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Asian Journal of Andrology ; (6): 238-242, 2022.
Article in English | WPRIM | ID: wpr-928542


Cilium, an organelle with a unique proteome and organization, protruding from the cell surface, generally serves as a force generator and signaling compartment. During ciliogenesis, ciliary proteins are synthesized in cytoplasm and transported into cilia by intraflagellar transport (IFT) particles, where the inner counterparts undergo reverse trafficking. The homeostasis of IFT plays a key role in cilial structure assembly and signaling transduction. Much progress has been made on the mechanisms and functions of IFT; however, recent studies have revealed the involvement of IFT particle subunits in organogenesis and spermatogenesis. In this review, we discuss new concepts concerning the molecular functions of IFT protein IFT25 and how its interactions with other IFT particle subunits are involved in mammalian development and fertility.

Animals , Male , Biological Transport , Carrier Proteins/metabolism , Cilia/metabolism , Flagella/metabolism , Mammals/metabolism , Organogenesis , Proteins/metabolism , Signal Transduction
Chinese Journal of Biotechnology ; (12): 1462-1474, 2022.
Article in Chinese | WPRIM | ID: wpr-927793


Extracellular vesicles (EVs), also known as membrane vesicles, are vesicular bodies secreted by eukaryotic cells and bacteria. EVs can carry proteins, DNA, RNA, and various metabolites for the exchange and transmission of substances between cells. They play contents-dependent physiological functions, such as delivering nutrients, participating in immune response, and treating cancers. Currently, most studies focus on the exploration of vesicles secreted by eukaryotic cells and gram-negative bacteria, while few studies focus on gram-positive bacteria. This review summarized the production, content composition, physiological function, and engineering of EVs secreted by gram-positive bacteria, and prospected future perspectives in this area.

Bacteria/metabolism , Extracellular Vesicles/metabolism , Gram-Negative Bacteria , Gram-Positive Bacteria/metabolism , Proteins/metabolism
Chinese Journal of Biotechnology ; (12): 3915-3932, 2021.
Article in Chinese | WPRIM | ID: wpr-921476


Targeted protein degradation (TPD) technology facilitates specific and efficient degradation of disease-related proteins through hijacking the two major protein degradation systems in mammalian cells: ubiquitin-proteasome system and lysosome pathway. Compared with traditional small molecule-inhibitors, TPD-based drugs exhibit the characteristics of a broader target spectrum. Compared with techniques interfere with protein expression on the gene and mRNA level, TPD-based drugs are target-specific, efficaciously rapid, and not constrained by post-translational modification of proteins. In the past 20 years, various TPD-based technologies have been developed. Most excitingly, two TPD-based therapeutic drugs have been approved by FDA for phase Ⅰ clinical trials in 2019. Despite of the early stage characteristics and various obstructions of the TPD technology, it could serve as a powerful tool for the development of novel drugs. This review summarizes the advances of different degradation systems based on TPD technologies and their applications in disease therapy. Moreover, the advantages and challenges of various technologies were discussed systematically, with the aim to provide theoretical guidance for further application of TPD technologies in scientific research and drug development.

Animals , Proteasome Endopeptidase Complex/metabolism , Protein Processing, Post-Translational , Proteins/metabolism , Proteolysis , Technology
Electron. j. biotechnol ; 37: 18-24, Jan. 2019. tab, ilus, graf
Article in English | LILACS | ID: biblio-1049076


BACKGROUND: The 11S globulin from amaranth is the most abundant storage protein in mature seeds and is well recognized for its nutritional value. We used this globulin to engineer a new protein by adding a four valinetyrosine antihypertensive peptide at its C-terminal end to improve its functionality. The new protein was named AMR5 and expressed in the Escherichia coli BL21-CodonPlus(DE3)-RIL strain using a custom medium (F8PW) designed for this work. RESULTS: The alternative medium allowed for the production of 652 mg/L expressed protein at the flask level, mostly in an insoluble form, and this protein was subjected to in vitro refolding. The spectrometric analysis suggests that the protein adopts a ß/α structure with a small increment of α-helix conformation relative to the native amaranth 11S globulin. Thermal and urea denaturation experiments determined apparent Tm and C1/2 values of 50.4°C and 3.04 M, respectively, thus indicating that the antihypertensive peptide insertion destabilized the modified protein relative to the native one. AMR5 hydrolyzed by trypsin and chymotrypsin showed 14- and 1.3-fold stronger inhibitory activity against angiotensin I-converting enzyme (IC50 of 0.034 mg/mL) than the unmodified protein and the previously reported amaranth acidic subunit modified with antihypertensive peptides, respectively. CONCLUSION: The inserted peptide decreases the structural stability of amaranth 11S globulin and improves its antihypertensive activity.

Peptides/metabolism , Proteins/metabolism , Globulins/metabolism , Antihypertensive Agents/metabolism , Seeds , Temperature , Culture Media , Amaranthus , Protein Stability , Phytochemicals
Rev. méd. Chile ; 147(10): 1323-1328, oct. 2019.
Article in Spanish | LILACS | ID: biblio-1058600


Renal functional reserve (RFR) is the capacity of the kidney to increase its glomerular filtration rate (GFR) in response to physiological or pathological stimuli. The most commonly used stimuli to assess this reserve are an oral load of proteins of animal origin, amino acid infusions, dopamine, glucagon or combinations of them. RFR is calculated as the difference between stimulated and baseline GFR. Vegetarians have lower baseline GFR than the general population and an increased RFR. Subjects with only one kidney and those suffering from chronic nephropathies usually have a reduced or absent RFR despite having normal basal GFR. Quantification of RFR may be useful to detect subclinical renal damage, physiological conditions that reduce baseline GFR, evaluation of potential donors for kidney transplantation, suspected hyperfiltration, detection of renal lability against acute injuries or pregnancy and the evaluation after an acute renal injury when renal function seems to be recovered and residual subclinical damage is suspected.

