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1.
Proc Natl Acad Sci U S A ; 120(42): e2309007120, 2023 10 17.
Artigo em Inglês | MEDLINE | ID: mdl-37812708

RESUMO

Phytohormone auxin plays a key role in regulating plant organogenesis. However, understanding the complex feedback signaling network that involves at least 29 proteins in Arabidopsis in the dynamic context remains a significant challenge. To address this, we transplanted an auxin-responsive feedback circuit responsible for plant organogenesis into yeast. By generating dynamic microfluidic conditions controlling gene expression, protein degradation, and binding affinity of auxin response factors to DNA, we illuminate feedback signal processing principles in hormone-driven gene expression. In particular, we recorded the regulatory mode shift between stimuli counting and rapid signal integration that is context-dependent. Overall, our study offers mechanistic insights into dynamic auxin response interplay trackable by synthetic gene circuits, thereby offering instructions for engineering plant architecture.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Retroalimentação , Genes Sintéticos , Arabidopsis/metabolismo , Ácidos Indolacéticos/metabolismo , Regulação da Expressão Gênica de Plantas
2.
Ann Rheum Dis ; 82(5): 630-638, 2023 05.
Artigo em Inglês | MEDLINE | ID: mdl-36737106

RESUMO

OBJECTIVE: Neutrophil infiltration into the synovial joint is a hallmark of rheumatoid arthritis (RA), a disease characterised by progressive bone erosion. However, the mechanisms by which neutrophils participate in bone destruction remain unclear. Carbamylation is a posttranslational modification linked to increased bone erosion in RA and we previously showed that carbamylation is present in RA neutrophil extracellular traps (NETs). However, it remains unclear whether NETs and their carbamylated protein cargo directly promote bone destruction and alter osteoclast biology. METHODS: NETs and carbamylated NETs (cNETs) were assessed for their capacity to induce osteoclast formation in CD14+ monocytes. Chemical inhibitors and neutralising antibodies were used to elucidate the pathway by which NETs induce osteoclastogenesis. HLA-DRB1*04:01 mice received intra-articular injection of cNETs for 4 weeks. Joints were isolated and assessed for osteoclast formation. Plasma and synovial fluid samples from patients with RA (n=32) were assessed for the presence of carbamylated histone, and correlations to disease specific outcomes were performed. RESULTS: We found that NETs, when cNETs, instruct monocytes to undergo rapid osteoclast formation. NET-mediated osteoclastogenesis appears to depend on Toll-like receptor 4 signalling and NET-associated proteins including histones and neutrophil elastase. In vivo, we identified that the number of osteoclasts increased following immunisation with cNETs in HLA-DRB1*04:01 transgenic mice. Furthermore, carbamylated histones are increased in plasma and synovial fluid from patients with RA and correlate with active bone resorption and inflammatory markers. CONCLUSIONS: Our results suggest that NETs have a direct role in RA-associated bone erosion by promoting osteoclast formation.


Assuntos
Artrite Reumatoide , Armadilhas Extracelulares , Camundongos , Animais , Histonas , Osteoclastos , Armadilhas Extracelulares/metabolismo , Carbamilação de Proteínas
3.
Elife ; 112022 11 09.
Artigo em Inglês | MEDLINE | ID: mdl-36350114

RESUMO

Cells convert electrical signals into chemical outputs to facilitate the active transport of information across larger distances. This electrical-to-chemical conversion requires a tightly regulated expression of ion channels. Alterations of ion channel expression provide landmarks of numerous pathological diseases, such as cardiac arrhythmia, epilepsy, or cancer. Although the activity of ion channels can be locally regulated by external light or chemical stimulus, it remains challenging to coordinate the expression of ion channels on extended spatial-temporal scales. Here, we engineered yeast Saccharomyces cerevisiae to read and convert chemical concentrations into a dynamic potassium channel expression. A synthetic dual-feedback circuit controls the expression of engineered potassium channels through phytohormones auxin and salicylate to produce a macroscopically coordinated pulses of the plasma membrane potential. Our study provides a compact experimental model to control electrical activity through gene expression in eukaryotic cell populations setting grounds for various cellular engineering, synthetic biology, and potential therapeutic applications.


