ABSTRACT
BACKGROUND: Salinity is a serious factor limiting the productivity of agricultural plants. One of the potential problems for plants growing under saline conditions is the inability to up take enough K+. The addition of K+ may considerably improve the salt tolerance of plants grown under salinity. It is assumed that increasing the K+ supply at the root zone can ameliorate the reduction in growth imposed by high salinity. The present study aims to determine whether an increase in the K/Na ratio in the external media would enhance the growth of date palm seedlings under in vitro saline conditions. METHODS: Date palm plants were grown at four concentrations of Na + K/Cl (mol/m³) with three different K/Na ratios. The 12 salt treatments were added to modified MS medium. The modified MS medium was further supplemented with sucrose at 30 g/l. RESULTS: Growth decreased substantially with increasing salinity. Growth expressed as shoot and root weight, enhanced significantly with certain K/Na ratios, and higher weight was maintained in the presence of equal K and Na. It is the leaf length, leaf thickness and root thickness that had significant contribution on total dry weight. Na+ contents in leaf and root increased significantly increased with increasing salinity but substantial decreases in Na+ contents were observed in the leaf and root with certain K/Na ratios. This could be attributed to the presence of a high K+ concentration in the media. The internal Na+ concentration was higher in the roots in all treatments, which might indicate a mechanism excluding Na+ from the leaves and its retention in the roots. K/Na ratios up to one significantly increased the leaf and root K+ concentration, and it was most pronounced in leaves. The K+ contents in leaf and root was not proportional to the K+ increase in the media, showing a high affinity for K+ uptake at lower external K+ concentrations, but this mechanism continues to operate even with high external Na+ concentrations. CONCLUSION: Increasing K/Na ratios in the growing media of date plam significantly reduced the absorption of Na+ less than 200 mM and also balance ions compartmentalization.
Subject(s)
Potassium/metabolism , Sodium/metabolism , Crops, Agricultural , Salinity , Phoeniceae/physiology , Sucrose/pharmacology , In Vitro Techniques , Cell Compartmentation/physiology , Plant Shoots/metabolism , Plant Roots/metabolism , Plant Leaves/metabolism , Phoeniceae/growth & development , Absorption, PhysicochemicalABSTRACT
OBJECTIVES: to identify adaptation problems under Roy's Model in patients undergoing hemodialysis and to correlate them with the socioeconomic and clinical aspects. METHOD: a transversal study, undertaken using a questionnaire. The sample was made up of 178 individuals. The Chi-squared and Mann-Whitney U tests were undertaken. RESULTS: the adaptation problems and the socioeconomic and clinical aspects which presented statistical associations were: Hyperkalemia and age; Edema and income; Impairment of a primary sense: touch and income; Role failure and age; Sexual dysfunction and marital status and sex; Impairment of a primary sense: vision and years of education; Intolerance to activity and years of education; Chronic pain and sex and years of education; Impaired skin integrity and age: Hypocalcemia and access; Potential for injury and age and years of education; Nutrition below the organism's requirements and age; Impairment of a primary sense: hearing and sex and kinetic evaluation of urea; Mobility in gait and/or coordination restricted, and months of hemodialysis; and, Loss of ability for self-care, and months of hemodialysis and months of illness. CONCLUSION: adaptation problems in the clientele undergoing hemodialysis can be influenced by socioeconomic/clinical data. These findings contribute to the development of the profession, fostering the nurse's reflection regarding the care. .
