Mitochondrial production of oxidants contributes to a variety pathological conditions including the vascular complications diabetes, neurodegenerative diseases, and cellular senescence. We postulated that transcriptional coactivator, peroxisome proliferator activated receptor-gamma coactivator 1alpha (PGC-1alpha), major regulator oxidative metabolism mitochondrial biogenesis, could be involved in regulation antioxidant defense system endothelial cells.We show PGC-1alpha is present human,...

Incubation of ex vivo cultured mature B cells in the presence nitric oxide or oxide-donor substances delays programmed cell death as determined by appearance DNA laddering agarose gel electrophoresis flow-cytometry analysis DNA. Nitric also rescues from antigen-induced apoptosis but fails to provide a co-stimulatory signal that converts elicited antigen into proliferative response. The protective effects against can be reproduced treatment with permeant analogues cyclic GMP. Regarding...

Treatment of fetal rat hepatocytes with transforming growth factor beta (TGF-beta) is followed by apoptotic cell death. Analysis radical oxygen species (ROS) content and mitochondrial transmembrane potential (Deltapsim), using specific fluorescent probes in FACScan confocal microscopy, showed that TGF-beta mediates ROS production precedes the loss Deltapsim, release cytochrome c, activation caspase 3. induces a decrease protein mRNA levels bcl-xL, an antiapoptotic member Bcl-2 family. In...

Transforming growth factor-β (TGF-β), a regulator of fetal hepatocytes in primary culture, also regulates death these cells. Dose-response analysis showed that the TGF-β concentration needed to induce hepatocyte (2.5 ng/ml) was 5 times inhibit cells (0.5 ng/ml). In response TGF-β, induced DNA fragmentation and appearance nuclei with content lower than 2C (diploid content), typical programmed cell model. TGF-β-induced apoptosis preceded by an induction reactive oxygen species production...

p38 alpha mitogen-activated protein (MAP) kinase is a broadly expressed signaling molecule that participates in the regulation of cellular responses to stress as well control proliferation and survival many cell types. We have used lines derived from knockout mice study role this pathway apoptosis. Here, we show cardiomyocytes fibroblasts lacking are more resistant apoptosis induced by different stimuli. The reduced alpha-deficient cells correlates with decreased expression mitochondrial...

AP-1 (Activating Protein 1) transcription factor activity is tightly regulated at multiple levels, including dimer formation (i.e., Fos/Jun). Here we show that the intermediate filament protein lamin A/C suppresses function through direct interaction with c-Fos, and both proteins can interact colocalize nuclear envelope (NE) in mammalian cells. Perinuclear localization of c-Fos absent Lmna -null cells but be restored by A overexpression. In vitro, preincubation prior to addition c-Jun...

Abstract Resveratrol (RES), a chemopreventive molecule, inhibits the proliferation of tumor cells different etiologies. We previously showed that RES alters cell cycle and induces apoptosis in MCF‐7 breast by interfering with estrogen receptor (ERaα)–dependent phosphoinositide 3‐kinase (PI3K) pathway. Here, we analyzed signaling downstream PI3K, to understand mechanisms RES‐induced apoptosis. Apoptotic death was mediated Bcl‐2 downregulation since overexpression this protein abolished...

A group of 22 unrelated patients with sporadic or non-X-linked Alport syndrome were screened for mutations in the non-collagenous domain type IV collagen alpha 3 (COL4A3) chain gene. The five 3'-exons this gene, located on chromosome 2qter, tested by single strand conformation polymorphism analysis and direct sequencing. One patient was heterozygous another homozygous (Mochizuki et al., Nature Genetics, press) a deletion nucleotides. third appeared to be compound heterozygote two different...