Assuntos
Canais Iônicos , Canais de Potássio , Canais Iônicos/genética , Canais Iônicos/metabolismo , Canais de Potássio/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Fenômenos Fisiológicos Celulares , Eletrofisiologia
4.
Rev. colomb. gastroenterol ; 37(3): 276-281, jul.-set. 2022. tab
Artigo em Espanhol | LILACS-Express | LILACS | ID: biblio-1408036

RESUMO

Resumen Objetivos: en Colombia se ha venido implementando la sedación por médicos no anestesiólogos para procedimientos endoscópicos fuera del quirófano. Se describió la experiencia en la unidad de gastroenterología de una clínica de alto nivel de atención en Cali, Colombia. Materiales y métodos: estudio observacional, de tipo cohorte analítica para describir la frecuencia y el tipo de eventos adversos asociados a los procedimientos de sedación por médicos generales, y evaluar los factores asociados a su ocurrencia en pacientes que acudieron a la unidad de endoscopia de la Fundación Valle del Lili para la realización de estudios endoscópicos bajo sedación intravenosa que, por ser de bajo riesgo, fue aplicada por un médico no anestesiólogo entre noviembre de 2018 y junio de 2019. Se realizó análisis descriptivo, se calcularon mediana y rango intercuartílico para las variables numéricas, y frecuencias para las variables cualitativas. Resultados: se incluyeron 1506 participantes, 59,4 % ASA I y 40,6 % ASA II. En promedio, la dosis inicial de propofol fue de 60 mg y la dosis total, de 140 mg. Se registraron eventos adversos no serios en 46 pacientes (3,05 %) y el más común fue la desaturación transitoria (80,4 %). Ningún paciente presentó eventos adversos serios. El puntaje inicial promedio de la escala de Aldrete fue 8, mientras que al alta el puntaje promedio fue de 10. Conclusiones: la sedación para procedimientos endoscópicos dada por médicos no anestesiólogos es segura, siempre y cuando sea realizado por personal entrenado que realice una adecuada valoración de los antecedentes (cardiovasculares, gastrointestinales y neurológicos) y factores de riesgo del paciente dentro del marco de los lineamientos institucionales vigentes.


Abstract Objectives: in Colombia, sedation by non-anesthesiologists for endoscopic procedures outside the operating room has been implemented. A description of an experience in the gastroenterology unit of a tertiary referral hospital in Cali, Colombia, was conducted. Materials and methods: an analytical cohort observational study to describe the frequency and type of adverse events associated with sedation procedures performed by general practitioners and evaluate the factors related to their occurrence in patients who attended the endoscopy unit of Fundación Valle del Lili for endoscopic studies under intravenous sedation. Between November 2018 and June 2019, non-anesthesiologist physicians performed this procedure due to the minimal risk implied. A descriptive analysis was completed, and the median and interquartile range were calculated for numerical variables and frequencies for qualitative variables. Results: There were 1506 participants, 59.4% ASA I and 40.6% ASA II in this study. On average, the starting dose of propofol was 60 mg, and the total dose was 140 mg. Forty-six patients (3.05%) reported non-severe adverse events; the most common occurrence was transient desaturation (80.4%). No patients experienced severe adverse events. The average initial Aldrete scale score was 8, while at discharge, the average score was 10. Conclusions: sedation for endoscopic procedures performed by non-anesthesiologists is safe provided that it is performed by trained personnel conducting a correct assessment of the patient's (cardiovascular, gastrointestinal, and neurological) history and risk factors within the framework of the current institutional guidelines.

5.
Nat Commun ; 12(1): 4017, 2021 06 29.
Artigo em Inglês | MEDLINE | ID: mdl-34188048

RESUMO

The synchronization is a recurring phenomenon in neuroscience, ecology, human sciences, and biology. However, controlling synchronization in complex eukaryotic consortia on extended spatial-temporal scales remains a major challenge. Here, to address this issue we construct a minimal synthetic system that directly converts chemical signals into a coherent gene expression synchronized among eukaryotic communities through rate-dependent hysteresis. Guided by chemical rhythms, isolated colonies of yeast Saccharomyces cerevisiae oscillate in near-perfect synchrony despite the absence of intercellular coupling or intrinsic oscillations. Increased speed of chemical rhythms and incorporation of feedback in the system architecture can tune synchronization and precision of the cell responses in a growing cell collectives. This synchronization mechanism remain robust under stress in the two-strain consortia composed of toxin-sensitive and toxin-producing strains. The sensitive cells can maintain the spatial-temporal synchronization for extended periods under the rhythmic toxin dosages produced by killer cells. Our study provides a simple molecular framework for generating global coordination of eukaryotic gene expression through dynamic environment.


