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1.
Sci Rep ; 13(1): 17164, 2023 10 11.
Artículo en Inglés | MEDLINE | ID: mdl-37821527

RESUMEN

Aimed to explore the relationships between infection localization, bacterial species, and procalcitonin (PCT) and C-reactive protein (CRP) levels in urinary tract infections (UTIs). A retrospective study included 314 UTI hospitalized patients divided into two groups (268 with lower UTI, 46 with upper UTI) in a tertiary care hospital. PCT and CRP were performed. Bacterial isolates were identified using standard microbiological techniques, and statistical analyses were performed to assess associations between infection localization, bacterial species, PCT, and CRP levels. Age and gender showed no significant differences between the lower and upper UTIs. Escherichia coli dominated as the leading UTI pathogen. A positive correlation (r = 0.646, P < 0.001) between PCT and CRP levels was found. The subgroup with ureteritis in the upper UTI category exhibited the highest PCT and CRP levels. PCT and CRP exhibited favorable diagnostic potential in predicting upper UTIs, with AUCs of 0.644 and 0.629, respectively. The optimal cutoff values were 0.21 ng/mL for PCT and 60.77 mg/L for CRP. Sensitivities were 69.03% and 77.99%, while specificities were 56.52% and 47.83%, respectively. E. coli emerged as the predominant bacterium in UTIs. PCT and CRP demonstrated moderate diagnostic efficacy in distinguishing between upper and lower UTIs. Notably, PCT and CRP exhibited enhanced utility in identifying ureteritis.


Asunto(s)
Polipéptido alfa Relacionado con Calcitonina , Infecciones Urinarias , Humanos , Proteína C-Reactiva/análisis , Biomarcadores/análisis , Calcitonina , Estudios Retrospectivos , Escherichia coli , Péptido Relacionado con Gen de Calcitonina , Precursores de Proteínas/análisis , Infecciones Urinarias/diagnóstico
2.
Biomolecules ; 12(12)2022 11 29.
Artículo en Inglés | MEDLINE | ID: mdl-36551209

RESUMEN

Alterations to the intestinal barrier may be involved in the pathogenesis of various chronic diseases. The diagnosis of mucosal barrier disruption has become a new therapeutic target for disease prevention. The aim of this study was to determine whether various patient demographic and biometric data, often not included in diagnostic analyses, may affect calprotectin, zonulin, and sIgA biomarker values. Stool markers' levels in 160 samples were measured colorimetrically. The analysis of twenty key bacteria (15 genera and 5 species) was carried out on the basis of diagnostic tests, including cultures and molecular tests. The concentrations of selected markers were within reference ranges for most patients. The sIgA level was significantly lower in participants declaring probiotics supplementation (p = 0.0464). We did not observe differences in gastrointestinal discomfort in participants. We found significant differences in the sIgA level between the 29-55 years and >55 years age-related intervals groups (p = 0.0191), together with a significant decreasing trend (p = 0.0337) in age-dependent sIgA concentration. We observed complex interdependencies and relationships between their microbiota and the analyzed biomarkers. For correct clinical application, standardized values of calprotectin and sIgA should be determined, especially in elderly patients. We observed a correlation between the composition of the gut community and biomarker levels, although it requires further in-depth analysis.


Asunto(s)
Heces , Microbioma Gastrointestinal , Haptoglobinas , Inmunoglobulina A Secretora , Complejo de Antígeno L1 de Leucocito , Probióticos , Precursores de Proteínas , Adulto , Anciano , Humanos , Biomarcadores/análisis , Biometría , Inmunoglobulina A Secretora/análisis , Inmunoglobulina A Secretora/metabolismo , Probióticos/administración & dosificación , Haptoglobinas/análisis , Precursores de Proteínas/análisis , Masculino , Femenino , Adolescente , Adulto Joven , Persona de Mediana Edad
3.
Bioanalysis ; 14(18): 1229-1239, 2022 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-36378599

RESUMEN

Background & Aim: Oxyntomodulin (Oxm) is a proglucagon-derived peptide agonist of both the GLP-1 and glucagon receptors and is a key regulator of gastric acid secretion and energy expenditure. Differential processing from proglucagon hinders assay immunoassay selectivity. Method & results: Antibody engineering was used to develop a sandwich immunoassay that selectively measures endogenous Oxm. The pre- and postprandial levels of Oxm from 19 healthy individuals over the course of 2 h were measured. Postprandial increases in Oxm occurred within minutes and levels significantly correlated with those obtained using previously published mass spectrometry assays. Conclusion: This sandwich immunoassay is appropriately sensitive and selective and is also amenable to high-throughput application for the reliable determination of endogenous levels of intact Oxm from human samples.