Humans , Male , Female , Middle Aged , Young Adult , Acute Kidney Injury/physiopathology , Glomerular Filtration Rate/physiology , Proteins/metabolism , Risk Factors , Creatinine/blood , Acute Kidney Injury/metabolism
Arch. argent. pediatr ; 116(4): 515-521, ago. 2018. graf, tab
Article in English, Spanish | LILACS, BINACIS | ID: biblio-950044


Introducción: "Gran quemado" es quien sufre lesiones por daño térmico que afectan más del 30% de su superficie corporal (SC). El hipercatabolismo secundario causa pérdida de masa magra y retraso de la cicatrización de heridas. Objetivo: Describir y analizar los resultados de la implementación de un protocolo de soporte nutricional en niños quemados graves internados en una Unidad de Cuidados Intensivos durante las primeras 6 semanas evolutivas. Población y métodos: Diseño analítico, prospectivo, observacional y longitudinal. Se midieron peso, talla, porcentaje de SC quemada, días de internación en la Unidad de Cuidados Intensivos y mortalidad. Se analizaron tasa metabólica basal por calorimetría indirecta y fórmula de Schofield, cobertura de aporte energético y proteico, prealbúmina, proteína C reactiva, vitaminas A, D, E, cobre y zinc semanales. Resultados: Se incluyeron 18 pacientes (media: 3,9 años, 49% de SC quemada). Se alcanzó la media de objetivo energético en la segunda semana y el requerimiento proteico en la semana 6. Doce pacientes requirieron nutrición parenteral complementaria sin complicaciones. Se hallaron parámetros de hipermetabolismo, que se normalizaron a las 4-6 semanas del ingreso, excepto la proteína C reactiva. Las vitaminas A y E y elementos traza (zinc y cobre) estaban descendidos al ingreso con mejoría posterior. La vitamina D persistió en valores bajos. Un paciente falleció. Conclusiones: La implementación del protocolo permitió lograr el aporte de la totalidad del requerimiento energético; la cobertura del requerimiento proteico se postergó hasta la semana 6. Es necesario hacer hincapié en resolver las limitaciones para alcanzar este último.

Introduction. "Major burn" is used to describe a person who suffers thermal damage affecting more than 30% of his/her total body surface area (TBSA). The secondary hypercatabolism causes lean body mass loss and delayed wound healing. Objective. To describe and analyze the results of implementing a nutritional support protocol for pediatric burn patients hospitalized in the intensive care unit in the first 6 weeks. Population an d methods. Analytical, prospective, observational, and longitudinal design. Weight, height, %TBSA, length of stay in the intensive care unit, and mortality were measured. The basal metabolic rate was measured by indirect calorimetry and the Schofield equation, and protein and energy intake, prealbumin, C-reactive protein, vitamins A, D, E, copper, and zinc levels were analyzed every week. Results. Eighteen patients were included (mean: 3.9 years old, 49%TBSA). The mean energy target was achieved by week 2 and protein requirements were met by week 6. Twelve patients required complementary parenteral nutrition and there were no complications. Hypermetabolism parameters were observed, which returned to normal 4-6 weeks after hospitalization, except for C-reactive protein. Vitamins A and E and trace elements (zinc and copper) were reduced at the time of admission and showed a subsequent improvement. Vitamin D remained low. One patient died. Conclusions. Implementing the protocol was useful to cover the total energy requirement; the coverage of protein requirements was delayed until week 6. It is necessary to focus on solving limitations to achieve the latter.

Humans , Infant, Newborn , Infant , Child, Preschool , Child , Adolescent , Burns/complications , Parenteral Nutrition/methods , Nutritional Support/methods , Nutritional Requirements , Burns/mortality , Energy Intake , Dietary Proteins/administration & dosage , Intensive Care Units, Pediatric , Proteins/metabolism , Prospective Studies , Longitudinal Studies , Hospitalization , Length of Stay
Braz. j. med. biol. res ; 51(3): e7033, 2018. tab, graf
Article in English | LILACS | ID: biblio-889046


In the present study, we successfully demonstrated for the first time the existence of cardiac proteomic differences between non-selectively bred rats with distinct intrinsic exercise capacities. A proteomic approach based on two-dimensional gel electrophoresis coupled to mass spectrometry was used to study the left ventricle (LV) tissue proteome of rats with distinct intrinsic exercise capacity. Low running performance (LRP) and high running performance (HRP) rats were categorized by a treadmill exercise test, according to distance run to exhaustion. The running capacity of HRPs was 3.5-fold greater than LRPs. Protein profiling revealed 29 differences between HRP and LRP rats (15 proteins were identified). We detected alterations in components involved in metabolism, antioxidant and stress response, microfibrillar and cytoskeletal proteins. Contractile proteins were upregulated in the LVs of HRP rats (α-myosin heavy chain-6, myosin light chain-1 and creatine kinase), whereas the LVs of LRP rats exhibited upregulation in proteins associated with stress response (aldehyde dehydrogenase 2, α-crystallin B chain and HSPβ-2). In addition, the cytoskeletal proteins desmin and α-actin were upregulated in LRPs. Taken together, our results suggest that the increased contractile protein levels in HRP rats partly accounted for their improved exercise capacity, and that proteins considered risk factors to the development of cardiovascular disease were expressed in higher amounts in LRP animals.