Assuntos
Ciclo Celular/fisiologia , Regulação Fúngica da Expressão Gênica/genética , Saccharomyces cerevisiae/metabolismo , Fenômenos Bioquímicos , Expressão Gênica/genética , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , Saccharomyces cerevisiae/genética
6.
Clin Biochem ; 88: 49-55, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33307060

RESUMO

OBJECTIVES: All patients who undergo cardiopulmonary bypass (CPB) experience some degree of ischemia reperfusion injury (IRI). Severe IRI-induced acute kidney injury (AKI) occurs in approximately 1-2% of patients undergoing CPB. Previous studies using activity-based protein profiling of urine identified group XV phospholipase A2, PLA2G15/LPLA2, as potentially associated with patients who develop AKI post CPB. The present study examined urinary PLA2G15/LPLA2 activity during CPB and in the near postoperative period for associations with subsequent AKI development. DESIGN & METHODS: Samples were collected in a nested case controlled cohort of 21 patients per group who either did (AKI) or did not (non-AKI) develop AKI post-operatively. Serum and urine samples from each patient before, during and after CPB were assayed for PLA2G15/LPLA2 activity. RESULTS: Urine activity significantly increased during the intra operative period. In contrast the activities in paired sera were markedly decreased during CPB. There was no correlation between the serum and urine activity levels of patients. There were no significant differences in activity levels of PLA2G15/LPLA2 in the urine or sera from patients that did and did not develop AKI. CONCLUSIONS: The lack of correlation between serum and urine activity levels suggests that the rapid intraoperative increases in PLA2G15/LPLA2 activity may originate from the kidney and as such offer an intraoperative indicator of early renal response to CPB associated stressors.


Assuntos
Injúria Renal Aguda/enzimologia , Aciltransferases/sangue , Aciltransferases/urina , Ponte Cardiopulmonar , Fosfolipases A2/sangue , Fosfolipases A2/urina , Injúria Renal Aguda/etiologia , Injúria Renal Aguda/patologia , Idoso , Biomarcadores/sangue , Biomarcadores/urina , Ponte Cardiopulmonar/efeitos adversos , Estudos de Casos e Controles , Estudos de Coortes , Feminino , Humanos , Período Intraoperatório , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/enzimologia , Complicações Pós-Operatórias/etiologia , Complicações Pós-Operatórias/patologia , Período Pós-Operatório , Fatores de Risco
7.
Clin Proteomics ; 17: 23, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32549867

RESUMO

BACKGROUND: The pathophysiology of subclinical versus clinical rejection remains incompletely understood given their equivalent histological severity but discordant graft function. The goal was to evaluate serine hydrolase enzyme activities to explore if there were any underlying differences in activities during subclinical versus clinical rejection. METHODS: Serine hydrolase activity-based protein profiling (ABPP) was performed on the urines of a case control cohort of patients with biopsy confirmed subclinical or clinical transplant rejection. In-gel analysis and affinity purification with mass spectrometry were used to demonstrate and identify active serine hydrolase activity. An assay for proteinase 3 (PR3/PRTN3) was adapted for the quantitation of activity in urine. RESULTS: In-gel ABPP profiles suggested increased intensity and diversity of serine hydrolase activities in urine from patients undergoing subclinical versus clinical rejection. Serine hydrolases (n = 30) were identified by mass spectrometry in subclinical and clinical rejection patients with 4 non-overlapping candidates between the two groups (i.e. ABHD14B, LTF, PR3/PRTN3 and PRSS12). Western blot and the use of a specific inhibitor confirmed the presence of active PR3/PRTN3 in samples from patients undergoing subclinical rejection. Analysis of samples from normal donors or from several serial post-transplant urines indicated that although PR3/PRTN3 activity may be highly associated with low-grade subclinical inflammation, the enzyme activity was not restricted to this patient group. CONCLUSIONS: There appear to be limited qualitative and quantitative differences in serine hydrolase activity in patients with subclinical versus clinical renal transplant rejection. The majority of enzymes identified were present in samples from both groups implying that in-gel quantitative differences may largely relate to the activity status of shared enzymes. However qualitative compositional differences were also observed indicating differential activities. The PR3/PRTN3 analyses indicate that the activity status of urine in transplant patients is dynamic possibly reflecting changes in the underlying processes in the transplant. These data suggest that differential serine hydrolase pathways may be active in subclinical versus clinical rejection which requires further exploration in larger patient cohorts. Although this study focused on PR3/PRTN3, this does not preclude the possibility that other enzymes may play critical roles in the rejection process.