Asunto(s)
Anticuerpos Monoclonales , Oxintomodulina , Humanos , Proglucagón , Glucagón , Precursores de Proteínas/análisis , Péptido 1 Similar al Glucagón , Inmunoensayo
4.
Pediatr Crit Care Med ; 23(12): 980-989, 2022 12 01.
Artículo en Inglés | MEDLINE | ID: mdl-36239515

RESUMEN

OBJECTIVES: Current sepsis guidelines do not provide good risk stratification of subgroups in whom prompt IV antibiotics and fluid resuscitation might of benefit. We evaluated the utility of mid-regional pro-adrenomedullin (MR-proADM) in identification of patient subgroups at risk of requiring PICU or high-dependency unit (HDU) admission or fluid resuscitation. DESIGN: Secondary, nonprespecified analysis of prospectively collected dataset. SETTING: Pediatric Emergency Department in a United Kingdom tertiary center. PATIENTS: Children less than 16 years old presenting with fever and clinical indication for venous blood sampling ( n = 1,183). INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Primary outcome measures were PICU/HDU admission or administration of fluid resuscitation, with a secondary outcome of definite or probable bacterial infection. Biomarkers were measured on stored plasma samples and children phenotyped into bacterial and viral groups using a previously published algorithm. Of the 1,183 cases, 146 children (12.3%) required fluids, 48 (4.1%) were admitted to the PICU/HDU, and 244 (20.6%) had definite or probable bacterial infection. Area under the receiver operating characteristic (AUC) was used to assess performance. MR-proADM better predicted fluid resuscitation (AUC, 0.73; 95% CI, 0.67-0.78), than both procalcitonin (AUC, 0.65; 95% CI, 0.59-0.71) and Pediatric Early Warning Score (PEWS: AUC, 0.62; 95% CI, 0.56-0.67). PEWS alone showed good accuracy for PICU/HDU admission 0.83 (0.78-0.89). Patient subgroups with high MR-proADM (≥ 0.7 nmol/L) and high procalcitonin (≥ 0.5 ng/mL) had increased association with PICU/HDU admission, fluid resuscitation, and bacterial infection compared with subgroups with low MR-proADM (< 0.7 nmol/L). For children with procalcitonin less than 0.5 ng/mL, high MR-proADM improved stratification for fluid resuscitation only. CONCLUSIONS: High MR-proADM and high procalcitonin were associated with increased likelihood of subsequent disease progression. Incorporating MR-proADM into clinical risk stratification may be useful in clinician decision-making regarding initiation of IV antibiotics, fluid resuscitation, and escalation to PICU/HDU admission.


Asunto(s)
Infecciones Bacterianas , Puntuación de Alerta Temprana , Humanos , Niño , Adolescente , Adrenomedulina/análisis , Polipéptido alfa Relacionado con Calcitonina , Estudios de Cohortes , Precursores de Proteínas/análisis , Servicio de Urgencia en Hospital , Biomarcadores , Fiebre/diagnóstico , Fiebre/etiología , Fiebre/tratamiento farmacológico , Infecciones Bacterianas/tratamiento farmacológico , Reino Unido , Medición de Riesgo , Antibacterianos/uso terapéutico , Pronóstico
5.
Eur Rev Med Pharmacol Sci ; 25(13): 4639-4643, 2021 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-34286505

RESUMEN

OBJECTIVE: Acute respiratory distress syndrome (ARDS) is characterized by quantitative and qualitative changes in surfactant composition, leading to surfactant dysregulation with alveolar collapse and acute respiratory hypoxic failure. Recently, surfactant has been hypothesized to play a relevant role in COVID-19, representing a strong defender against SARS-CoV-2 infection. The aim of our work was the study of immunohistochemical surfactant expression in the lungs of patients died following SARS-CoV-2 ARDS, in order to shed light on a possible therapeutic surfactant administration. PATIENTS AND METHODS: We investigated four patients who died due to ARDS following SARS-COV-2 infection and four patients submitted to lung biopsy, in the absence of SARS-CoV-2 infection. In all 8 cases, lung specimens were immunostained with anti-surfactant protein A (SP-A) and B (SP-B). RESULTS: In control subjects, reactivity for SP-B was restricted to type II alveolar cells. Immunostaining for SP-A was observed on the surface of alveolar spaces. In the COVID-19 positive lungs, immunoreactivity for SP-B was similar to that observed in control lungs; SP-A was strongly expressed along the alveolar wall. Moreover, dense aggregates of SP-A positive material were observed in the alveolar spaces. CONCLUSIONS: Our immunohistochemical data show the dysregulation of surfactant production in COVID-19 patients, particularly regarding SP-A expression. The increased presence of SP-A in condensed masses inside alveolar spaces could invalidate the therapeutic efficacy of the treatment with exogenous surfactant.