OBJETIVOS: identificar os problemas adaptativos de Roy em pacientes submetidos a hemodiálise e correlacioná-los aos aspectos socioeconômicos e clínicos. MÉTODO: estudo transversal, realizado através de um formulário. A amostra foi de 178 indivíduos. Efetuaram-se os testes qui-quadrado e U de Mann-Whitney. RESULTADOS: os problemas adaptativos e os aspectos socioeconômicos e clínicos que apresentaram associações estatísticas foram: hipercalemia e idade; edema e renda; deficiência de um sentido primário: tátil e renda; falha no papel e idade; disfunção sexual e estado civil e sexo; deficiência de um sentido primário: visão e anos de estudo; intolerância à atividade e anos de estudo; dor crônica e sexo e anos de estudo; integridade da pele prejudicada e idade; hipocalcemia e acesso; potencial para lesão e idade e anos de estudo; nutrição menor que as necessidades do organismo e idade; deficiência de um sentido primário: audição e sexo e avaliação cinética da ureia; mobilidade andar e/ou coordenação restritas e meses de hemodiálise e perda de habilidade de autocuidado e meses de hemodiálise e meses de doença. CONCLUSÃO: problemas adaptativos da clientela hemodialítica podem sofrer influências de dados socioeconômicos/clínicos. Tais achados contribuem para o desenvolvimento da profissão, proporcionando reflexão por parte do enfermeiro acerca do cuidado. .
OBJETIVOS: identificar los problemas adaptativos de Roy en pacientes sometidos a hemodiálisis y correlacionarlos a los aspectos socioeconómicos y clínicos. MÉTODO: estudio transversal, realizado a través de un formulario. La muestra fue de 178 individuos. Se efectuaron las pruebas Chi-cuadrado y U de Mann-Whitney. RESULTADOS: los problemas adaptativos y los aspectos socioeconómicos y clínicos que presentaron asociaciones estadísticas fueron: Hiperkalemia y edad; Edema y renta; Deficiencia de un sentido primario: táctil y renta; Fracaso en el papel y edad; Disfunción sexual y estado civil y sexo; Deficiencia de un sentido primario: visión y años de estudio; Intolerancia a la actividad y años de estudio; Dolor crónico y sexo y años de estudio; Integridad de la piel perjudicada y edad; Hipocalcemia y acceso; Potencial para lesión y edad y años de estudio; Nutrición menor que las necesidades del organismo y edad; Deficiencia de un sentido primario: audición y sexo y evaluación cinética de la urea; Movilidad andar y/o coordinación restringidas y meses de hemodiálisis; y, Pérdida de habilidad de autocuidado y meses de hemodiálisis y meses de enfermedad. CONCLUSIÓN: los problemas adaptativos de la clientela hemodialítica pueden sufrir influencias de datos socioeconómicos/clínicos. Esos hallazgos contribuyen para el desarrollo de la profesión, permitiendo la reflexión del enfermero acerca del cuidado. .
Subject(s)
Humans , Lysosomes/metabolism , Proteins/metabolism , Ubiquitins/physiology , Cell Compartmentation , Cysteine Proteinase Inhibitors/pharmacology , Intermediate Filaments/physiology , Leucine/analogs & derivatives , Leucine/pharmacology , Metabolism, Inborn Errors/metabolism , Organelles/ultrastructureABSTRACT
OBJECTIVE: to relate neck circumference with metabolic syndrome and its criteria among college students. METHOD: cross-sectional study conducted with 702 college students in Fortaleza, CE, Brazil from September 2010 to June 2011. Socio-demographic data, waist circumference and neck circumference were collected together with blood pressure, fasting blood sugar, triglyceride levels, and HDL-C. RESULTS: 1.7% of the studied sample presented metabolic syndrome. Of these, 58.3% presented altered neck circumference (p<0.006). As neck circumference decreases, pressure levels improve (p<0.001). Additionally, college students with high fasting blood sugar (p=0.003) and high triglyceride levels (p<0.001) presented higher values of neck circumference. CONCLUSION: neck circumference is a potential predictive marker in the detection of metabolic syndrome and its components among college students. .