8.
Transplantation ; 103(9): 1790-1798, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-30985576

RESUMO

Enzyme activity may be more pathophysiologically relevant than enzyme quantity and is regulated by changes in conformational status that are undetectable by traditional proteomic approaches. Further, enzyme activity may provide insights into rapid physiological responses to inflammation/injury that are not dependent on de novo protein transcription. Activity-based protein profiling (ABPP) is a chemical proteomic approach designed to characterize and identify active enzymes within complex biological samples. Activity probes have been developed to interrogate multiple enzyme families with broad applicability, including but not limited to serine hydrolases, cysteine proteases, matrix metalloproteases, nitrilases, caspases, and histone deacetylases. The goal of this overview is to describe the overall rationale, approach, methods, challenges, and potential applications of ABPP to transplantation research. To do so, we present a case example of urine serine hydrolase ABPP in kidney transplant rejection to illustrate the utility and workflow of this analytical approach. Ultimately, developing novel transplant therapeutics is critically dependent on understanding the pathophysiological processes that result in loss of transplant function. ABPP offers a new dimension for characterizing dynamic changes in clinical samples. The capacity to identify and measure relevant enzyme activities provides fresh opportunities for understanding these processes and may help identify markers of disease activity for the development of novel diagnostics and real-time monitoring of patients. Finally, these insights into enzyme activity may also help to identify new transplant therapeutics, such as enzyme-specific inhibitors.


Assuntos
Ensaios Enzimáticos Clínicos , Rejeição de Enxerto/diagnóstico , Transplante de Rim/efeitos adversos , Análise Serial de Proteínas , Proteômica , Serina Endopeptidases/urina , Animais , Biomarcadores/urina , Rejeição de Enxerto/imunologia , Rejeição de Enxerto/urina , Humanos , Valor Preditivo dos Testes , Resultado do Tratamento , Urinálise , Fluxo de Trabalho
9.
J Proteome Res ; 17(10): 3547-3556, 2018 10 05.
Artigo em Inglês | MEDLINE | ID: mdl-30192561

RESUMO

The processes involved in the initiation of acute kidney injury (AKI) following cardiopulmonary bypass (CPB) are thought to occur during the intraoperative period. Such a rapid development might indicate that some of the inductive events are not dependent on de novo protein synthesis, raising the possibility that changes in activities of pre-existing enzymes could contribute to the development of AKI. Activity-based protein profiling (ABPP) was used to compare the serine hydrolase enzyme activities present in the urines of CPB patients who subsequently developed AKI versus those who did not (non-AKI) during the intra- and immediate postoperative periods. Sequential urines collected from a nested case-control cohort of AKI and non-AKI patients were reacted with a serine hydrolase activity probe, fluorophosphonate-TAMRA, and separated by SDS-PAGE. The patterns and levels of probe-labeled proteins in the two groups were initially comparable. However, within 1 h of CPB there were significant pattern changes in the AKI group. Affinity purification and mass spectrometry-based analysis of probe-labeled enzymes in AKI urines at 1 h CPB and arrival to the intensive care unit (ICU) identified 28 enzymes. Quantitative analysis of the activity of one of the identified enzymes, kallikrein-1, revealed some trends suggesting differences in the levels and temporal patterns of enzyme activity between a subset of patients who developed AKI and those who did not. A comparative analysis of affinity-purified probe reacted urinary proteins from these patient groups during the intraoperative period suggested the presence of both shared and unique enzyme patterns. These results indicate that there are intraoperative changes in the levels and types of serine hydrolase activities in patients who subsequently develop AKI. However, the role of these activity differences in the development of AKI remains to be determined.


Assuntos
Injúria Renal Aguda/metabolismo , Ponte Cardiopulmonar/métodos , Hidrolases/metabolismo , Proteômica/métodos , Serina/metabolismo , Injúria Renal Aguda/etiologia , Injúria Renal Aguda/urina , Idoso , Ponte Cardiopulmonar/efeitos adversos , Estudos de Casos e Controles , Feminino , Humanos , Hidrolases/urina , Período Intraoperatório , Masculino , Espectrometria de Massas/métodos , Pessoa de Meia-Idade , Calicreínas Teciduais/metabolismo
10.
Clin Proteomics ; 13: 17, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27528862