Asunto(s)
COVID-19/metabolismo , Inmunohistoquímica , Precursores de Proteínas/análisis , Proteína A Asociada a Surfactante Pulmonar/análisis , Proteínas Asociadas a Surfactante Pulmonar/análisis , COVID-19/diagnóstico por imagen , Humanos , Precursores de Proteínas/genética , Precursores de Proteínas/metabolismo , Alveolos Pulmonares/diagnóstico por imagen , Alveolos Pulmonares/metabolismo , Proteína A Asociada a Surfactante Pulmonar/genética , Proteína A Asociada a Surfactante Pulmonar/metabolismo , Proteínas Asociadas a Surfactante Pulmonar/genética , Proteínas Asociadas a Surfactante Pulmonar/metabolismo , Estudios Retrospectivos , SARS-CoV-2/aislamiento & purificación , SARS-CoV-2/metabolismo
6.
Anal Biochem ; 629: 114311, 2021 09 15.
Artículo en Inglés | MEDLINE | ID: mdl-34302800

RESUMEN

The choriogenin H - EGFP transgenic medaka (Oryzias melastigma) has been used to test estrogenic substances and quantify estrogenic activity into 17ß-estradiol (E2) equivalency (EEQ). The method uses 8 eleutheroembryos in 2 ml solution per well and 3 wells per treatment in 24-well plates at 26 ± 1 °C for 24 ± 2 h, with subsequent measurements of induced GFP signal intensity. EEQ measurements are calculated using a E2 probit regression model with a coefficient of determination (R2) > 0.90. The selectivity was confirmed evaluating 27 known estrogenic and 5 known non-estrogenic compounds. Limit of quantitation (LOQ), recovery rate and bias were calculated to be 1 ng/ml EEQ, 104% and 4% respectively. Robustness analysis revealed exposure temperature is a sensitive parameter that should be kept at 26 ± 1 °C. The repeatability of intra- and inter-laboratories achieved CV < 30% for most tested food and cosmetics samples. The lot-lot stability was confirmed by the stable EEQ qualitative control (QC, 1 ng/mL E2) and calibration curve results. The stability of standard reagents, samples and sample extracts was also investigated. These data demonstrated this method to be an accurate indicator of estrogenic activity for both chemicals and extracts.


Asunto(s)
Animales Modificados Genéticamente/metabolismo , Proteínas del Huevo/análisis , Estradiol/química , Oryzias/metabolismo , Precursores de Proteínas/análisis , Animales , Animales Modificados Genéticamente/embriología , Técnicas Biosensibles , Extractos Celulares/química , Estradiol/metabolismo , Límite de Detección , Oryzias/embriología , Análisis de Regresión
7.
Respir Res ; 22(1): 148, 2021 May 13.
Artículo en Inglés | MEDLINE | ID: mdl-33985491

RESUMEN

BACKGROUND: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) disease (COVID-19) has been linked to thrombotic complications and endothelial dysfunction. We assessed the prognostic implications of endothelial activation through measurement of endothelin-I precursor peptide (proET-1), the stable precursor protein of Endothelin-1, in a well-defined cohort of patients hospitalized with COVID-19. METHODS: We measured proET-1 in 74 consecutively admitted adult patients with confirmed COVID-19 and compared its prognostic accuracy to that of patients with community-acquired pneumonia (n = 876) and viral bronchitis (n = 371) from a previous study by means of logistic regression analysis. The primary endpoint was all-cause 30-day mortality. RESULTS: Overall, median admission proET-1 levels were lower in COVID-19 patients compared to those with pneumonia and exacerbated bronchitis, respectively (57.0 pmol/l vs. 113.0 pmol/l vs. 96.0 pmol/l, p < 0.01). Although COVID-19 non-survivors had 1.5-fold higher admission proET-1 levels compared to survivors (81.8 pmol/l [IQR: 76 to 118] vs. 53.6 [IQR: 37 to 69]), no significant association of proET-1 levels and mortality was found in a regression model adjusted for age, gender, creatinine level, diastolic blood pressure as well as cancer and coronary artery disease (adjusted OR 0.1, 95% CI 0.0009 to 14.7). In patients with pneumonia (adjusted OR 25.4, 95% CI 5.1 to 127.4) and exacerbated bronchitis (adjusted OR 120.1, 95% CI 1.9 to 7499) we found significant associations of proET-1 and mortality. CONCLUSIONS: Compared to other types of pulmonary infection, COVID-19 shows only a mild activation of the endothelium as assessed through measurement of proET-1. Therefore, the high mortality associated with COVID-19 may not be attributed to endothelial dysfunction by the surrogate marker proET-1.


