Técnicas de Fisioterapia Respiratoria Manuales de Higiene Bronquial / Respiratory Physiotherapy Techniques Bronchial Hygiene Manuals

Este documento se enfoca en las técnicas manuales de fisioterapia respiratoria como una estrategia fundamental para la limpieza efectiva de las vías aéreas en pacientes con afecciones respiratorias. La fisioterapia respiratoria se ha establecido como un pilar crucial en la atención de individuos con trastornos pulmonares obstructivos y restrictivos, así como en situaciones clínicas que requieren la eliminación de secreciones pulmonares. Existe una variedad de técnicas manuales disponibles, como la percusión torácica, la vibración, el drenaje postural y la maniobra de tos asistida, con diferentes mecanismos de acción y fundamentos fisiológicos de su aplicación. Es importante considerar la personalización de estas técnicas según las necesidades individuales de cada paciente para brindar. Un tratamiento seguro y eficaz. En el siguiente artículo se revisarán los beneficios demostrados de la fisioterapia respiratoria en términos de mejora de la función pulmonar, la oxigenación y la calidad de vida de los pacientes. Cabe destacar la importancia de la fisioterapia respiratoria como un componente esencial en el manejo de las afecciones respiratorias y destacar la necesidad de continua de investigación y desarrollo de mejores prácticas en este campo, con el objetivo de mejorar la atención y los resultados clínicos en pacientes con problemas respiratorios.

This document focuses on manual respiratory physiotherapy techniques as a fundamental strategy for effective airway clearance in patients with respiratory conditions. Respiratory physiotherapy has established itself as a crucial pillar in the care of individuals with obstructive and restrictive pulmonary disorders, as well as in clinical situations that require the removal of pulmonary secretions. There are a variety of manual techniques available, such as chest percussion, vibration, postural drainage, and the assisted cough maneuver, with different mechanisms of action and physiological rationales for their application. It is important to consider customizing these techniques to the individual needs of each patient to provide. A safe and effective treatment. The following article will review the proven benefits of respiratory physiotherapy in terms of improving lung function, oxygenation and quality of life of patients. It is worth highlighting the importance of respiratory physiotherapy as an essential component in the management of respiratory conditions and highlighting the need for continuous research and development of best practices in this field, with the aim of improving care and clinical outcomes in patients with respiratory problems.

Advances of antineoplastic agents based on mitochondrial targeting mechanism / 药学学报

Mitochondrion is an important organelle in cells and plays a crucial role in tumor development. Therefore, improvement of the targeting ability of anticancer drugs to mitochondria will increase the treatment efficacy for tumor and reduce the side effect on normal tissues. Here, research progresses in mitochondrial targeting have been reviewed in three aspects for tumors treatment: the potential, permeability and translocase of mitochondrial membrane. The review provides a reference for the development of mitochondria targeted therapeutic systems.

trans-4,4'-Dihydroxystilbene (DHS) protects PC12 cells from oxidative damage but induces reactive oxygen species-mediated apoptosis in SHSY-5Y cell line.

Polyphenols can exert both, antioxidant and pro-oxidant properties, depending on cell types as well as their concentrations. Hence, it was of interest to examine if the naturally occurring resveratrol analog, trans-4,4'-dihydroxystilbene (DHS) also exert both these activities in a biphasic or cell-specific manner. In this study, we established the cytoprotective action of DHS against hydrogen peroxide (H2O2)-induced apoptotic death of the PC12 cells. DHS reduced mitochondrial membrane permeabilization and deactivated reactive oxygen species (ROS)-mediated caspase-3 activation in the H2O2-treated PC12 cells. However, it induced apoptosis in the human neuroblastoma SHSY-5Y cell line by destabilizing mitochondrial membrane, augmenting ROS and activating caspapse-3. DHS showed better activity than resveratrol in both the chosen models.