RESUMO

BACKGROUND: The urinary proteome of patients undergoing cardiopulmonary bypass (CPB) may provide important insights into systemic and renal changes associated with the procedure. Such information may ultimately provide a basis to differentiate changes or properties associated with the development of acute kidney injury. While mass spectrometry (MS) analysis offers the potential for in-depth compositional analysis it is often limited in coverage and relative quantitation capacity. The aim of this study was to develop a process flow for the preparation and comparison of the intraoperative urinary proteome. METHODS: Urines were collected from patients at the start of CPB and 1-h into CPB. Pooled samples (n = 5) from each time point were processed using a modified Filter Assisted Sample Preparation protocol. The resulting peptides were analyzed by 2D-LC-MS/MS and by 1D-LC-MS/MS SWATH (Sequential Window acquisition of All Theoretical fragment ion spectra). RESULTS: The 2D-LC-MS/MS analysis identified 1324 proteins in the two pools, of which 744 were quantifiable. The SWATH approach provided quantitation for 730 proteins, 552 of which overlapped with the common population from the 2D-IDA results. Intensity correlation filtering between the two methods gave 475 proteins for biological interpretation. Proteins displaying greater than threefold changes (>log2 1.59) at 1-hour CPB relative to the initiation of CPB (26 down-regulated and 22 up-regulated) were selected for further analysis. Up-regulated proteins were enriched in GO terms related to humoral immune response, predominantly innate immunity (C4b, lactotransferrin, protein S100-A8, cathelicidin, myeloperoxidase) and extracellular matrix reorganization (e.g. MMP-9). CONCLUSIONS: This study describes a scheme for processing urine from patients undergoing CPB for mass spectrometry-based analysis. The introduction of SWATH into the workflow offers a sample and instrument sparing approach to obtaining consistent in-depth sample analysis. The design of the methodology is such that it can be readily applied to large numbers of clinical samples with the potential for automation. The results also suggest that activation of the innate immune responses occur during cardiac bypass surgery.

11.
Clin Proteomics ; 10(1): 17, 2013 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-24237849

RESUMO

BACKGROUND: Serine hydrolases constitute a large enzyme family involved in a diversity of proteolytic and metabolic processes which are essential for many aspects of normal physiology. The roles of serine hydrolases in renal function are largely unknown and monitoring their activity may provide important insights into renal physiology. The goal of this study was to profile urinary serine hydrolases with activity-based protein profiling (ABPP) and to perform an in-depth compositional analysis. METHODS: Eighteen healthy individuals provided random, mid-stream urine samples. ABPP was performed by reacting urines (n = 18) with a rhodamine-tagged fluorophosphonate probe and visualizing on SDS-PAGE. Active serine hydrolases were isolated with affinity purification and identified on MS-MS. Enzyme activity was confirmed with substrate specific assays. A complementary 2D LC/MS-MS analysis was performed to evaluate the composition of serine hydrolases in urine. RESULTS: Enzyme activity was closely, but not exclusively, correlated with protein quantity. Affinity purification and MS/MS identified 13 active serine hydrolases. The epithelial sodium channel (ENaC) and calcium channel (TRPV5) regulators, tissue kallikrein and plasmin were identified in active forms, suggesting a potential role in regulating sodium and calcium reabsorption in a healthy human model. Complement C1r subcomponent-like protein, mannan binding lectin serine protease 2 and myeloblastin (proteinase 3) were also identified in active forms. The in-depth compositional analysis identified 62 serine hydrolases in urine independent of activity state. CONCLUSIONS: This study identified luminal regulators of electrolyte homeostasis in an active state in the urine, which suggests tissue kallikrein and plasmin may be functionally relevant in healthy individuals. Additional serine hydrolases were identified in an active form that may contribute to regulating innate immunity of the urinary tract. Finally, the optimized ABPP technique in urine demonstrates its feasibility, reproducibility and potential applicability to profiling urinary enzyme activity in different renal physiological and pathophysiological conditions.

12.
Appl Microbiol Biotechnol ; 93(2): 723-41, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22159660

RESUMO

Sugar beet pulp is a natural carbon source composed mainly of pectin and cellulose, which is utilized and degraded by the ascomycete Penicillium purpurogenum. The fungus also grows on and degrades acetylated xylan which lacks cellulose and pectin. Both carbon sources have been used in our laboratory to grow the fungus and to purify different enzymes secreted to the medium. The enzymes involved in the complex process of degradation of these carbon sources by the fungus have been explored previously under non-denaturing conditions; multienzyme complexes were separated and some subunits identified by Western blots and mass spectrometry. In this work, proteomic profiles show that the secretome is composed of numerous proteins varying in pI and molecular weight. Some enzymes are common to both growth conditions, while others are specific for each carbon source. The results show that the carbon sources utilized exert strong regulatory control over the proteins secreted. This is the first secretome study from a lignocellulolytic Penicillium.