OBJETIVO: relacionar a circunferência do pescoço com a síndrome metabólica e seus critérios em universitários. MÉTODO: estudo transversal, realizado com 702 universitários de Fortaleza, CE, Brasil, no período de setembro de 2010 a junho de 2011. Coletaram-se dados sociodemográficos, circunferência da cintura, circunferência do pescoço, níveis de pressão arterial e glicemia plasmática de jejum, triglicerídeos e lipoproteína de alta densidade. RESULTADOS: 1,7% da amostra investigada tinha a síndrome metabólica. Desses, 58,3% apresentaram circunferência do pescoço alterada (p<0,006). Na medida em que decresce a circunferência do pescoço, os valores pressóricos dos universitários melhoram (p<0,001). Também, observou-se que universitários com valores de glicemia de jejum plasmática (p=0,003) e triglicerídeos (p<0,001) elevados apresentaram maiores valores de circunferência do pescoço. CONCLUSÃO: a circunferência do pescoço mostrou-se um possível marcador preditivo para detecção da síndrome metabólica e seus componentes em universitários. .
OBJETIVO: relacionar la circunferencia del cuello con el síndrome metabólico y sus criterios en universitarios. MÉTODO: estudio transversal realizado con 702 universitarios de Fortaleza-CE, Brasil, en el período de septiembre de 2010 a junio de 2011. Se recolectaron datos sociodemográficos, circunferencia de la cintura, circunferencia del cuello, niveles de presión arterial y glucemia plasmática de ayuno, triglicéridos y HDL-C. RESULTADOS: 1,7% de la muestra investigada tenían el síndrome metabólico. De estos, 58,3% presentaron circunferencia del cuello alterada (p<0,006). A medida que decrece la circunferencia del cuello mejoran los valores de la presión de los universitarios (p<0,001). También, se observó que los universitarios con valores de glucemia de ayuno plasmática (p=0,003) y triglicéridos (p<0,001) elevados presentaron mayores valores de circunferencia del cuello. CONCLUSIÓN: la circunferencia del cuello se mostró un posible indicador de predicción para la detección del síndrome metabólico y sus componentes, en universitarios. .
Subject(s)
Humans , Animals , Cathepsins/physiology , Lysosomes/metabolism , Proteins/metabolism , Amino Acid Sequence , Autophagy , Base Sequence , Cathepsins/antagonists & inhibitors , Cathepsins/genetics , Cell Compartmentation , Cycloheximide/pharmacology , Cystatins/physiology , Gene Expression Regulation , Leucine/analogs & derivatives , Leucine/pharmacology , Lysosomes/enzymology , Molecular Sequence Data , Muscular Diseases/physiopathology , Restriction MappingABSTRACT
Objective To understand the experiences and expectations of nurses in the treatment of women with chronic venous ulcers. Method Phenomenological research was based on Alfred Schütz, whose statements were obtained in January, 2012, through semi-structured interviews with seven nurses. Results The nurse reveals the difficulties presented by the woman in performing self-care, the perceived limitations in the treatment anchored in motivation, and the values and beliefs of women. It showed professional frustration because venous leg ulcer recurrence, lack of inputs, interdisciplinary work and training of nursing staff. There was an expected adherence to the treatment of women, and it emphasized the need for ongoing care, supported self-care and standard practices in treatment. Conclusion That treatment of chronic venous leg ulcers constitutes a challenge that requires collective investment, involving women, professionals, managers and health institutions. .
Objetivo Comprender las experiencias y expectativas de enfermeras en el tratamiento de mujeres con úlcera venosa crónica. Método Investigación fenomenológica fundamentada en Alfred Schutz, que buscó Se realizó entrevista semiestructurada con siete enfermeras, en enero del 2012. Resultados La enfermera revela dificultades presentadas por la mujer para realizar el autocuidado, percibe limitaciones en el tratamiento relacionadas con la desmotivación, los valores y las creencias de las mujeres. Refiere frustración profesional debido a la recidiva de la lesión, a la falta de insumos, al deficiente trabajo interdisciplinar y a la limitada capacitación del equipo de enfermeras. Espera la adhesión de la mujer al tratamiento y resalta la necesidad del cuidado continuo, del autocuidado apoyado y de estandarizar conductas de tratamiento. Conclusión El tratamiento de la úlcera venosa crónica es un desafío que requiere contribución colectiva, involucrando a las mujeres, a los profesionales, a los gestores y a las instituciones de salud. .
