and 16BABE) were tested for their antiproliferative properties against human gynecological cancer cell lines. The estrones were selected for additional experiments based on their outstanding cell growth-inhibiting activities. Both compounds increased hypodiploid populations of breast cancer cells, and 16AABE elicited cell cycle disturbance as evidenced by flow cytometry. The two analogs substantially increased the rate of tubulin polymerization in vitro. 16AABE and 16BABE inhibited breast cancer cells' migration and invasive ability, as evidenced by wound healing and Boyden chamber assays. Since both estrone analogs exerted remarkable estrogenic activities, as documented by a luciferase reporter gene assay, they can be considered as promising drug candidates for hormone-independent malignancies.
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Neoplasias da Mama , Estrona , Humanos , Feminino , Estrona/farmacologia , Estradiol , Aneuploidia , Bioensaio , Neoplasias da Mama/tratamento farmacológicoRESUMO
Mechanistic modeling is useful for predicting and modulating selectivity even in early chromatographic method development. This approach is also in accordance with current analytical quality using design principles and is highly welcomed by the authorities. The aim of this study was to investigate the separation behavior of two different types of chiral stationary phases (CSPs) for the separation of ezetimibe and its related substances using the mechanistic retention modeling approach offered by the Drylab software (version 4.5) package. Based on the obtained results, both CSPs presented with chemoselectivity towards the impurities of ezetimibe. The cyclodextrin-based CSP displayed a higher separation capacity and was able to separate seven related substances from the active pharmaceutical ingredient, while the cellulose-based column enabled the baseline resolution of six impurities from ezetimibe. Generally, the accuracy of predicted retention times was lower for the polysaccharide CSP, which could indicate the presence of additional secondary interactions between the analytes and the CSP. It was also demonstrated that the combination of mechanistic modeling and an experimental design approach can be applied to method development on CSPs in reverse-phase mode. The applicability of the methods was tested on spiked artificial placebo samples, while intraday and long-term (2 years) method repeatability was also challenged through comparing the obtained retention times and resolution values. The results indicated the excellent robustness of the selected setpoints. Overall, our findings indicate that the chiral columns could offer orthogonal selectivity to traditional reverse-phase columns for the separation of structurally similar compounds.