Animals , Male , Rats , Physical Conditioning, Animal/physiology , Running/physiology , Proteins/metabolism , Heart Function Tests/methods , Myocardium/metabolism , Organ Size , Rats, Inbred Strains , Mass Spectrometry , Electrophoresis, Gel, Two-Dimensional , Proteins/isolation & purification , Contractile Proteins/metabolism , Cytoskeletal Proteins/metabolism , Proteomics , Desmin/metabolism , Heart Ventricles/metabolism , Heat-Shock Proteins/metabolism
Bol. méd. Hosp. Infant. Méx ; 74(3): 175-180, May.-Jun. 2017. graf
Article in English | LILACS | ID: biblio-888613


Abstract: Background: Mitochondriopathies are multisystem diseases affecting the oxidative phosphorylation (OXPHOS) system. Skin fibroblasts are a good model for the study of these diseases. Fibroblasts with a complex IV mitochondriopathy were used to determine the molecular mechanism and the main affected functions in this disease. Methods: Skin fibroblast were grown to assure disease phenotype. Mitochondria were isolated from these cells and their proteome extracted for protein identification. Identified proteins were validated with the MitoMiner database. Results: Disease phenotype was corroborated on skin fibroblasts, which presented a complex IV defect. The mitochondrial proteome of these cells showed that the most affected proteins belonged to the OXPHOS system, mainly to the complexes that form supercomplexes or respirosomes (I, III, IV, and V). Defects in complex IV seemed to be due to assembly issues, which might prevent supercomplexes formation and efficient substrate channeling. It was also found that this mitochondriopathy affects other processes that are related to DNA genetic information flow (replication, transcription, and translation) as well as beta oxidation and tricarboxylic acid cycle. Conclusions: These data, as a whole, could be used for the better stratification of these diseases, as well as to optimize management and treatment options.

Resumen: Introducción: Las mitocondriopatías son enfermedades multisistémicas que afectan el funcionamiento de la fosforilación oxidativa (OXPHOS). Un buen modelo de estudio para estas enfermedades es el cultivo primario de fibroblastos. En este trabajo se utilizaron fibroblastos con mitocondriopatía del complejo IV para determinar cuáles son las principales funciones afectadas en esta enfermedad. Métodos: Se realizaron cultivos primarios de fibroblastos para corroborar el fenotipo de la enfermedad. Las mitocondrias se aislaron de estas células y se extrajo su proteoma para su identificación. Las proteínas identificadas se validaron con la base de datos de MitoMiner. Resultados: Los fibroblastos conservaron el fenotipo de la enfermedad que incluye un defecto del complejo IV. El proteoma mitocondrial de estas células mostró que las proteínas más afectadas pertenecen al sistema de OXPHOS, principalmente los complejos que forman supercomplejos o respirosomas (I, III, IV y V). El defecto en el complejo IV al parecer se debió a problemas de ensamblaje que pueden evitar la formación de los supercomplejos y la eficiente canalización de sustratos. También se observó que esta mitocondriopatía afecta otros procesos relacionados con el flujo de información genética del DNA (replicación, transcripción y traducción), así como con la beta oxidación y el ciclo de los ácidos tricarboxílicos (TCA). Conclusiones: En conjunto, estos datos podrían utilizarse para una mejor clasificación de estas enfermedades, así como para la optimización de las opciones de manejo y tratamiento.

Humans , Cytochrome-c Oxidase Deficiency/pathology , Proteomics/methods , Fibroblasts/pathology , Mitochondria/pathology , Oxidative Phosphorylation , DNA/genetics , Proteins/metabolism , Cells, Cultured , Citric Acid Cycle/physiology
Bol. méd. Hosp. Infant. Méx ; 74(3): 181-192, May.-Jun. 2017. tab, graf
Article in English | LILACS | ID: biblio-888614


Abstract: Introduction: Relapse occurs in approximately 20% of Mexican patients with childhood acute lymphoblastic leukemia (ALL). In this group, chemoresistance may be one of the biggest challenges. An overview of complex cellular processes like drug tolerance can be achieved with proteomic studies. Methods: The B-lineage pediatric ALL cell line CCRF-SB was gradually exposed to the chemotherapeutic vincristine until proliferation was observed at 6 nM, control cells were cultured in the absence of vincristine. The proteome from each group was analyzed by nanoHPLC coupled to an ESI-ion trap mass spectrometer. The identified proteins were grouped into over-represented functional categories with the PANTHER classification system. Results: We found 135 proteins exclusively expressed in the presence of vincristine. The most represented functional categories were: Toll receptor signaling pathway, Ras Pathway, B and T cell activation, CCKR signaling map, cytokine-mediated signaling pathway, and oxidative phosphorylation. Conclusions: Our study indicates that signal transduction and mitochondrial ATP production are essential during adaptation of leukemic cells to vincristine, these processes represent potential therapeutic targets.