Objetivo Compreender as experiências e expectativas de enfermeiras no tratamento de mulheres com úlcera venosa crônica na Atenção Primária à Saúde. Método Pesquisa fundamentada na fenomenologia social de Alfred Schütz, com depoimentos obtidos em janeiro de 2012, por meio de entrevista semiestruturada com sete enfermeiras. Resultados As enfermeiras revelam dificuldades apresentadas pelas mulheres com úlcera venosa crônica para realizar o autocuidado, percebem limitações na terapêutica ancoradas na desmotivação e nos valores e crenças das mulheres. Referem frustração profissional em razão da recidiva da lesão, falta de insumos e tecnologia, de trabalho interdisciplinar e da capacitação da equipe de enfermagem. Esperam a adesão das mulheres ao tratamento e ressaltam a necessidade do cuidado contínuo, do autocuidado apoiado e da padronização de condutas no tratamento. Conclusão O tratamento da úlcera venosa crônica constitui-se em um desafio que requer investimento coletivo, envolvendo a mulher, os profissionais, os gestores e as instituições de saúde. .
Subject(s)
Animals , Caenorhabditis elegans Proteins/isolation & purification , Caenorhabditis elegans/metabolism , Cell Membrane/metabolism , Ion Channels/isolation & purification , Ion Channels/metabolism , Nerve Tissue Proteins/isolation & purification , Nerve Tissue Proteins/metabolism , Nervous System/metabolism , Neurons, Afferent/metabolism , Sensation/genetics , Amino Acid Sequence/genetics , Base Sequence/genetics , Behavior, Animal/drug effects , Behavior, Animal/physiology , Caenorhabditis elegans Proteins/genetics , Caenorhabditis elegans Proteins/metabolism , Caenorhabditis elegans/cytology , Capsaicin/pharmacology , Cell Compartmentation/genetics , Cell Membrane/drug effects , Cell Membrane/ultrastructure , Gene Expression Regulation/physiology , Ion Channels/genetics , Ion Channels/ultrastructure , Molecular Sequence Data , Mutation/genetics , Nerve Tissue Proteins/genetics , Nerve Tissue Proteins/ultrastructure , Nervous System/cytology , Nervous System/drug effects , Neurons, Afferent/cytology , Neurons, Afferent/drug effects , Pain/genetics , Pain/metabolism , Pain/physiopathology , Phylogeny , Receptors, Drug/drug effects , Receptors, Drug/metabolism , Receptors, Drug/ultrastructure , Sensation/drug effects , Signal Transduction/genetics , TRPV Cation Channels , Transient Receptor Potential ChannelsABSTRACT
This experimental research was aimed to establish an optimum system of enucleation, purification and identification for preparing the cytoplasts of suspension culture cells in order to undertake cell recombination. Human leukemia HL-60 cells in suspension culture were purified by 42% Percoll density gradient centrifugation and low-speed centrifugation at 1 500r/min, respectively. The purified HL-60 cells were treated with cytochalasin B (CB) alone or combined with colcchicine and enucleated by isopycnic gradient centrifugation on 50% Percoll at 25 degrees C and 34 degrees C, respectively. Cytoplasts made from HL-60 cells were purified through gradient centrifugation by 37%, 38% and 40% Percoll, respectively. The final cytoplasts were identified by Wright-Giemsa staining and 4,6-diamidino-2-phenylidole dihydrochloride (DAPI)/5, 6-carboxyflu-orescein diacetate succinimidyl ester (CFSE) double-staining. The phenotype and mitochondrial membrane potential of HL-60 cytoplasts were analyzed by flow cytometry. The results indicated that the enucleation ratio of HL-60 cells induced by CB combined with colcchicine was up to 91. 98% +/-4. 29%, which was significantly higher than that in CB alone group (74. 95% +/- 3. 02%)(P<0. 01). The rates of enucleation and cytoplast with diameter over 5min in 34 degrees C group were higher than those in 25 degrees C group (all P<0. 01). The cytoplast purities were (95.43 +/- 0. 59)% in 38% Percoll groups,which were higher than those of 40% Percoll (P<0. 05). Nucleus and caryoplasm could be clearly distinguished by DAPI and CFSE double labeling. The results further showed that the phenotype of HL-60 cytoplasts had no significant change, and the activity of the cytoplasts was above 80% within 12h. It is concluded that enucleation throuth density gradient centrifugation on 50% Percoll mediated by CB combined with colcchicine, 38%Percoll of purification followed by DAPI/CFSE double labeling and MMP detection is an optimum scheme for preparation and identification of cytoplast from suspension culture cells.