Statistical investigation of Kluyveromyces lactis cells permeabilization with ethanol by response surface methodology

The aim of our study was to select the optimal operating conditions to permeabilize Kluyveromyces lactis cells using ethanol as a solvent as an alternative to cell disruption and extraction. Cell permeabilization was carried out by a non-mechanical method consisting of chemical treatment with ethanol, and the results were expressed as β-galactosidase activity. Experiments were conducted under different conditions of ethanol concentration, treatment time and temperature according to a central composite rotatable design (CCRD), and the collected results were then worked out by response surface methodology (RSM). Cell permeabilization was improved by an increase in ethanol concentration and simultaneous decreases in the incubation temperature and treatment time. Such an approach allowed us to identify an optimal range of the independent variables within which the β-galactosidase activity was optimized. A maximum permeabilization of 2,816 mmol L-1 oNP min-1 g-1 was obtained by treating cells with 75.0% v/v of ethanol at 20.0 °C for 15.0 min. The proposed methodology resulted to be effective and suited for K. lactis cells permeabilization at a lab-scale and promises to be of possible interest for future applications mainly in the food industry.

Iso-caryophyllene cytotoxicity induced by lipid peroxidation and membrane permeabilization in L-929 cells.

Iso-caryophyllene, a sesquiterpene, is present in several essential oils from vegetable species. In previous work, iso-caryophyllene was found cytotoxic against in vitro culture cell lines but its mechanism of action is still unknown. Reactive oxygen species (ROS) and lipid oxidation induced by iso-caryophyllene were assessed using DCFH-DA and BODIPY-C11, respectively. The results show that iso-caryophyllene induces significant overproduction of ROS by about 187 % at 100 μM and lipid oxidation, which are both partially inhibited by - tocopherol. The effect of isocaryophyllene on membrane permeabilization was evaluated using calcein-AM assay that show that iso-caryophyllene causes membrane permeabilization and cell shrinking. -Tocopherol significantly prevents membrane permeabilization, cell shrinking and cell death, suggesting that lipid oxidation is in part implied in the cytotoxicity. Electrochemical experiments indicate that the superoxide anion reacts with iso-caryophyllene and possibly form oxidized derivatives which could initiate lipid oxidation. Interestingly, superoxide anion reacts more readily with -tocopherol in comparison with iso-caryophyllene which could explain its protective effect on cellular membrane.

Calcium Sensitization Induced by Sodium Fluoride in Permeabilized Rat Mesenteric Arteries

It was hypothesized that NaF induces calcium sensitization in Ca2+-controlled solution in permeabilized rat mesenteric arteries. Rat mesenteric arteries were permeabilized with beta-escin and subjected to tension measurement. NaF potentiated the concentration-response curves to Ca2+ (decreased EC50 and increased E(max)). Cumulative addition of NaF (4.0, 8.0 and 16 mM) also increased vascular tension in Ca2+-controlled solution at pCa 7.0 or pCa 6.5, but not at pCa 8.0. NaF-induced vasocontraction and GTPgammaS-induced vasocontraction were not additive. NaF-induced vasocontraction at pCa 7.0 was inhibited by pretreatment with Rho kinase inhibitors H1152 or Y27632 but not with a MLCK inhibitor ML-7 or a PKC inhibitor Ro31-8220. NaF induces calcium sensitization in a Ca2+-dependent manner in beta-escin-permeabilized rat mesenteric arteries. These results suggest that NaF is an activator of the Rho kinase signaling pathway during vascular contraction.

Toluene permeabilization differentially affects F- and P-type ATPase activities present in the plasma membrane of Streptococcus mutans