Assuntos
Beta vulgaris/química , Enzimas/metabolismo , Perfilação da Expressão Gênica , Pectinas/metabolismo , Penicillium/enzimologia , Penicillium/genética , Xilanos/metabolismo , Celulose/metabolismo , Enzimas/química , Proteínas Fúngicas/análise , Proteínas Fúngicas/química , Ponto Isoelétrico , Peso Molecular , Proteoma/análise
13.
Appl Microbiol Biotechnol ; 89(1): 145-55, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20972675

RESUMO

Proteins secreted by filamentous fungi play key roles in different aspects of their biology. The fungus Penicillium purpurogenum, used as a model organism, is able to degrade hemicelluloses and pectins by secreting a variety of enzymes to the culture medium. This work shows that these enzymes interact with each other to form high molecular weight, catalytically active complexes. By using a proteomics approach, we were able to identify several protein complexes in the secretome of this fungus. The expression and assembly of these complexes depend on the carbon source used and display molecular masses ranging from 300 to 700 kDa. These complexes are composed of a variety of enzymes, including arabinofuranosidases, acetyl xylan esterases, feruloyl esterases, ß-glucosidases and xylanases. The protein-protein interactions in these multienzyme complexes were confirmed by coimmunoprecipitation assays. One of the complexes was purified from sugar beet pulp cultures and the subunits identified by tandem mass spectrometry. A better understanding of the biological significance of these kinds of interactions will help in the comprehension of the degradation mechanisms used by fungi and may be of special interest to the biotechnology industry.


Assuntos
Proteínas Fúngicas/química , Complexos Multienzimáticos/química , Penicillium/enzimologia , Proteômica , Via Secretória , Sequência de Aminoácidos , Eletroforese em Gel de Poliacrilamida , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Espectrometria de Massas , Dados de Sequência Molecular , Peso Molecular , Complexos Multienzimáticos/genética , Complexos Multienzimáticos/metabolismo , Penicillium/química , Penicillium/genética
14.
Neurosci Lett ; 448(1): 115-9, 2008 Dec 19.
Artigo em Inglês | MEDLINE | ID: mdl-18852024

RESUMO

Prenatal undernutrition induces a variety of cardiovascular alterations in mammals when adults, including hypertension and hypercortisolism, which are thought to be caused by decreased glucocorticoid feedback control of the hypothalamus-pituitary-adrenal (HPA) axis programmed during fetal life. Hypothalamic CRH seems to be involved in blood pressure elevation of spontaneously hypertensive rats and in primary hypertension of humans, but the influence of prenatal undernutrition on CRH expression has deserved little attention. Here, we studied the expression of both CRH mRNA and CRH protein in the hypothalamus of neonatal and juvenile offspring of rats undernourished during fetal life, as well as the plasma levels of CRH and corticosterone. Prenatal undernutrition of pups was induced by submitting pregnant rats to diet restriction (10g daily of 21% protein standard laboratory diet). Pups born from dams with free access to the standard laboratory diet served as controls. At day 2 of postnatal age, undernourished pups showed lower body and brain weights, but higher plasma CRH and corticosterone than normal pups. At day 40 of age, brain weight was significantly decreased in the undernourished rats, while plasma corticosterone, plasma CRH and systolic pressure were significantly increased in these animals. At days 2 and 40 of postnatal age, increased CRH mRNA expression and CRH concentration were found in the hypothalamus of undernourished rats. Results indicate that, in the rat, prenatal undernutrition led to fetal programming of CRH overexpression, a neuropeptide serving as activating signal to the HPA axis and/or to extrahypothalamic brain regions concerned with cardiovascular regulation.


Assuntos
Corticosterona/sangue , Hormônio Liberador da Corticotropina/genética , Hormônio Liberador da Corticotropina/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Hipotálamo/crescimento & desenvolvimento , Hipotálamo/fisiopatologia , Desnutrição/patologia , Fatores Etários , Animais , Animais Recém-Nascidos , Peso Corporal/fisiologia , Modelos Animais de Doenças , Feminino , Masculino , Desnutrição/metabolismo , Desnutrição/fisiopatologia , Gravidez , Ratos , Ratos Wistar
15.
Neurosci Lett ; 419(2): 99-103, 2007 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-17481809