Subject(s)
Humans , Cell Compartmentation , Cell Nucleus , Cell Separation , Centrifugation, Density Gradient , Colchicine , Pharmacology , Cytochalasin B , Pharmacology , Cytoplasm , HL-60 CellsABSTRACT
El microquimerismo humano se define como la presencia dentro de un individuo de una pequeña población de células originadas en un individuo genéticamente distinto. El microquimerismo adquirido de forma natural se desarrolla a lo largo del embarazo, como una consecuencia del ampliamente demostrado tráfico bidireccional transplacentario de células. Esta trasferencia resulta en un estado microquimérico a décadas posparto, tanto para la madre (microquimerismo fetal) como para su progenie (microquimerismo materno). Aún más, el establecimiento de esta condición ocurre, iatrogénicamente, posterior al transplante de órganos y la transfusión de sangre. Actualmente el microquimerismo es un tema de gran interés alrededor del mundo, dadas sus múltiples posibles implicaciones en la salud y en la enfermedad. Su estudio comenzó a raíz del potencial para el desarrollo de técnicas de diagnóstico prenatal no invasivo, en le presente se tienen amplias expectativas de su utilidad en la terapia con células madre. Se ha propuesto que el microquimerismo podría estar implicado en la patogénesis de ciertas enfermedades autoinmunes, en la preparación de tejidos, el cáncer y la inducción de tolerancia...
Subject(s)
Humans , Autoimmune Diseases , Cell Compartmentation , Cell Movement , Genome, Human , Placenta , Prenatal Diagnosis , Stem Cells , Costa RicaABSTRACT
Unipolar brush cells (UBCs) are excitatory interneurons with their somata located in the granular layer. Recently, T-brain factor 2 (Tbr2) was shown to be expressed in a subset of UBCs in mouse cerebellum. Scrambler mice exhibit severe cerebellum abnormalities, including the failure of embryonic Purkinje cell dispersal and a complete absence of foliation due to a mutation in the disabled-1 adaptor protein. Since most UBC markers are expressed postnatally, it has proven difficult to identify the relationship between developing Purkinje cell clusters and migrating UBCs. Because scrambler mice closely mimic normal embryonic day 18 cerebellum, we examined whether Tbr2-positive UBCs are associated with Purkinje cell cluster markers such as zebrin II, which is the most studied compartmentation marker in the cerebellum. We investigated the distribution of Tbr2-positive UBCs in this mutant by using anti-Tbr2 immunocytochemistry. The data revealed that Tbr2 immunoreactivity was exclusively present in the nucleus of UBCs in scrambler cerebellum. Based on expression data, a Tbr2-positive UBC map was constructed. In addition, Tbr2-positive UBCs are found associated with ectopic zebrin II-immunoreactive Purkinje cell clusters in scrambler cerebellum. These data suggest that UBCs use Purkinje cell compartmentation to migrate into their final position through interactions with the embryonic array of specific Purkinje cell subtypes.