Asunto(s)
COVID-19/mortalidad , COVID-19/fisiopatología , Endotelina-1/análisis , Endotelio Vascular/fisiopatología , Precursores de Proteínas/análisis , Factores de Edad , Anciano , Anciano de 80 o más Años , Biomarcadores/análisis , Presión Sanguínea , Estudios de Cohortes , Creatinina/sangre , Determinación de Punto Final , Femenino , Humanos , Masculino , Persona de Mediana Edad , Pronóstico , Estudios Prospectivos , Reproducibilidad de los Resultados , Factores de Riesgo , Factores Sexuales , Análisis de Supervivencia
8.
J Am Soc Mass Spectrom ; 32(6): 1352-1360, 2021 Jun 02.
Artículo en Inglés | MEDLINE | ID: mdl-33605729

RESUMEN

Crustacean hyperglycemic hormones (CHHs) are a family of neuropeptides that were discovered in multiple tissues in crustaceans, but the function of most isoforms remains unclear. Functional discovery often requires comprehensive qualitative profiling and quantitative analysis. The conventional enzymatic digestion method has several limitations, such as missing post-translational modification (PTM) information, homology interference, and incomplete sequence coverage. Herein, by using a targeted top-down method, facilitated by higher sensitivity instruments and hybrid fragmentation modes, we achieved the characterization of two CHH isoforms from the sinus glands (SG-CHH) and the pericardial organs (PO-CHH) from the Atlantic blue crab, Callinectes sapidus, with improved sequence coverage compared to earlier studies. In this study, both label-free and isotopic labeling approaches were adopted to monitor the response of CHHs and CHH precursor-related peptide (CPRP) under low pH stress. The identical trends of CPRP and CHH expression indicated that CPRP could serve as an ideal probe in tracking the CHH expression level changes, which would greatly simplify the quantitative analysis of large peptides. Furthermore, the distinct patterns of changes in the expression of CHHs in the SG and the PO suggested their tissue-specific functions in the regulation of low pH stress. Ion mobility-mass spectrometry (IM-MS) was also employed in this study to provide conformation analysis of both CHHs and CPRPs from different tissues.


Asunto(s)
Proteínas de Artrópodos/análisis , Braquiuros/química , Braquiuros/fisiología , Hormonas de Invertebrados/análisis , Espectrometría de Masas/métodos , Proteínas del Tejido Nervioso/análisis , Precursores de Proteínas/análisis , Animales , Proteínas de Artrópodos/química , Proteínas de Artrópodos/metabolismo , Concentración de Iones de Hidrógeno , Hormonas de Invertebrados/química , Hormonas de Invertebrados/metabolismo , Espectrometría de Movilidad Iónica , Proteínas del Tejido Nervioso/química , Proteínas del Tejido Nervioso/metabolismo , Especificidad de Órganos , Péptidos/análisis , Péptidos/metabolismo , Isoformas de Proteínas/análisis , Isoformas de Proteínas/metabolismo , Precursores de Proteínas/química , Precursores de Proteínas/metabolismo , Proteómica/métodos , Estrés Fisiológico
9.
J Comp Neurol ; 529(3): 635-654, 2021 02.
Artículo en Inglés | MEDLINE | ID: mdl-32602558

RESUMEN

Sensory information is transmitted from peripheral nerves, through the spinal cord, and up to the brain. Sensory information may be modulated by projections from the brain to the spinal cord, but the neural substrates for top-down sensory control are incompletely understood. We identified a novel population of inhibitory neurons in the mouse brainstem, distinguished by their expression of prodynorphin, which we named LJA5. Here, we identify a similar group of Pdyn+ neurons in the human brainstem, and we define the efferent and afferent projection patterns of LJA5 neurons in mouse. Using specific genetic tools, we selectively traced the projections of the Pdyn-expressing LJA5 neurons through the brain and spinal cord. Terminal fields were densest in the lateral and ventrolateral periaqueductal gray (PAG), lateral parabrachial nucleus (LPB), caudal pressor area, and lamina I of the spinal trigeminal nucleus and all levels of the spinal cord. We then labeled cell types in the PAG, LPB, medulla, and spinal cord to better define the specific targets of LJA5 boutons. LJA5 neurons send the only known inhibitory descending projection specifically to lamina I of the spinal cord, which transmits afferent pain, temperature, and itch information up to the brain. Using retrograde tracing, we found LJA5 neurons receive inputs from sensory and stress areas such as somatosensory/insular cortex, preoptic area, paraventricular nucleus, dorsomedial nucleus and lateral hypothalamus, PAG, and LPB. This pattern of inputs and outputs suggest LJA5 neurons are uniquely positioned to be activated by sensation and stress, and in turn, inhibit pain and itch.