Resumen: Introducción: Aproximadamente el 20% de los pacientes mexicanos con leucemia linfoblástica aguda (LLA) infantil presentan recaídas. En este grupo, la quimiorresistencia es uno de los principales desafíos. Los estudios proteómicos pueden dar un panorama general de procesos celulares complejos como la tolerancia a fármacos. Métodos: La línea celular de LLA de linaje B, CCRF-SB, fue expuesta de manera gradual al fármaco quimioterapéutico vincristina hasta observar proliferación celular en presencia de 6 nM, como control se cultivaron células en ausencia del fármaco. Se analizó el proteoma de cada grupo mediante nanoHPLC acoplado a un espectrómetro de masas de tipo trampa de iones. Las proteínas identificadas se agruparon en categorías funcionales sobre-representadas con el sistema de clasificación PANTHER. Resultados: Encontramos 135 proteínas expresadas exclusivamente en presencia de vincristina. Las categorías funcionales más representadas fueron la señalización asociada a los receptores tipo Toll, señalización dependiente de Ras, activación de células B y T, mapa de señalización CCKR, señalización mediada por citoquinas y la fosforilación oxidativa. Conclusiones: Nuestro estudio indica que la transducción de señales y la producción de ATP mitocondrial son procesos esenciales durante la adaptación de células leucémicas a vincristina por lo que estos procesos representan potenciales blancos terapéuticos.

Child , Humans , Vincristine/pharmacology , Proteomics/methods , Precursor Cell Lymphoblastic Leukemia-Lymphoma/drug therapy , Antineoplastic Agents, Phytogenic/pharmacology , Signal Transduction/drug effects , Proteins/metabolism , Gene Expression Regulation, Leukemic , Adenosine Triphosphate/metabolism , Chromatography, High Pressure Liquid , Drug Resistance, Neoplasm , Proteome/metabolism , Spectrometry, Mass, Electrospray Ionization , Cell Line, Tumor , Cell Proliferation/drug effects , Precursor Cell Lymphoblastic Leukemia-Lymphoma/genetics , Mitochondria/metabolism
Rev. cuba. med. mil ; 46(2): 148-162, abr.-jun. 2017. tab
Article in Spanish | LILACS, CUMED | ID: biblio-901213


Introducción: el catabolismo proteico es un indicador de la respuesta metabólica a la agresión. Objetivo: determinar la evolución de los pacientes con ventilación mecánica invasiva y su posible asociación con el catabolismo proteico, por categorías diagnósticas. Método: se realizó un estudio observacional, analítico y prospectivo, conto dos los pacientes sometidos a ventilación mecánica invasiva, ingresados en cuidados intensivos desde el 2001 hasta el 2007 y fueron clasificaron según la categoría diagnóstica (trauma, clínico y quirúrgico). Se midió el peso corporal al ingreso. Se evaluó el catabolismo proteico durante los primeros 3 días del ingreso, con la urea plasmática, la creatininuria y el nitrógeno ureico urinario. Se contrastaron con las variables dependientes: mortalidad, morbilidad y el tiempo de ventilación mecánica. Resultados: se estudiaron 262 pacientes; 88 presentaban trauma, 89 afecciones clínicas y 85 afecciones quirúrgicas. El catabolismo proteico fue alto en el trauma y se asoció a la mortalidad, a la disfunción múltiple de órganos y al tiempo prolongado de ventilación mecánica; en los pacientes quirúrgicos se asoció a la morbilidad. Los valores bajos de creatininuria, evidenciaron asociación con la mayor mortalidad, morbilidad y el tiempo prolongado de ventilación mecánica. Conclusiones: el catabolismo proteico se asoció a la evolución del paciente con ventilación mecánica invasiva, en las categorías trauma y quirúrgicos. No se evidenció asociación en la categoría clínica(AU)

Introduction: Protein catabolism is an indicator of the metabolic response to injury. Objective: To determine the evolution of patients with mechanical invasive ventilation and its possible association with protein catabolism by diagnostic category. Method: An observational, analytical and prospective study was performed with all patients undergoing mechanical invasive ventilation admitted to Intensive Care from 2001 to 2007 and classified according to the diagnostic category (trauma, clinical and surgical). Body weight was measured at admission. Protein catabolism was evaluated during the first 3 days of admission, with plasma urea, creatinine and urinary urea nitrogen. They were contrasted with the dependent variables: mortality, morbidity and time of mechanical ventilation. Results: We studied 262 patients; 88 presented trauma, 89 clinical conditions and 85 surgical conditions. Protein catabolism was high in trauma and associated with mortality, multiple organ dysfunction and prolonged mechanical ventilation; in surgical patients was associated with morbidity. The low values of urine creatinine,were associated with the greater mortality, morbidity and the prolonged time of mechanical ventilation. Conclusions: Protein catabolism was associated with the evolution of the patient with mechanical invasive ventilation, in the trauma and surgical categories. There was no evidence of association in the clinical category(AU)

Humans , Respiration, Artificial/methods , Proteins/metabolism , Critical Care/methods , Respiration, Artificial/mortality , Prospective Studies , Analytical Epidemiology , Observational Study
Electron. j. biotechnol ; 27: 25-31, May. 2017. tab, graf
Article in English | LILACS | ID: biblio-1010164


Background: Rhodotorula glutinis is capable of synthesizing numerous valuable metabolites with extensive potential industrial usage. This paper reports the effect of initial culture medium pH on growth and protein, lipid, and carotenoid biosynthesis by R. glutinis. Results: The highest biomass yield was obtained in media with pH 4.0­7.0, and the value after 72 h was 17.2­19.4 gd.w./L. An initial pH of the medium in the range of 4.0­7.0 has no significant effect on the protein (38.5­41.3 g/100 gd.w.), lipid (10.2­12.7 g/100 gd.w.), or carotenoid (191.7­202.9 µg/gd.w.) content in the biomass or on the profile of synthesized fatty acids and carotenoids. The whole pool of fatty acids was dominated by oleic (48.1­53.4%), linoleic (21.4­25.1%), and palmitic acids (13.0­15.8%). In these conditions, the yeast mainly synthesized torulene (43.5­47.7%) and ß-carotene (34.7­38.6%), whereas the contribution of torularhodin was only 12.1­16.8%. Cultivation in medium with initial pH 3.0 resulted in a reduction in growth (13.0 gd.w./L) and total carotenoid (115.8 µg/gd.w.), linoleic acid (11.5%), and torularhodin (4.5%) biosynthesis. Conclusion: The different values of initial pH of the culture medium with glycerol and deproteinized potato wastewater had a significant effect on the growth and protein, lipid, and carotenoid biosynthesis by R. glutinis.