Subject(s)
Animals , Mice , Cell Compartmentation , Cerebellum , Hydrazines , Immunohistochemistry , Interneurons , Nerve Tissue ProteinsABSTRACT
Pre-mRNA splicing factors are enriched in nuclear domains termed interchromatin granule clusters or nuclear speckles. During mitosis, nuclear speckles are disassembled by metaphase and reassembled in telophase in structures termed mitotic interchromatin granules (MIGs). We analysed the dynamics of the splicing factor SC35 in interphase and mitotic cells. In HeLa cells expressing green fluorescent protein (GFP)-SC35, this was localized in speckles during interphase and dispersed in metaphase. In telophase, GFP-SC35 was highly enriched within telophase nuclei and also detected in MIGs. Fluorescence recovery after photobleaching (FRAP) experiments revealed that the mobility of GFP-SC35 was distinct in different mitotic compartments. Interestingly, the mobility of GFP-SC35 was 3-fold higher in the cytoplasm of metaphase cells compared with interphase speckles, the nucleoplasm or MIGs. Treatment of cells with inhibitors of cyclin-dependent kinases (cdks) caused changes in the organization of nuclear compartments such as nuclear speckles and nucleoli, with corresponding changes in the mobility of GFP-SC35 and GFP-fibrillarin. Our results suggest that the dynamics of SC35 are significantly influenced by the organization of the compartment in which it is localized during the cell cycle.
Subject(s)
Cell Compartmentation , Fluorescence Recovery After Photobleaching , Green Fluorescent Proteins/genetics , HeLa Cells , Humans , Interphase , Mitosis , Nuclear Proteins/genetics , Ribonucleoproteins/geneticsABSTRACT
Determinar o risco da ocorrência de metástase do carcinoma bem diferenciado da glândula tireóide auxilia na escolha do tratamento adequado. Um modelo de predição individual do risco de metástase foi desenvolvido com base no estudo caso-controle de um grupo de 109 doentes operados e seguidos por mais de 5 anos. A imunomarcação da galectina-3 no nucléolo do tireócito maligno foi um fator preditivo do risco individual de metástase linfática cervical...
Determination of metastasis risk in well differentiated thyroid carcinoma allows surgeons and oncologists to offer an adequate treatment to each patient. Individual metastasis risk prediction was calculated based on a case-control study involving 109 treated patients who were followed up during at least 5 years. Nucleolar galectin-3 immuno expression in malignant thyrocytes was an individual predictive factor for cervical lymph node metastasis...
Subject(s)
Male , Female , Humans , Adenocarcinoma, Papillary , Carcinoma, Papillary , Thyroid Neoplasms , Cell Compartmentation , /analysis , Immunohistochemistry , Logistic Models , Biomarkers, Tumor , Neoplasm MetastasisABSTRACT
<p><b>BACKGROUND</b>The aim of this study was to identify the subnuclear distribution pattern of human orphan nuclear receptor steroidogenic factor 1 (SF-1) in living cells with and without the activation of protein kinase A (PKA) signal pathway, and thus try to explain the unknown mechanism by which PKA potentiates SF-1 transactivation.</p><p><b>METHODS</b>Full-length cDNAs of wild type and a naturally occurring mutant (G35E) human SF-1 were cloned and fused with green fluorescent protein (GFP). Subcellular distribution pattern of human SF-1 in living cells, whose PKA signaling was either activated or not, was studied by laser confocal microscopy after the validity of the gene sequence was confirmed.</p><p><b>RESULTS</b>The transactivation ability of the GFP-SF-1 chimeric protein was highly conserved. Wild type human SF-1 diffused homogeneously within the nuclei of cells when PKA was not active, and converged to clear foci when PKA was activated. Mutant SF-1 diffused within the nuclei even in the presence of PKA activation, surprisingly aggregating as fluorescent dots inside the nucleoli, a phenomenon not altered by PKA.</p><p><b>CONCLUSIONS</b>Activation of PKA causes wild type, but not mutant SF-1 to alter its subnuclear distribution pattern to a transactivationally active form (foci formation). This finding may throw new light on the mechanism by which PKA activates the orphan nuclear receptor.</p>
Subject(s)