Asunto(s)
Tronco Encefálico/química , Tronco Encefálico/metabolismo , Encefalinas/análisis , Encefalinas/metabolismo , Neuronas/química , Neuronas/metabolismo , Precursores de Proteínas/análisis , Precursores de Proteínas/metabolismo , Animales , Tronco Encefálico/citología , Humanos , Recién Nacido , Ratones , Ratones Transgénicos
10.
J Comp Neurol ; 529(4): 657-693, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-32621762

RESUMEN

The parabrachial nucleus (PB) is a complex structure located at the junction of the midbrain and hindbrain. Its neurons have diverse genetic profiles and influence a variety of homeostatic functions. While its cytoarchitecture and overall efferent projections are known, we lack comprehensive information on the projection patterns of specific neuronal subtypes in the PB. In this study, we compared the projection patterns of glutamatergic neurons here with a subpopulation expressing the transcription factor Foxp2 and a further subpopulation expressing the neuropeptide Pdyn. To do this, we injected an AAV into the PB region to deliver a Cre-dependent anterograde tracer (synaptophysin-mCherry) in three different strains of Cre-driver mice. We then analyzed 147 neuroanatomical regions for labeled boutons in every brain (n = 11). Overall, glutamatergic neurons in the PB region project to a wide variety of sites in the cerebral cortex, basal forebrain, bed nucleus of the stria terminalis, amygdala, diencephalon, and brainstem. Foxp2 and Pdyn subpopulations project heavily to the hypothalamus, but not to the cortex, basal forebrain, or amygdala. Among the few differences between Foxp2 and Pdyn cases was a notable lack of Pdyn projections to the ventromedial hypothalamic nucleus. Our results indicate that genetic identity determines connectivity (and therefore, function), providing a framework for mapping all PB output projections based on the genetic identity of its neurons. Using genetic markers to systematically classify PB neurons and their efferent projections will enhance the translation of research findings from experimental animals to humans.


Asunto(s)
Encefalinas/biosíntesis , Factores de Transcripción Forkhead/biosíntesis , Núcleos Parabraquiales/metabolismo , Precursores de Proteínas/biosíntesis , Proteínas Represoras/biosíntesis , Proteína 2 de Transporte Vesicular de Glutamato/biosíntesis , Animales , Tronco Encefálico/química , Tronco Encefálico/metabolismo , Corteza Cerebral/química , Corteza Cerebral/metabolismo , Vías Eferentes/química , Vías Eferentes/metabolismo , Encefalinas/análisis , Encefalinas/genética , Femenino , Factores de Transcripción Forkhead/análisis , Factores de Transcripción Forkhead/genética , Hipotálamo/química , Hipotálamo/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Núcleos Parabraquiales/química , Precursores de Proteínas/análisis , Precursores de Proteínas/genética , Proteínas Represoras/análisis , Proteínas Represoras/genética , Tálamo/química , Tálamo/metabolismo , Proteína 2 de Transporte Vesicular de Glutamato/análisis , Proteína 2 de Transporte Vesicular de Glutamato/genética
11.
Ann Endocrinol (Paris) ; 82(1): 1-7, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33278377

RESUMEN

BACKGROUND: This study aimed to investigate the relationship between lipopolysaccharide (LPS) and zonulin levels and also to show the effect of acute hyperglycemic stress induced by oral glucose tolerance testing (OGTT) on zonulin levels in pre-diabetic patients. METHODS: Four groups were constituted according to the criteria of the American Diabetes Association (ADA), based on OGTT results: control group (n:40); prediabetic group (n:56), divided into two subgroups: impaired fasting glucose group (IFG) (n:36), and impaired glucose tolerance (IGT) + IFG group (n:20) and type-2 diabetes mellitus (T2DM) group (n:45). RESULTS: Zonulin and LPS did not significantly differ between the prediabetes and control groups, but were significantly higher in the T2DM group compared to both the prediabetic and the control group (P<0.001). After OGTT, zonulin and LPS were significantly higher in the prediabetes group compared to the control group (P<0.01 and P<0.05, respectively), and significantly lower in the IFG and IFG+IGT groups compared to the T2DM group (P<0.001, P<0.001 and P<0.001, P<0.001, respectively). A positive correlation was detected between fasting zonulin and 2-hour zonulin (r=0.727, P<0.001) and between fasting LPS (r=0.555, P<0.001) and 2-hour LPS (r=0.567, P<0.001) in the prediabetic group. Increased zonulin and LPS levels and the positive correlation between these levels during the prediabetic period although non significant suggests onset of intestinal permeability. CONCLUSIONS: During acute hyperglycemia in prediabetic patients, up-regulation of zonulin and LPS may affect intestinal function. The intestines may play a key role in up-regulation of glucose and the pathogenesis of diabetes.