Streptococcus mutans membrane-bound P- and F-type ATPases are responsible for H+ extrusion from the cytoplasm thus keeping intracellular pH appropriate for cell metabolism. Toluene-permeabilized bacterial cells have long been used to study total membrane-bound ATPase activity, and to compare the properties of ATPase in situ with those in membrane-rich fractions. The aim of the present research was to determine if toluene permeabilization can significantly modify the activity of membrane-bound ATPase of both F-type and P-type. ATPase activity was assayed discontinuously by measuring phosphate release from ATP as substrate. Treatment of S. mutans membrane fractions with toluene reduced total ATPase activity by approximately 80 percent and did not allow differentiation between F- and P-type ATPase activities by use of the standard inhibitors vanadate (3 µM) and oligomycin (4 µg/mL). Transmission electron microscopy shows that, after S. mutans cells permeabilization with toluene, bacterial cell wall and plasma membrane are severely injured, causing cytoplasmic leakage. As a consequence, loss of cell viability and disruption of H+ extrusion were observed. These data suggest that treatment of S. mutans with toluene is an efficient method for cell disruption, but care should be taken in the interpretation of ATPase activity when toluene-permeabilized cells are used, because results may not reflect the real P- and F-type ATPase activities present in intact cell membranes. The mild conditions used for the preparation of membrane fractions may be more suitable to study specific ATPase activity in the presence of biological agents, since this method preserves ATPase selectivity for standard inhibitors.

Utilização de saponina em citometria de fluxo: uma alternativa factível para permeabilização celular / Use of saponin in flow cytometry: a feasible alternative to cellular permeabilization

A pesquisa de antígenos intracelulares por citometria de fluxo é essencial para o estudo imunofenotípico das doenças onco-hematológicas. A utilização de saponina para permeabilização celular tem se mostrado eficaz e de menor custo. Com suas propriedades detergentes, permeabiliza a membrana citoplasmática sem danificá-la ou alterar a expressão dos antígenos de membrana, permitindo a detecção simultânea de antígenos intracelulares e de superfície. O objetivo deste trabalho foi analisar a eficácia da técnica de permeabilização celular com a utilização de saponina como agente permeabilizante. Foram avaliadas 36 amostras de sangue periférico e de medula óssea enviadas ao Laboratório de Imunopatologia do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo para estudo imunofenotípico. Realizamos a técnica de imunofluorescência direta e utilizamos o programa CellQuest (Becton Dickinson, San Jose, CA) do citômetro de fluxo FACSCalibur (Becton Dickinson, San Jose, CA) para aquisição e análise dos dados. Observamos excesso de perda celular quando utilizamos o tampão de lavagem estocado por período superior a sete dias, à temperatura de 2ºC a 8ºC, e perda mínima com tampão de lavagem recém-preparado ou congelado no momento do preparo e descongelado apenas no momento do uso. Para eliminar o excesso de perda celular realizamos análise sistemática das etapas da técnica, o que nos permitiu adequá-la e utilizá-la na rotina de nosso laboratório. Em país de recursos escassos padronizamos uma técnica viável para pesquisar antígenos intracelulares por citometria de fluxo, mais rápida, eficaz e com redução dos custos financeiros em 35 por cento.

Intracellular antigen investigations by means of flow cytometry are essential for immune phenotypical studies of oncohematological diseases. The use of Saponin for cellular permeabilization has proved to be cost-effective. Due to its detergent properties, Saponin permeabilizes the cytoplasmatic membrane without harming it or altering membrane antigen expressions, thus allowing simultaneous detection of intracellular and surface antigens. The objective was to analyze the efficiency of the cell permeabilization technique using Saponin as a permeabilizing agent. Thirty-six samples of peripheral blood and bone marrow underwent an immune phenotypical study at the Immune Pathology Laboratory of the Hospital das Clínicas of the University of São Paulo. Direct immunofluorescence was accomplished using the CellQuest program (Becton Dickinson, San Jose, CA) of the FACSCalibur flow cytometer (Becton Dickinson, San Jose, CA) to acquire and analyze the data. Excessive cell loss was observed when using rinsing buffers stored for more than seven days at temperatures ranging from 2ºC to 8ºC. Minimal cell loss was found when rinsing buffers were used immediately after preparation or frozen at the time of preparation and thawed just before use. In order to eliminate excessive cell loss, a systematic analysis of the technique stages was carried out, allowing us to restructure the technique to our purposes and its use in our laboratory. In a country with scarce resources, a viable, faster, more cost-effective technique (35 percent reduction in cost) aimed at investigating intracellular antigens by flow cytometry has been standardized.