RESUMO

Prenatal undernutrition is known to disturb the hypothalamo-pituitary-adrenal (HPA) axis, possibly through the programming of decreased expression of hypothalamic and pituitary glucocorticoid receptors. To test this hypothesis, we examined the corticosterone response to moderate subcutaneous (100 microg/kg) and intra-paraventricular (50 pmol, bilaterally) dexamethasone (DEX) challenges in normal eutrophic and prenatally undernourished young rats. Undernutrition was induced during fetal life by restricting the diet of pregnant mothers to 10 g daily, while mothers of eutrophic rats received the same diet ad libitum. At day 40 of postnatal life (i) undernourished rats showed increased plasma corticosterone concentration compared to normals; and (ii) subcutaneous and intra-paraventricular administrations of DEX led to reduced corticosterone levels in normal and undernourished animals, the effect of DEX (administered either peripherally or centrally) being significantly lower in the latter group. Results suggest that the low sensitivity of the HPA axis to DEX as well as the increased plasma corticosterone observed in prenatally undernourished rats could be due to the already reported glucocorticoid receptor underexpression found in the hypothalamus and pituitary of in utero undernourished animals, but alternative explanations involving central noradrenergic adaptive changes could also be possible.


Assuntos
Corticosterona/metabolismo , Doenças do Sistema Endócrino/fisiopatologia , Transtornos da Nutrição Fetal/fisiopatologia , Sistema Hipotálamo-Hipofisário/fisiopatologia , Sistema Hipófise-Suprarrenal/fisiopatologia , Receptores de Glucocorticoides/metabolismo , Envelhecimento/fisiologia , Animais , Animais Recém-Nascidos , Corticosterona/sangue , Dexametasona/farmacologia , Doenças do Sistema Endócrino/etiologia , Feminino , Glucocorticoides/farmacologia , Sistema Hipotálamo-Hipofisário/efeitos dos fármacos , Sistema Hipotálamo-Hipofisário/metabolismo , Injeções Subcutâneas , Masculino , Núcleo Hipotalâmico Paraventricular/efeitos dos fármacos , Núcleo Hipotalâmico Paraventricular/metabolismo , Núcleo Hipotalâmico Paraventricular/fisiopatologia , Sistema Hipófise-Suprarrenal/efeitos dos fármacos , Sistema Hipófise-Suprarrenal/metabolismo , Gravidez , Ratos , Ratos Wistar , Receptores de Glucocorticoides/efeitos dos fármacos
16.
Rev. mex. pediatr ; 64(6): 254-6, nov.-dic. 1997. tab
Artigo em Espanhol | LILACS | ID: lil-225186

RESUMO

El objetivo de este trabajo fue conocer la frecuencia de aislamiento de Salmonella spp. en una cohorte de niños asintomáticos, que asistían a dos guarderias de la ciudad de Mérida: Diecisiete niños fueron seleccionados para el estudio porque previamente se había encontrado en ellos esta bacteria. Se recolectaron muestras fecales para coprocultivo cada 15 días durante seis meses. De las 185 muestras colectadas 33 (17.8 por ciento) fueron positivas a Salmonella; se distribuyeron entre 13 (776.5 por ciento) niños, de éstos, 2(11.8 por ciento) fueron positivos en una sola muestra y 11 (64.7 por ciento) en más de una. Se comenta la posibilidad de que estos niños asintomáticos estuviesen actuando como portadores en la guardería y en el ambiente familiar, sobre todo aquellos que fueron positivos en forma intermitente a lo largo del estudio


Assuntos
Humanos , Lactente , Pré-Escolar , Salmonella/isolamento & purificação , Epidemiologia/estatística & dados numéricos
17.
Vet. Méx ; 27(4): 319-24, oct.-dic. 1996. tab
Artigo em Espanhol | LILACS | ID: lil-208075

RESUMO

Entre los diferentes microorganismos indicadores de contaminación o de prácticas indeseables en el manejo de los alimentos se encuentran las bacterias mesofilicas aerobias, los organismos coliformes y los enterococos. El Vibrio cholerae toxigénico, agente etiológico del cólera, ha sido asociado con numerosos productos de pesca, en especial con los moluscos bivalvos sobre todo si se consumen crudos. Por otro lado, la causa más frecuente de salmonelosis es el consumo de alimentos contaminados. Otros microorganismos patógenos involucrados en los síndromes de intoxicación alimentaria son las cepas de Staphylococcus aureus productoras de enterotoxinas. El objetivo de este trabajo fue conocer la prevalencia de microorganismos mesofílicos aerobios, coliformes, enterococos, Vibrio cholerae, Salmonella y Staphylococcus aureus en los alimentos marinos que se expenden en restaurantes de la ciudad de Mérida, Yucatán. Se estudiaron 284 tipos de alimentos que se seleccionaron de 53 restantes. Para analizarlos se utilizaron técnicas convencionales, y se consideraron las normas sanitarias existentes. Para los alimentos sin normas sanitarias se obtuvo la media del número de colonias aisladas. No se aisló V. cholerae O1 y no O1, pero se aisló V. parahaemolyticus en 6/284 (2.1 por ciento) y V. alginolyticus en 12/284 (4.2 por ciento) de las muestras. Se concluye que los alimentos marinos que se expenden en la ciudad de México no presentan una fuente de infección por V. cholerae, pero el nivel de contaminación tanto ambiental como fecal proveniente posiblemente de los manejadores de alimentos, representa un riesgo importante de adquirir otro microorganismo enteropatógeno