Rhodotorula/metabolism , Carotenoids/biosynthesis , Yeasts , Solanum tuberosum , Proteins/metabolism , Biomass , Wastewater , Glycerol , Hydrogen-Ion Concentration , Lipids/biosynthesis
Electron. j. biotechnol ; 26: 1-6, Mar. 2017. ilus, graf
Article in English | LILACS | ID: biblio-1008840


Background: Ornithine decarboxylase antizyme 1 (OAZ1) is an important regulator of polyamine synthesis and uptake. Our previous studies indicated that high OAZ1 expression in the ovaries of laying geese is responsible for poor egg production. In the present study, the molecular characterization of goose OAZ1 gene was analyzed, as well as the expression profile in various follicular tissues. Results: An 873-bp cDNA sequence of the OAZ1 gene (Accession No. KC845302) with a +1 frameshift site (+175T) was obtained. The sequence consisted of a 652-bp two overlapping open reading frames (a putative protein with 216 amino acids). The OAZ domain, OAZ signature and OAZ super family domain were prominent conserved regions among species. As the follicle size increased, OAZ1 abundance showed an increasing trend during follicular development, while it decreased during follicular regression. The level of OAZ1 mRNA expression was the lowest in the fifth largest preovulatory follicle, and was 0.65-fold compared to the small white follicle (P b 0.05). OAZ1 mRNA expression in the largest preovulatory and postovulatory follicle was 2.11- and 2.49-fold compared to the small white follicle, respectively (P b 0.05). Conclusions: The goose OAZ1 structure confirms that OAZ1 plays an important role in ornithine decarboxylase-mediated regulation of polyamine homeostasis. Our findings provide an evidence for a potential function of OAZ1 in follicular development, ovulation and regression.

Animals , Female , Proteins/genetics , Proteins/metabolism , Geese/metabolism , Ovarian Follicle/metabolism , Ornithine Decarboxylase/metabolism , Polyamines/metabolism , RNA, Messenger , Cloning, Molecular , Sequence Analysis , DNA, Complementary , Real-Time Polymerase Chain Reaction , Ovarian Follicle/growth & development
Braz. j. med. biol. res ; 50(6): e5661, 2017. tab, graf
Article in English | LILACS | ID: biblio-839315


Burkitt lymphoma (BL) is a highly malignant non-Hodgkin's lymphoma that is closely related to the abnormal expression of genes. Familial acute myelogenous leukemia related factor (FAMLF; GenBank accession No. EF413001.1) is a novel gene that was cloned by our research group, and miR-181b is located in the intron of the FAMLF gene. To verify the role of miR-181b and FAMLF in BL, RNAhybrid software was used to predict target site of miR-181b on FAMLF and real-time quantitative PCR (RQ-PCR) was used to detect expression of miR-181b and FAMLF in BL patients, Raji cells and unaffected individuals. miR-181b was then transfected into Raji and CA46 cell lines and FAMLF expression was examined by RQ-PCR and western blotting. Further, Raji cells viability and proliferation were detected by MTT and clone formation, and Raji cell cycle and apoptosis were detected by flow cytometry. The results showed that miR-181b can bind to bases 21–42 of the FAMLF 5′ untranslated region (UTR), FAMLF was highly expressed and miR-181b was lowly expressed in BL patients compared with unaffected individuals. FAMLF expression was significantly and inversely correlated to miR-181b expression, and miR-181b negatively regulated FAMLF at posttranscriptional and translational levels. A dual-luciferase reporter gene assay identified that the 5′ UTR of FAMLF mRNA contained putative binding sites for miR-181b. Down-regulation of FAMLF by miR-181b arrested cell cycle, inhibited cell viability and proliferation in a BL cell line model. Our findings explain a new mechanism of BL pathogenesis and may also have implications in the therapy of FAMLF-overexpressing BL.

Humans , Male , Female , Infant , Child, Preschool , Child , Adolescent , Adult , Young Adult , Burkitt Lymphoma/genetics , Burkitt Lymphoma/metabolism , Gene Expression Regulation, Neoplastic , MicroRNAs/metabolism , Proteins/metabolism , Apoptosis/genetics , Cell Line, Tumor , Cell Proliferation/genetics , Cell Survival/genetics , Down-Regulation/genetics , MicroRNAs/genetics , Proteins/genetics
Arch. latinoam. nutr ; 66(1): 74-81, mar. 2016. ilus, tab
Article in Spanish | LILACS, LIVECS | ID: biblio-1023550