Female , Humans , Cell Compartmentation , Cell Nucleus , Chemistry , Cells, Cultured , Colforsin , Pharmacology , Cyclic AMP-Dependent Protein Kinases , Physiology , DNA-Binding Proteins , Enzyme Activation , Fushi Tarazu Transcription Factors , Homeodomain Proteins , Microscopy, Confocal , Receptors, Cytoplasmic and Nuclear , Steroidogenic Factor 1 , Transcription Factors , Transcriptional ActivationABSTRACT
Protein domains play a fundamental role in the spatial and temporal organization of intracellular signaling systems. While protein phosphorylation has long been known to modify the interactions that underlie this organization, the dynamic cycling of lipids should now be included amongst the posttranslational processes determining specificity in signal transduction. The characteristics of this process are reminiscent of the properties of protein and lipid phosphorylation in determining compartmentalization through SH2 or PH domains. Recent studies have confirmed the functional importance of protein S-palmitoylation in the compartmentalization of signaling molecules that support normal physiological function in cell division and apoptosis, and synaptic transmission and neurite outgrowth. In neurons, S-palmitoylation and targeting of proteins to rafts are regulated differentially in development by a number of processes, including some related to synaptogenesis and synaptic plasticity. Alterations in the S-palmitoylation state of proteins substantially affect their cellular function, raising the possibility of new therapeutic targets in cancer and nervous system injury and disease
Subject(s)
Humans , Animals , Rats , Cell Compartmentation , Neurons , Palmitates , Protein Processing, Post-Translational , Protein S , Signal Transduction , Growth Cones , Protein S , SynaptosomesABSTRACT
Cytosolic calcium signals are produced by suddenly increasing the concentration of free calcium ions (Ca2+). This can occur by opening channels permeable to Ca2+ either in the surface cell membrane or in the membranes of intracellular organelles containing high Ca2+ concentrations. Ca2+ signals can control several different processes, even in the same cell. In pancreatic acinar cells, for example, Ca2+ signals do not only control the normal secretion of digestive enzymes, but can also activate autodigestion and programmed cell death. Recent technical advances have shown that different patterns of Ca2+ signals can be created, in space and time, which allow specific cellular responses to be elicited. The mechanisms responsible for Ca2+ signal compartmentalization are now largely known and will be described on the basis of recent studies of Ca2+ transport pathways and their regulation in pancreatic acinar cells. It turns out that the Ca2+ handling as well as the structural characteristics of the endoplasmic reticulum (ER) and the mitochondria are of particular importance. Using a variety of Ca(2+)-sensitive fluorescent probes placed in different sub-cellular compartments in combination with local uncaging of caged Ca2+, many new insights into Ca2+ signal generation, compartmentalization and termination have recently been obtained
Subject(s)
Humans , Animals , Calcium Signaling , Cell Compartmentation , Pancreas , Calcium Channels , Cell DeathABSTRACT
The localization and subcellular distribution of Trypanosoma cruzi nitric oxide synthase was investigated in epimastigote cells by immunocytochemistry at electron and light microscope level, using a polyclonal antibody to neuronal nitric oxide synthase, and also, at light microscope level, by the nicotinamide adenine dinucleotide phosphate-diaphorase histochemical reaction. The immunoreactivity was ultrastructurally localized by electron microscopy in the inner surface of cell membranes and in free cytosolic clusters in the body, flagellum and apical extreme. Light microscopy showed that immunoprecipitates, specific for the Trypanosoma cruzi nitric oxide synthase, co-localized with the formazan precipitates generated by the diaphorase reaction in the same areas identified by electron microscopy. These results, taken together with previous finding from our laboratory could help to explain the involvement of the nitric oxide transduction pathway in T. cruzi epimastigote motility.