Assuntos
Salmonella/patogenicidade , Frutos do Mar/microbiologia , Staphylococcus aureus/patogenicidade , Vibrio cholerae/patogenicidade , Vibrio parahaemolyticus/isolamento & purificação , Vigilância Sanitária , Produtos Pesqueiros/parasitologia , Cólera/prevenção & controle , Contaminação de Alimentos/prevenção & controle , Poluição da Água/efeitos adversos , Doenças Transmitidas por Alimentos/prevenção & controle
18.
Asunción; Paraguay. Ministerio de Salud Pública; jul. 1996. 64 p. tab, graf.
Monografia em Espanhol, Inglês | LILACS, BDNPAR | ID: lil-264132

RESUMO

Presenta un estudio sobre la percepción y el consumo de servicios de salud y tiene entre sus objetivos la provisión de información que ayude a priorizar las inversiones del programa. Consisten en estudios de infraestructura, de epidemiología y de recursos humanos, todos esfuerzos analíticos desarrollados dentro del marco del programa


Assuntos
Serviços de Saúde , Medição de Consumo de Água , Paraguai
19.
Salud pública Méx ; 37(3): 236-239, mayo-jun. 1995. tab
Artigo em Espanhol | LILACS | ID: lil-167364

RESUMO

Para determinar la calidad microbiológica del agua potable de la ciudad de Mérida, Yucatán, México, se analizaron 383 pares de muestras (dos por domicilio). De éstas, 364 (95 por ciento) muestras de las llaves exteriores y 283 (73.89 por ciento) de las interiores cumplieron las normas microbiológicas. La calidad del agua distribuida es aceptable, excepto en la zona de influencia de la planta Mérida III, en donde se encontró contaminación con mesofílicos aerobios en 21.7 por ciento de las muestras. El agua intradomiciliaria mantiene la calidad y solamente en la zona de influencia de la planta Mérida I se observó contaminación de probable origen fecal en 4.8 por ciento de las muestras


With the aim of knowing the microbiological quality of drinking water in Merida, Yucatan, 383 paired samples of drinking water (two per house) were studied. Three hundred sixty four (95%) city water system samples and 283 (73.89%) tap water samples met the microbiological standards for drinking water. It was concluded that microbiological quality of drinking water from the city water system is satisfactory, except for the water system district Merida III, which has a significant aerobic plate count contamination level (21.7% of the samples). Domestic storage systems preserve water quality, with the exception of district Merida I, which has the highest level of contamination (4.8% of the samples) possibly from sewage water and fecal sources.


Assuntos
Abastecimento de Água , México , Poluição da Água , Microbiologia da Água
20.
Vet. Méx ; 26(1): 37-40, ene.-mar. 1995.
Artigo em Espanhol | LILACS | ID: lil-173869

RESUMO

Se realizó un estudio para conocer la frecuencia de enterotoxinas termolábil (LTh) y termoestable (STa) en cepas de Escherichia coli aisladas de heces de 300 cerdos de 1 a 10 días de edad que cursaron con un cuadro diarreico. Se aisló E. coli en el 100 por ciento de los animales muestreados. Mediante la técnica de ELISA, se detectó la producción de enterotoxina termoestable (STa) en 35 animales (11.6 por ciento). No se encontraron cepas de E. coli productoras de enterotoxina termolábil (LTh). Se recomienda la utilización de antisuero homólogo al realizar la técnica de GM1-ELISA debido a que existen diferencias inmunológicas entre la LT humana (LTh) y la LT porcina (LTp)


Assuntos
Animais , Suínos/microbiologia , Ensaio de Imunoadsorção Enzimática/veterinária , Diarreia/veterinária , Enterotoxinas/fisiologia , Escherichia coli/isolamento & purificação , Soros Imunes
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