A fast and simple method for measuring thymocyte apoptosis with propidium iodide coloring and flow chitometry.

A fast and simple method for measuring thymocyte apoptosis with propidium iodide coloring and flow chitometry.

Corticosteroids, ionoffor calcium and anti-CD3 monoclonal antibodies kill simoocyte mice incubated in vitro. Cell death is preceded by extensive DNA fragmentation into the oligonucleosomom subunit. This type of death (apoptosis), which is physiologically occurring in the intratrate process of immune cell selection, is usually evaluated by electrophoresis or colorimetric methods that measure DNA fragmentation in nuclear extracts. These techniques cannot determine the percentage of apoptotic nucleetics or recognize apoptotic cells in the population of heterogeneous cells.

We have developed a cytometric flow method to measure apoptotic nucleetic percentages after coloring propidium iodide in hypotonic buffers and has compiled it with classical columnic techniques and electrofores using Dexamethasone (DEX). ApopTotic Nuclei emerged as the culmination of extensive hypodiploid DNA which was easily discriminated against from the narrow peak of thymocytes with a normal (diploid) DNA content in the red fluorescence channel. When Dex induced apoptosis is inhibited by low temperature incubation (4 degrees C) or cycloheximide treatment, no peak of the hypodiploid DNA appears.

Likewise, thymocyte death caused by sodium azide, a substance with cell murder activities through non-apoptotic mechanisms, does not produce any variations in the peak of normal DNA. Sitometric flow data shows a very good correlation with the results obtained by electrophoretic and colorimetric methods. This new, simple and simple method must prove useful to assess certain population apoptosis in heterogeneous tissue such as bone marrow, thymus and lymph nodes.

Macrophage alternative activation: Immunological functional perspective.

Macrophages are default immune cells with established roles in the main response to pathogens, but also in network homeostasis, coordination of adaptive immune response, inflammation, resolution, and repairs. These cells recognize danger signals through receptors that are able to induce special activation programs. Activation of macrophages known to be induced classically by IFN-Gamma, which triggered a hard proinflammatory response needed to kill intracellular pathogens.

Macrophages also undergo alternative activation by IL-4 and IL-13, which trigger different phenotypes that are important for immune responses to parasites. Here we review this cytokine cellular source, receptor signal path, and mark markers and genutors of genes. We draw attention to differences found between the mouse and alternative human activation model. Evidence for in vivo alternative macrophages activation was also analyzed, with nematode infection as a prototypic disease. Finally, we revisit the concept of macrophage activation in the context of the immune response.

In the analysis of Vivo Autophagy in response to nutritional hunger using transgenic mice that expressed the automophagosome fluorescent marker.

Macroautophagy mediates mass degradation of cytoplasmic components. It accounts for degradation of the longest protein aged: cytoplasmic constituents, including organelles, exiled to autofagosomes, which are then fuses with lisosomes, where relegation occurs. Although the possibility of autophagy involvement in homeostasis, development, cell death, and pathogenesis has been repeatedly shown, systematically in Vivo analysis has not been carried out in mammals, especially due to limited monitoring methods.

To understand where and when autophagy occurs in Vivo, we have produced transgenic mice that express GFP systemically integrated with LC3, which is a mammalian homologous from ATG8 yeast (AUT7 / APG8) and serves as a marker protein for autophagosomes. Fluorescence microscopic analysis reveals that different autophagies are induced by nutrean hunger in most tissues.

A fast and simple method for measuring thymocyte apoptosis with propidium iodide coloring and flow chitometry.

On several networks, Autophagy even occurs actively without starvation. Our results show that autophagy regulations depend on organs and the role of autophagy are not limited to hunger response. This transgenic mouse model is a useful tool for studying autophagy mammals.

The generation of functional milk glands of single stem cells.

The existence of mammary stem cells (Mascs) has been postandaken from evidence that the mammary gland can be regenerated by transplantating epithelial fragments in mice. Interest in Mascs has been further distimulated with their potential role in breast tumorigenesis. However, the identity and purification of Mascs has proven difficult to understand because of the lack of specified markers.

We isolate the discrete population of mammarus cells based on cell surface markers and identify subpopulations (LIN-CD29HICD24 +) which are highly enriched for mascs with transplantation. Here we show that one cell, is marked with Transgene Lacz, can reorganize the complete mammary gland in Vivo.

Transplant cells contribute to the lines of luminal and myoepithelial and generated functional lobuloalveolar units during pregnancy. The capacity of the destruction of these cells is indicated by serial transplants from clone results. In supporting the potential role for Mascs in breast cancer, subpared subpopulation by the parent is expanded in premalignant milk tissue from MMTV-WNT-1 mice and contains a higher number of mascs.

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Our data specifies that single cells in the LIN-CD29HICD24 + population are multipotent and own updates, the properties that define them as Mascs.

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