El potencial de uso de la harina de calamar gigante (Dosidicus gigas) (HCG). como alternativa en el desarrollo de productos con valor agregado es relevante. Sin embargo hace falta conocer los elementos químicos que la conforman. El objetivo de este trabajo fue determinar la composición química de la harina de calamar gigante (Dosidicus gigas) procedente de Guaymas, Sonora, México y su posible alternativa para el desarrollo de alimentos funcionales. Los resultados indicaron un alto contenido de proteína (77,7%), sobresaliendo lisina y ácido glutámico (10,16 y 14,53 g aa/100g proteína respectivamente), aminoácidos azufrados y aminoácidos hidrofóbicos. El contenido de la fracción grasa (6,3%) fue bajo así como el de fibra cruda (2,7%), reportada como quitina, reflejándose en el bajo aporte calórico (4 kcal/g). La relación entre ácidos grasos saturados, monoinsaturados y poliinsaturados fue de 1,66:1:1,08 y de n6:n3 fue de 1:1,35. Se concluye que HCG es un ingrediente con posibilidades de uso en panificación, galletas saladas, sazonadores, aderezos, a los que les podría dar un valor agregado. Sin embargo el factor limitante para su uso está en el olor y sabor a pescado, por lo que su aplicación se sugiere dirigir la aplicación hacia el desarrollo de nuevos productos vinculados con preparaciones típicas que incluyan pescados y derivados(AU)

The potential use of giant squid (Dosidicus gigas) meal (GSM) as an alternative in the development of value-added foods may be relevant. However one must know the chemical elements that constitute it. The aim of this study was to determine the chemical composition of giant squid Dosidicus gigas meal from Guaymas, Sonora, Mexico and its possible alternative for the development of functional foods. The data indicated a high protein content (77,7%), lysine and glutamic acid (10,16 and 14,53 g aa/100g protein respectively), sulfur amino acids and hydrophobic amino acids. The content of fat fraction (6,3%) was low and crude fiber (2,7%) reported as chitin, reflected in the low calorie (4 kcal/g). The ratio of saturated, monounsaturated and polyunsaturated fatty acids was 1,66: 1: 1,08 and n6: n3 was 1: 1,35. It is concluded that GSM is an ingredient with potential for use in bread, crackers, seasonings, dressings, which could give them added value. However the limiting factor for use is in the smell and taste of fish, so that its application would be directed at the development of new products related with typical preparations that include fish and derivatives(AU)

Humans , Male , Female , Decapodiformes , Proteins/metabolism , Fish Flour , Food Handling/methods , Eating , Food Quality , Fatty Acids
Rev. bras. anestesiol ; 65(5): 384-394, Sept.-Oct. 2015. tab
Article in English | LILACS | ID: lil-763142


ABSTRACTBACKGROUND AND OBJECTIVES: Although many recognize that the first year of life and specifically the neonatal period are associated with increased risk of anesthetic morbidity and mortality, there are no studies directed to these pediatric subpopulations. This systematic review of the scientific literature including the last 15 years aimed to analyze the epidemiology of morbidity and mortality associated with general anesthesia and surgery in the first year of life and particularly in the neonatal (first month) period.CONTENT: The review was conducted by searching publications in Medline/PubMed databases, and the following outcomes were evaluated: early mortality in the first year of life (<1 year) and in subgroups of different vulnerability in this age group (0-30 days and 1-12 months) and the prevalence of cardiac arrest and perioperative critical/adverse events of various types in the same subgroups.CONCLUSIONS: The current literature indicates great variability in mortality and morbidity in the age group under consideration and in its subgroups. However, despite the obvious methodological heterogeneity and absence of specific studies, epidemiological profiles of morbidity and mortality related to anesthesia in children in the first year of life show higher frequency of morbidity and mortality in this age group, with the highest peaks of incidence in the neonates' anesthesia.

RESUMOJUSTIFICATIVA E OBJETIVOS: Embora muitos reconheçam que a idade inferior a um ano e especificamente o período neonatal estejam associados a maior risco de morbimortalidade anestésica, não existem estudos dirigidos a essas subpopulações pediátricas. Esta revisão sistemática das publicações científicas dos últimos 15 anos teve como objetivo analisar o perfil epidemiológico da morbimortalidade relacionada com a anestesia geral e cirurgia no primeiro ano de idade e em particular no período neonatal (primeiro mês de idade).CONTEúDO: A revisão foi conduzida por pesquisa de publicações nas bases de dados Medline/PubMed. Foram avaliados os seguintes desfechos: mortalidade precoce no primeiro ano de idade (< 1A) e em subgrupos de diferente vulnerabilidade nesta faixa etária (0-30 dias e 1-12 meses) e prevalência de parada cardíaca e eventos críticos/adversos perioperatórios de diversos tipos nos mesmos subgrupos.CONCLUSÕES: A literatura corrente indica grande variabilidade nos índices de mortalidade e morbidade na faixa etária em análise, bem como nos seus subgrupos. No entanto, apesar da óbvia heterogeneidade metodológica e da ausência de estudos específicos, os perfis epidemiológicos de morbimortalidade relacionada com a anestesia de crianças no primeiro ano de idade mostram frequência mais alta de morbimortalidade nessa faixa etária, com os maiores picos de incidência na anestesia de neonatos.