Asunto(s)
Estado Prediabético/sangre , Precursores de Proteínas/sangre , Adulto , Glucemia/análisis , Glucemia/metabolismo , Estudios de Casos y Controles , Diabetes Mellitus Tipo 2/sangre , Ayuno/sangre , Femenino , Intolerancia a la Glucosa/sangre , Prueba de Tolerancia a la Glucosa , Haptoglobinas/análisis , Haptoglobinas/metabolismo , Humanos , Hiperglucemia/sangre , Lipopolisacáridos/análisis , Lipopolisacáridos/sangre , Masculino , Persona de Mediana Edad , Precursores de Proteínas/análisis , Turquía
12.
J Am Soc Mass Spectrom ; 32(2): 611-614, 2021 Feb 03.
Artículo en Inglés | MEDLINE | ID: mdl-33382608

RESUMEN

This initial study evaluates vacuum matrix-assisted ionization (vMAI) mass spectrometry (MS) for identification and determination of tryptic peptides from the biomarker protein progastrin releasing peptide (ProGRP). Similar peptides and charge states were observed as in liquid chromatography (LC) electrospray ionization (ESI) MS. The prolonged ion duration in vMAI with similar charge states as in ESI was advantageous for determining the MS/MS fragmentation conditions compared to MAI. It is assumed that the vacuum ionization conditions lower the detection limits of the experiment. This may be the reason vMAI combined with high resolution MS enabled detection of tryptic peptides from more digested proteins than MAI selected reaction monitoring MS. Additionally, MAI ion mobility spectrometry MS (MAI-IMS-MS) was evaluated for differentiation of intact protein isoforms, successfully enabling differentiation of the isoforms by drift time selection. Examples are both shown for model proteins bovine serum albumin, cytochrome C, and lysozyme and the clinically relevant small cell lung cancer protein biomarker ProGRP, which exists in three isoforms. Coupling with the vacuum ionization conditions using a dedicated vacuum-probe source MAI enables information to be extracted readily as with conventional approaches, just faster.


Asunto(s)
Biomarcadores de Tumor/análisis , Precursores de Proteínas/análisis , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Espectrometría de Masas en Tándem/métodos , Humanos , Neoplasias Pulmonares/metabolismo , Fragmentos de Péptidos/análisis , Isoformas de Proteínas/análisis , Carcinoma Pulmonar de Células Pequeñas/metabolismo , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/instrumentación , Espectrometría de Masas en Tándem/instrumentación
15.
Viruses ; 12(9)2020 08 26.
Artículo en Inglés | MEDLINE | ID: mdl-32859114

RESUMEN

Chronic hepatitis B virus (HBV) infection is a major risk factor for the development of hepatocellular carcinoma (HCC), the leading cause of cancer-related death worldwide. Despite progress in the prevention and therapy of HCC, high incidence and recurrence rates of HCC remain big threats, resulting in poor patient survival. Effective biomarkers and targets of HCC are therefore urgently needed for better management and to improve patient outcomes. Pre-S mutants have been well demonstrated as HBV oncoproteins that play important roles in HCC development through activation of multiple oncogenic signal pathways in hepatocytes, in vitro and in vivo. The presence of pre-S mutants in patients with chronic HBV infection and HBV-related HCC has been associated with a significantly higher risk of HCC development and recurrence after curative surgical resection, respectively. In this review, we summarize the roles of pre-S mutants as biomarkers for predicting HBV-related HCC development and recurrence, and highlight the pre-S mutants-activated oncogenic signal pathways as potential targets for preventing HBV-related HCC development.


Asunto(s)
Biomarcadores/análisis , Carcinoma Hepatocelular/diagnóstico , Antígenos de Superficie de la Hepatitis B/análisis , Hepatitis B Crónica/complicaciones , Hepatitis B Crónica/virología , Neoplasias Hepáticas/diagnóstico , Precursores de Proteínas/análisis , Antivirales/uso terapéutico , Biomarcadores/sangre , Carcinoma Hepatocelular/etiología , Carcinoma Hepatocelular/prevención & control , Carcinoma Hepatocelular/cirugía , Antígenos de Superficie de la Hepatitis B/sangre , Antígenos de Superficie de la Hepatitis B/genética , Virus de la Hepatitis B/genética , Hepatitis B Crónica/tratamiento farmacológico , Hepatitis B Crónica/metabolismo , Humanos , Cirrosis Hepática/diagnóstico , Cirrosis Hepática/etiología , Neoplasias Hepáticas/etiología , Neoplasias Hepáticas/prevención & control , Neoplasias Hepáticas/cirugía , Proteínas Mutantes/sangre , Mutación , Recurrencia Local de Neoplasia , Precursores de Proteínas/sangre , Precursores de Proteínas/genética , Factores de Riesgo , Transducción de Señal
16.
Nutrients ; 12(8)2020 Aug 05.
Artículo en Inglés | MEDLINE | ID: mdl-32764250