Subject(s)
Animals , Cell Movement , Nitric Oxide Synthase , Nitric Oxide/metabolism , Signal Transduction/physiology , Trypanosoma cruzi , Cell Compartmentation , Cell Membrane , FlagellaABSTRACT
The objective of this review is to examine the biogenesis of lysosomes during the endocytic flow of plasma membrane. Two models have been proposed to explain the formation of lysosomes by this process: the "maturational" and the "stationary" models. According to the former, pinocytotic vesicles fuse among themselves to yield endosomes, which in turn, transform first into multivesicular bodies (MVB) and then into mature lysosomes. Therefore, endosomes and lysosomes would be transient organelles. On the other hand, the "stationary" model proposes that the endocytic pathway is formed by functionally and physically distinct compartments. Cultured cells exposed to ammonium chloride (NH4Cl) and leupeptin after a pulse of cationic ferritin were recently used to freeze endosomes and lysosomes. NH4Cl produced a retention of cationic ferritin in endosomes, indicating that this agent interfered with the endosomal/lysosomal progression. Leupeptin did not affect this process. The number of lysosomes increased in cells treated with both lysosomotropic agents. Thus, NH4Cl affected the endosomal and lysosomal compartments, whereas leupeptin had a preferential effect on lysosomes. Mice mutants with defects of plasma membrane degradation, including a Tay-Sachs model, a Sandhoff disease model, as well as, mice with the inactivated prosaposin gene were used to analyze the biogenesis of lysosomes. Thin sections of mutant cells were examined under the electron microscope, and the analysis revealed a selective accumulation of MVBs and the disappearance of lysosomes, suggesting that the formation of MVBs is a required step in lysosomal maturation and that the intravesicular content of MVBs is necessary for the digestion of plasma membrane components. Taken together, these data indicate that endosomes and MVBs are preceding steps in lysosomal biogenesis and that endosomes, MVBs, and lysosomes are transient organelles
Subject(s)
Animals , Cell Compartmentation , Cell Membrane , Endocytosis , Fibroblasts , LysosomesABSTRACT
Background. The mechanisms whereby rat hepatocytes undergo irreversible injury due to a lack of oxygen have not been established. Methods. Liver cells were used for reperfusion injury, and four compartmentalized pathways were evaluated durgin hypoxia (N2/CO2, 19:1) for 30 min followed by oxygen (O2/CO2, 19:1) for 30 min. Results. Cell viability decreased during the hypoxic, but not during the reoxygenation, phase. Glycogenolysis, as measured by glucose release, was significantly increased during hypoxia as compared to control in oxygen (205ñ15 vs. 155 ñ 10 mmol glucose/mg protein/h, respectiely), and did not return to normal levels by reoxygenation. Gluconeogenesis was importantly decreased during hypoxia (102 ñ 10 vs. 8 ñ 2 mmol glucose/mg protein/h) with partial recovery during reoxygenation. Ureagenesis diminished in hypoxia, but recovered during reoxygenation. Additionally, 3-hydroxy-butyrate formation was augmented by hypoxia, with some recovery when oxygen was present. Conclusions. These results suggest that compartmentalized pathways are protected from hypoxic injury in isolated hepatocytes, and also suggest it as a model to test the idea that enzymes of those parthways are organized into multienzyme complexes in vivo
Subject(s)
Animals , Male , Rats , Cell Compartmentation , Cell Hypoxia , Liver Glycogen/metabolism , Gluconeogenesis , Oxygen Consumption , Rats, Wistar , Urea/metabolismABSTRACT
Nesta revisäo säo abordados conceitos básicos sobre morte e necrose celular. Säo descritos sequencialmente as alteraçöes morfológicas e as correspondentes repercussöes bioquímicas e funcionais da agressäo celular. Nos mecanismos de morte celular enfatiza-se a açäo do cálcio, as lesöes das membranas e filamentos intermediários, os radicais livres, o papel do Ferro e as lesöes produzidas por vírus