Binding Sites , Protein Binding , Protein Conformation , Proteins/chemistry , Proteins/metabolism , Computational Biology , Databases, Protein , Models, Molecular , Nuclear Magnetic Resonance, Biomolecular
Colomb. med ; 46(3): 135-143, July-Sept. 2015. ilus
Article in English | LILACS | ID: lil-765514


Objective: Our objective was to systematically review the published observational research related to the role of oxidative-nitrosative stress in pathogenesis of dengue. Methods: We searched electronic databases (PubMed, EMBASE, The COCHRANE library, ScienceDirect, Scopus, SciELO, LILACS via Virtual Health Library, Google Scholar) using the term: dengue, dengue virus, severe dengue, oxidative stress, nitrosative stress, antioxidants, oxidants, free radicals, oxidized lipid products, lipid peroxides, nitric oxide, and nitric oxide synthase. Articles were selected for review by title and abstract excluding letter, review, in vivo and in vitro studies, and duplicates studies. Selected articles were reviewed for study design, original purposes, sample size, main outcomes, methods, and oxidative-nitrosative stress markers values. Results: In total, 4,331 non-duplicates articles were identified from electronic databases searches, of which 16 were eligible for full text searching. Data from the observational studies originate from Asian countries (50%; 8/16), South American countries (31.2%; 5/16), and Central America and the Caribbean countries (18.8%; 3/16). Case-control study was the type of design most common in researches reviewed. The 1997 World Health Organization (WHO) dengue case classification criteria were used in all studies included in this review. Conclusions: Based on published data found in peer-reviewed literature, oxidative and nitrosative stress are demonstrated by changes in plasma levels of nitric oxide, antioxidants, lipid peroxidation and protein oxidation markers in patients with dengue infection. Additionally, elevated serum protein carbonyls and malondialdehyde levels appear to be associated with dengue disease severity.

Objetivo: Sistematizar las evidencias observacionales sobre la relación entre el estrés oxidativo-nitrosativo y la patogénesis del dengue. Métodos: Revisión sistemática de estudios observacionales en las bases de datos (PubMed, EMBASE, The COCHRANE library, ScienceDirect, Scopus, SciELO, LILACS via Virtual Health Library, Google Scholar) utilizando las siguientes palabras clave: dengue, dengue virus, severe dengue, oxidative stress, nitrosative stress, antioxidants, oxidants, free radicals, oxidized lipid products, lipid peroxides, nitric oxide y nitric oxide synthase. La selección inicial fue realizada a partir del título y resumen excluyéndose: cartas para editor, revisiones, estudios in vivo/in vitro y duplicados. A cada artículo seleccionado, se le revisó el diseño del estudio, objetivo, tamaño de la muestra, resultados principales y niveles plasmáticos de los marcadores de estrés oxidativo-nitrosativo. Resultados: De 4,331 publicaciones encontradas, 16 estudios cumplieron con los criterios de inclusión. El 50% (8/16) de los estudios revisados fueron realizados en países de Sur América, Centro América y del Caribe. El diseño de casos y controles fue el más frecuente. El anterior sistema de clasificación de casos (OMS-1997) fue utilizado en todos los estudios incluidos en esta revisión. Conclusiones: El estrés oxidativo-nitrosativo se encuentra presente en el curso de la infección por virus dengue, demostrado por los cambios en las concentraciones plasmáticas de óxido nítrico, antioxidantes y marcadores de lipoperoxidación y de oxidación de proteínas. Por último, parece existir una asociación entre la elevación de los niveles plasmáticos de los carbonilos proteicos y malondialdehído con la severidad del dengue.

Animals , Humans , Dengue/physiopathology , Nitric Oxide/metabolism , Oxidative Stress , Antioxidants/metabolism , Lipid Peroxidation/physiology , Proteins/metabolism , Severity of Illness Index
Electron. j. biotechnol ; 18(2): 122-127, Mar. 2015. ilus, graf, tab
Article in English | LILACS | ID: lil-745580


Background The objective of this study was to investigate proliferator-activated receptor (PPARγ), fatty acid synthase (FAS) and hormone-sensitive lipase (HSL) mRNA and protein expression in fat tails of Tan sheep. Rams from different developmental stages (aged 3, 6, 9, 12, 15 and 18 months) were selected, and their tail measurements including length (L), width (W) and girth (G) were recorded. The mRNA and protein expressions of PPARγ, FAS and HSL were examined by quantitative real-time polymerase chain reaction (PCR) and Western blot. Results The tail measurements increased with age. We observed no significant differences (P > 0.05) of PPARγ mRNA expression between ages 9 and 15 months, and between 12 and 15 months; FAS mRNA expression levels at each developmental stage were observed significantly in Tan sheep (P < 0.05); HSL mRNA expression with no significant differences were only observed between 6 and 15 months (P > 0.05). Significant differences (P < 0.05) of PPARγ, FAS and HSL protein expressions at each developmental stage were observed in Tan sheep. Conclusion We observed that the mRNA expression patterns of PPARγ and FAS decreased first before they increased again and then this process repeated. Conversely, the mRNA expression patterns of HSL increased first before they decreased and then this process repeated. The protein expression patterns of PPARγ and FAS decreased first before they increased again and then this process repeated. Conversely, the protein expression pattern of HSL increased first before it decreased again and then increased again.

Animals , Sheep/growth & development , Sheep/genetics , Proteins/metabolism , Sterol Esterase/metabolism , PPAR gamma/metabolism , Fatty Acid Synthases/metabolism , Transcription Factors , RNA, Messenger , Blotting, Western , Sterol Esterase/genetics , PPAR gamma/genetics , Fatty Acid Synthases/genetics , Real-Time Polymerase Chain Reaction
J. appl. oral sci ; 23(1): 79-86, Jan-Feb/2015. tab, graf
Article in English | LILACS, BBO | ID: lil-741593