RESUMEN

(1) Background: Psoriatic Arthritis (PsA) is a painful disease of the joints and spine. Recent reports observed distinct enteric dysbiosis in PsA; intake of probiotic strains is considered to ameliorate enteric dysbiosis. If probiotics are effective in PsA is elusive. (2) Methods: In this pilot open-label study we enrolled 10 PsA patients with low to medium disease activity who received probiotics for 12 weeks. Analysis of faecal zonulin, α1-antitrypsin and calprotectin, as well as peripheral immune phenotyping was performed at baseline, after 12 weeks and 12 weeks after termination of probiotic intake. (3) Results: All patients showed increased levels of the enteric permeability marker zonulin which correlated with the frequency of peripheral Th17 cells. Calprotectin, a marker for intestinal inflammation was elevated in 6 out of 10 patients. Probiotic intake resulted in a reduction of disease activity and gut permeability. These effects, however, were not sustained beyond termination of probiotic intake. (4) Conclusions: PsA patients suffer from enhanced enteric permeability and inflammation. Probiotics may ameliorate disease activity in PsA by targeting these alterations.


Asunto(s)
Artritis Psoriásica/tratamiento farmacológico , Disbiosis/tratamiento farmacológico , Inflamación/tratamiento farmacológico , Mucosa Intestinal/efectos de los fármacos , Probióticos/administración & dosificación , Anciano , Heces/química , Femenino , Microbioma Gastrointestinal/efectos de los fármacos , Haptoglobinas/análisis , Humanos , Mucosa Intestinal/metabolismo , Complejo de Antígeno L1 de Leucocito/análisis , Masculino , Persona de Mediana Edad , Permeabilidad/efectos de los fármacos , Proyectos Piloto , Precursores de Proteínas/análisis , alfa 1-Antitripsina/análisis
17.
Food Chem ; 327: 127002, 2020 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-32438262

RESUMEN

This study was designed to investigate proteome changes in Japanese puffer fish (Takifugu rubripes) during short- and long-term frozen storage. In total, 1484 proteins were quantified, and 164 proteins were identified as differential abundance proteins (DAPs) in Japanese puffer fish from two frozen storage treatment groups (14 days and 60 days) compared with the fresh control group. Correlation analysis between the DAPs and quality traits of the puffer fish muscle showed that 106 proteins were correlated closely with colour and texture (hardness, elasticity, and chewiness). Bioinformatics analysis revealed and Western blot analysis verified that Putative prothymosin alpha species, Bridging integrator 3, NADH: the ubiquinone oxidoreductase subunit and Mx species are candidate biomarkers for puffer fish properties. This study offers valuable evidence to improve the quality control and monitoring of Japanese puffer fish during transportation and storage.


Asunto(s)
Biomarcadores/análisis , Productos Pesqueros/análisis , Proteínas de Peces/análisis , Takifugu , Animales , Color , Biología Computacional/métodos , Análisis de los Alimentos/métodos , Calidad de los Alimentos , Almacenamiento de Alimentos , Congelación , Músculo Esquelético/química , Precursores de Proteínas/análisis , Timosina/análogos & derivados , Timosina/análisis
18.
Proc Natl Acad Sci U S A ; 117(19): 10609-10613, 2020 05 12.
Artículo en Inglés | MEDLINE | ID: mdl-32341146

RESUMEN

Autism spectrum disorder (ASD) is a brain disorder characterized by social impairments. ASD is currently diagnosed on the basis of behavioral criteria because no robust biomarkers have been identified. However, we recently found that cerebrospinal fluid (CSF) concentration of the "social" neuropeptide arginine vasopressin (AVP) is significantly lower in pediatric ASD cases vs. controls. As an initial step in establishing the direction of causation for this association, we capitalized upon a rare biomaterials collection of newborn CSF samples to conduct a quasi-prospective test of whether this association held before the developmental period when ASD first manifests. CSF samples had been collected in the course of medical care of 0- to 3-mo-old febrile infants (n = 913) and subsequently archived at -70 °C. We identified a subset of CSF samples from individuals later diagnosed with ASD, matched them 1:2 with appropriate controls (n = 33 total), and quantified their AVP and oxytocin (OXT) concentrations. Neonatal CSF AVP concentrations were significantly lower among ASD cases than controls and individually predicted case status, with highest precision when cases with comorbid attention-deficit/hyperactivity disorder were removed from the analysis. The associations were specific to AVP, as ASD cases and controls did not differ in neonatal CSF concentrations of the structurally related neuropeptide, OXT. These preliminary findings suggest that a neurochemical marker of ASD may be present very early in life, and if replicated in a larger, prospective study, this approach could transform how ASD is detected, both in behaviorally symptomatic children, and in infants at risk for developing it.


