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• W1965718263 abstract "Among natural animal toxins, snake venom causes many fatal consequences in humans in tropical areas and is one of the most frequent dangerous contacts with these animals. Snake venom reaching the blood circulation could lead to, among other events, decreased pulse frequency, drop of blood pressure, and lethal respiratory and circulatory failure. Among the biological activities of the venom, the cardiovascular system cramping effects were most obviously detected and registered. The Brazilian Butantan Institute in Sao Paulo is world famous for its research struggle in creating life saving medicaments and antidotes. The analysis of venom produced by different snakes revealed many active components showing a broad range of therapeutical relevance. Therefore many medicaments now have snake venom-related agents. Pyroglutamyl proline-rich oligopeptides, present in the venom of the pit viper Bothrops jararaca (Bj-PROs), were the first described naturally occurring inhibitors of the angiotensin I-converting enzyme (ACE), the main target in the anti-hypertensive therapy. Recent results suggest ACE inhibition-independent mechanisms for ACE inhibitors (1, 2). Using calcium imaging by confocal microscopy and fluorescence imaging plate, Lameu et al. (in this issue) have characterized Bj-PRO-10c-induced [Ca2+]i transients in rat brain cells as being independent from bradykinin-mediated effects and ACE inhibition. By these experiments, a yet unknown receptor that induced influx and liberation of calcium from internal stores, as well as reduction of intracellular cAMP levels, was detected. The determination of the presence and number of circulating tumor cells (CTCs) is crucial for cancer therapy and follow-up. One potential assay is their magnetic enrichment and fluorescence in situ hybridization (3). Hu et al. (in this issue) have developed a multichromatic flow cytometry assay to determine the number of CTCs in breast cancer patients. The authors used a combined staining for Epithelial-cell adhesion molecule, Cytokeratin 8, 18, 19, and CD45 (to remove the abundant leukocytes). The presence of CTCs in the peripheral blood was found to be related with the presence of primary tumor or metastases and potential relationship is under analysis with the survival time and relapse of the tumor. This flow cytometry assay was proven to have higher specificity of detection of the CTCs than the RT PCR method and has the potential to be a valuable tool for prognosis assessment among breast cancer patients. Recently, Zhao et al. (4) compared histone H2AX phosphorylation with checkpoint 2 kinase (Chk2) activation in a cell culture model during topoisomerase inhibitor induced apoptosis. While H2AX phosphorylation and Chk2 activation were maximal in S-phase cells, Chk2 activation was also much pronounced in G(2)M cells; the least affected by TPT were G(1) cells. MTX-induced H2AX phosphorylation was maximal in G(1) cells, whereas Chk2 activation was maximal in G(2)M and minimal in G(1) cells. Ozawa (5) in turn reported that phosphorylation of serine on histone H3 occurs during cell cycle at G2/M transition but is also down-regulated by DNA damage. Zhao et al's recent work focuses on histone phosphorylation for the detection of DNA damage response in G1 and S-phase (in this issue). Phosphorylation of H2AX, P53, and some other proteins were detected with phospho-specific Abs and measured by laser scanning cytometry. In relation to the cell cycle phase, the induced responses were found to be selective for S-phase cells. These data are consistent with the mechanism in which stalling of DNA replication “forks” upon collision with the primary UV-induced DNA lesions and likely formation of double-strand DNA breaks triggers DDR. The prior reports (6) on induction of cH2AX in G1 cells by UV may have erroneously identified cells initiating DNA replication following UV exposure as G1 cells because of the fact that their DNA content did not significantly differ from that of G1 cells that had not initiated DNA replication. SYTO probes are a class of DNA binding fluorochromes with a great plethora of applications, whereas for many members of the SYTO class specific binding sites and application are yet to be defined as recently detailed (6). Fu et al. (in this issue) developed a flow cytometric method for analysis of the malaria inducing Plasmodium falciparum infected red blood cells (RBCs) by using SYTO61, previously used to stain nuclei of living eukaryotic cells (7). Plasmodium cells living inside the RBCs consume the hemoglobin. Reactive oxygen species (superoxide and hydrogen peroxide) are the by-products of hemoglobin digestion and are believed to exert significant oxidative stress on the parasite. The cell permeable fluorescent nucleic-acid binding SYTO 61 combined with the reduced form of fluorescent reactive oxygen species reporter 5-(and-6)-chloromethyl-2′,7′-dichlorodihy-dro-fluorescein diacetate acetyl ester was used to distinguish between uninfected and infected erythrocytes in a flow cytometric format and simultaneously characterize the amount of reactive oxygen species in RBCs. Ullal et al. (in this issue) developed a SYTO 13-based assay for microparticle (MP) detection. During the dying process, apoptotic cells compose and release membrane bound MPs. The presence of MPs on the cell's surface is a hallmark of apoptotic cells. Size of the released MPs is nearly 1 μm or smaller, therefore they do not sediment with cells and remain in the supernatant. Thereby, the supernatant could give information about the amount of MPs in a sample. Usually the side scatter parameter is used to detect MPs after apoptosis induction in the tissue culture. But in these very small size ranges, the SSC is not sensitive enough to resolve and discriminate from other small cellular particles and debris. The SYTO 13 staining of the DNA and RNA content of the MPs allowed the detection of 1.5–2.9 times as many particles as did light scatter. This increased sensitivity was observed with three different cell lines and was independent of inducing stimulus. Treatment of fixed and permeabilized MPs with DNase and RNase showed that SYTO 13 binding resulted from interaction with both DNA and RNA. Together, these findings indicate that the nucleic acid content of MPs provides the basis for their detection in in vitro systems and suggests the utility of fluorescent dyes like SYTO 13 for more sensitive quantitative assays. Atherosclerosis is frequently observed in the developed countries. Macrophages are involved in the atherosclerotic lesion development. Grandl and Schmitz (in this issue) used fluorescent high-content imaging to quantify lipid storage compartments in macrophages also in response to drugs affecting cellular lipid metabolism. E-LDL-loaded macrophages showed increased intensity of Bodipy 493/503 and LipidTOXTM-Green neutral lipid droplet staining and a greater mean area and number of lipid droplets per cell compared with Ox-LDL-loaded and M-CSF-differentiated control macrophages. In contrast, Ox-LDL-loaded macrophages show increased intensity of NBD-PE and LipidTOXTM-Red detectable phospholipidosis in the endolysosomal compartment compared with E-LDL-loaded and M-CSF-differentiated macrophages. Dr. Jozsef Bocsi, Heart Center Leipzig, is acknowledged for his help with this editorial." @default.
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• W1965718263 date "2010-03-01" @default.
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• W1965718263 title "Medicaments: Gifts from the jungle" @default.
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• W1965718263 doi "https://doi.org/10.1002/cyto.a.20874" @default.
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