Objective The identification of stem cells (SC) remains challenging. In the human oral mucosal epithelium, these cells are believed to be in the basal layer (stem cell niche), but their exact location is unclear. The aim of this study was to examine the dysplastic oral epithelium for these SC-like proteins in order to assess their diagnostic value as biomarkers complementing the histological grading of dysplasia. Material and Methods Thirty oral epithelial dysplasia (OED), 25 oral lichen planus (OLP), 10 oral hyperkeratosis and 5 normal oral epithelium (OE) were immunohistochemically examined for four SC markers [integrin β1, neuron-glial-2 (NG2), notch 1 (N1) and keratin 15 (K15)]. Results Three of four SC markers were heterogeneously detected in all samples. K15 overexpression in the lower two-thirds of severe OED suggests an expanded SC niche. Integrin β1 distribution pattern was not measurably different between OEDs and control. NG2 was almost negative to absent in all samples examined. N1 expression was weak and highly variable in normal and dysplastic epithelium, making it an unreliable epithelial stem cell marker. Conclusions Present findings suggest that these markers were unable to identify individual epithelial stem cells. Instead, subpopulations of cells, most probably stem cells and transit amplifying cells with stem cell-like properties were identified in the dysplastic oral epithelium. The characteristic expressions of K15 might be of diagnostic value for oral dysplasia and should be investigated further. .

Humans , Epithelial Cells/metabolism , Proteins/metabolism , Stem Cells/metabolism , /analysis , Antigens/analysis , Biomarkers/analysis , Epithelial Cells/pathology , Hyperplasia/metabolism , Immunohistochemistry , /analysis , Lichen Planus, Oral/metabolism , Lichen Planus, Oral/pathology , Mouth Mucosa/metabolism , Mouth Mucosa/pathology , Paraffin Embedding , Proteoglycans/analysis , Receptor, Notch1/analysis , Reference Values , Severity of Illness Index , Stem Cells/pathology
Article in English | WPRIM | ID: wpr-106133


BACKGROUND/AIMS: Acute exacerbations in chronic obstructive pulmonary disease may be related to air pollution, of which ozone is an important constituent. In this study, we investigated the protein profiles associated with ozone-induced exacerbations in a smoking-induced emphysema model. METHODS: Mice were divided into the following groups: group I, no smoking and no ozone (NS + NO); group II, no smoking and ozone (NS + O); group III, smoking and no ozone (S + NO); and group IV, smoking and ozone (S + O). Bronchoalveolar lavage, the mean linear intercept (MLI) on hematoxylin and eosin staining, nano-liquid chromatography-tandem mass spectrometry (LC-MS/MS), and Western blotting analyses were performed. RESULTS: The MLIs of groups III (S + NO) and IV (S + O) (45 +/- 2 and 44 +/- 3 microm, respectively) were significantly higher than those of groups I (NS + NO) and II (NS + O) (26 +/- 2 and 23 +/- 2 microm, respectively; p < 0.05). Fourteen spots that showed significantly different intensities on image analyses of two-dimensional (2D) protein electrophoresis in group I (NS + NO) were identified by LC-MS/MS. The levels of six proteins were higher in group IV (S + O). The levels of vimentin, lactate dehydrogenase A, and triose phosphate isomerase were decreased by both smoking and ozone treatment in Western blotting and proteomic analyses. In contrast, TBC1 domain family 5 (TBC1D5) and lamin A were increased by both smoking and ozone treatment. CONCLUSIONS: TBC1D5 could be a biomarker of ozone-induced lung injury in emphysema.

Animals , Male , Biomarkers/metabolism , Blotting, Western , Bronchoalveolar Lavage Fluid/chemistry , Chromatography, Liquid , Disease Models, Animal , Electrophoresis, Gel, Two-Dimensional , Lung/metabolism , Mice, Inbred C57BL , Ozone , Proteins/metabolism , Proteomics/methods , Pulmonary Disease, Chronic Obstructive/etiology , Pulmonary Emphysema/etiology , Smoking/adverse effects , Tandem Mass Spectrometry
Article in English | WPRIM | ID: wpr-164158


This study is a multi-center clinical study, which aimed to compare CA125, HE4, and risk of ovarian malignancy algorithm (ROMA) in predicting epithelial ovarian cancer of Korean women with a pelvic mass. Prospectively, serum from 90 Korean women with ovarian mass was obtained prior to surgery. For control group, serum from 79 normal populations without ovarian mass was also obtained. The HE4 and CA125 data were registered and evaluated separately and ROMA was calculated for each sample. Total 67 benign tumors and 23 ovarian cancers were evaluated. Median serum levels of HE4 and CA125, and ROMA score were significantly higher in patients with ovarian cancer than those with benign ovarian tumor and normal population (P < 0.001). In ROC curve analysis for women with a pelvic mass, area under the curve (AUC) for HE4 and ROMA was higher than CA125. Statistical differences in each study compared to CA125 were marginal (P compared to CA125; 0.082 for HE4 and 0.069 for ROMA). Sub-analysis revealed that AUC for HE4 and ROMA was higher than AUC for CA125 in post-menopausal women with a pelvic mass, but there were no statistically significant differences (P compared to CA125; 0.160 for HE4 and 0.127 for ROMA). Our data suggested that both HE4 and ROMA score showed better performance than CA125 for the detection of ovarian cancer in women with a pelvic mass. HE4 and ROMA can be a useful independent diagnostic marker for epithelial ovarian cancer in Korean women.

Female , Humans , Middle Aged , Algorithms , Area Under Curve , Biomarkers, Tumor/blood , CA-125 Antigen/blood , Case-Control Studies , Neoplasms, Glandular and Epithelial/blood , Ovarian Neoplasms/blood , Predictive Value of Tests , Prospective Studies , Proteins/metabolism , ROC Curve , Reference Values , Republic of Korea