Asunto(s)
Trastorno del Espectro Autista/diagnóstico , Trastorno Autístico/diagnóstico , Vasopresinas/análisis , Arginina Vasopresina/análisis , Arginina Vasopresina/líquido cefalorraquídeo , Trastorno del Espectro Autista/líquido cefalorraquídeo , Trastorno Autístico/líquido cefalorraquídeo , Biomarcadores/líquido cefalorraquídeo , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Registros Médicos , Neuropéptidos , Neurofisinas/análisis , Neurofisinas/líquido cefalorraquídeo , Oxitocina , Estudios Prospectivos , Precursores de Proteínas/análisis , Precursores de Proteínas/líquido cefalorraquídeo , Conducta Social , Vasopresinas/líquido cefalorraquídeo
19.
FEBS Open Bio ; 10(3): 444-454, 2020 03.
Artículo en Inglés | MEDLINE | ID: mdl-31953987

RESUMEN

The insertion of membrane proteins requires proteinaceous complexes in the cytoplasm, the membrane, and the lumen of organelles. Most of the required complexes have been described, while the components for insertion of ß-barrel-type proteins into the outer membrane of chloroplasts remain unknown. The same holds true for the signals required for the insertion of ß-barrel-type proteins. At present, only the processing of Toc75-III, the ß-barrel-type protein of the central chloroplast translocon with an atypical signal, has been explored in detail. However, it has been debated whether Toc75-V/ outer envelope protein 80 (OEP80), a second protein of the same family, contains a signal and undergoes processing. To substantiate the hypothesis that Toc75-V/OEP80 is processed as well, we reinvestigated the processing in a protoplast-based assay as well as in native membranes. Our results confirm the existence of a cleavable segment. By protease protection and pegylation, we observed intermembrane space localization of the soluble N-terminal domain. Thus, Toc75-V contains a cleavable N-terminal signal and exposes its polypeptide transport-associated domains to the intermembrane space of plastids, where it likely interacts with its substrates.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Cloroplastos/metabolismo , Proteínas de la Membrana/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Cloroplastos/química , Cloroplastos/genética , Citoplasma/metabolismo , Membranas Intracelulares/química , Membranas Intracelulares/metabolismo , Proteínas de la Membrana/genética , Membranas/metabolismo , Mitocondrias/metabolismo , Membrana Nuclear/metabolismo , Pisum sativum/genética , Pisum sativum/metabolismo , Precursores de Proteínas/análisis , Precursores de Proteínas/metabolismo , Transporte de Proteínas
20.
Analyst ; 145(4): 1302-1309, 2020 Feb 17.
Artículo en Inglés | MEDLINE | ID: mdl-31913374

RESUMEN

Lamellar MoS2 nanosheets were successfully prepared by hydrothermal synthesis using 1-(3-mercaptopropyl)-3-methyl-imidazolium bromine (MIMBr) ionic liquid as a sulfur source and a morphology control agent, and sodium molybdate as a molybdenum source. Gold nanoparticles were assembled on the surface of MoS2 nanosheets by the in situ reduction of chloroauric acid at low temperatures to successfully fabricate AuNP/2D-MoS2 nanocomposites, thus improving photoelectrochemical response. AuNP/2D-MoS2 nanocomposites were used as photoelectrically active materials modified onto a glassy carbon electrode surface to construct a photoelectrochemical (PEC) sensor. Then, using 1-(N-pyrrolpropyl)-3-methyl-imidazolium bromine (PMIMBr) ionic liquid as a functional monomer and pro-gastrin-releasing peptide (Pro-GRP) as a template, a molecularly imprinted polymerized ionic liquid film was electrochemically deposited on an AuNP/2D-MoS2/GCE surface. Upon removing the templates, a molecularly imprinted photoelectrochemical sensor was constructed for the sensing of a tumor marker, pro-gastrin-releasing peptide. Experimental conditions including ascorbic acid concentration, polymerization conditions, incubation time, and pH value of the incubation solution have been optimized. Under the optimized conditions, the molecularly imprinted photoelectrochemical sensor can specifically detect the target protein Pro-GRP in the range of 0.02 ng mL-1-5 ng mL-1 with a detection limit of 0.0032 ng mL-1 (S/N = 3). The practicability of this photoelectrochemical sensor was demonstrated by accurately determining Pro-GRP in human serum samples.


Asunto(s)
Disulfuros/química , Electroquímica/instrumentación , Oro/química , Nanopartículas del Metal/química , Impresión Molecular , Molibdeno/química , Procesos Fotoquímicos , Precursores de Proteínas/análisis , Células HeLa , Humanos , Concentración de Iones de Hidrógeno , Líquidos Iónicos/química , Límite de Detección , Nanocompuestos/química , Propiedades de Superficie
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