Resveratrol is a polyphenol found at high concentrations in grapes and red wine with reported anticarcinogenic effects. We studied the molecular mechanism of resveratrol-induced apoptosis proliferation arrest prostate derived cells PZ-HPV-7 (nontumorigenic line), LNCaP (androgen-sensitive cancer PC-3 (androgen-insensitive line). Apoptosis cell cycle distribution were evaluated by flow cytometry MTT assay direct counting. Caspases, bax, bcl-2, cyclins, Cdks, p53, p21, p27 measured Western...

The multikinase inhibitor sorafenib is the only effective drug in advanced cases of hepatocellular carcinoma (HCC). However, response differs among patients and effectiveness implies a delay. We have recently described that sensitizes HCC cells to apoptosis. In this work, we explored six different liver tumor cell lines define phenotypic signature may predict lack patients. Results indicated show mesenchymal-like phenotype, resistance suppressor effects transforming growth factor beta...

Abstract Seminal extracellular vesicles (EVs) include exosomes (ø 40–120 nm) and microvesicles (MVs, ø 120–1000 nm), which would be involved in multiple functional reproductive roles. The study aimed to establish EV subtypes are present pig semen, using a high-resolution flow cytometer explore differences their tetraspanin expression profile. EVs were isolated from 12 ejaculates serial ultracentrifugation characterized by dynamic light scattering electron microscopy for size morphology as...

Abstract Background Porcine seminal plasma (SP) is endowed with a heterogeneous population of extracellular vesicles (sEVs). This study evaluated the immunophenotypic profile by high-sensitivity flow cytometry eight sEV subpopulations isolated according to their size (small [S-sEVs] and large [L-sEVs]) from four different SP sources, namely three ejaculate fractions (the first 10 mL sperm rich fraction [SRF-P1], remaining SRF [SRF-P2], post-SRF [PSRF]) entire (EE). Methods Seminal EVs were...

Treatment of elicited peritoneal macrophages or the macrophage cell line RAW 264.7 with high concentrations nitric oxide donors is followed by apoptotic death. Analysis changes in mitochondrial transmembrane potential (ΔΨm) specific fluorescent probes showed a rapid and persistent increase ΔΨm, that usually decreases cells undergoing apoptosis through mitochondrial-dependent mechanisms. Using confocal microscopy, release cytochrome c from mitochondria to cytosol was characterized as an early...

Store-operated calcium entry (SOCE) is an important Ca2+ pathway that regulates many cell functions. Upon store depletion, STIM1, a transmembrane protein located in the endoplasmic reticulum (ER), aggregates and relocates close to plasma membrane (PM) where it activates store-operated channels (SOCs). Although STIM1 was early defined as phosphoprotein, contribution of phosphorylation has been elusive. In present work, found be target extracellular-signal-regulated kinases 1 2 (ERK1/2) vitro,...

Abstract Modulation of macrophage survival is a critical factor in the resolution inflammatory responses. Exposure to LPS protects innate immune cells against apoptosis, although precise pathways responsible for prolongation remain be fully established. The goal this study was characterize mechanism TLR4-mediated murine bone marrow-derived macrophages upon M-CSF withdrawal more detail. Using combination knockout mice and pharmacological inhibitors allowed us show that TLR4 TLR2 stimulation...

Seminal plasma contains many morphologically heterogeneous extracellular vesicles (sEVs). These are sequentially released by cells of the testis, epididymis, and accessory sex glands involved in male female reproductive processes. This study aimed to define depth sEV subsets isolated ultrafiltration size exclusion chromatography, decode their proteomic profiles using liquid chromatography-tandem mass spectrometry, quantify identified proteins sequential window acquisition all theoretical...

Abstract Activation of the macrophage cell line RAW 264.7 with LPS and IFN-γ induces apoptosis through synthesis high concentrations NO due to expression synthase-2. In addition NO, activated macrophages release other molecules involved in inflammatory response, such as reactive oxygen intermediates PGs. Treatment cyclopentenone PGs, which are synthesized late onset, exerted a negative regulation on activation by impairing genes host defense, among them However, despite attenuation...