Extracellular ATP induces cytokine expression and apoptosis through P2X7 receptor in murine mast cells

Extracellular ATP and other nucleotides act through specific cell surface receptors and regulate a wide variety of cellular responses in many cell types and tissues. In this study, we demonstrate that murine mast cells express several P2Y and P2X receptor subtypes including P2X(7), and describe functional responses of these cells to extracellular ATP. Stimulation of bone marrow-derived mast cells (BMMC), as well as MC/9 and P815 mast cell lines with millimolar concentrations of ATP, resulted in Ca(2+) influx across the cellular membrane and cell permeabilization. Moreover, brief exposures to ATP were sufficient to induce apoptosis in BMMCs, MC/9, and P815 cells which involved activation of caspase-3 and -8. However, in the time period between commitment to apoptosis and actual cell death, ATP triggered rapid but transient phosphorylation of multiple signaling molecules in BMMCs and MC/9 cells, including ERK, Jak2, and STAT6. In addition, ATP stimulation enhanced the expression of several proinflammatory cytokines, such as IL-4, IL-6, IL-13, and TNF-alpha. The effects of ATP were mimicked by submillimolar concentrations of 3-O-(4'-benzoyl)-benzoyl-benzoyl-ATP, and were inhibited by pretreatment of mast cells with a selective blocker of human and mouse P2X(7) receptor, 1[N,O-bis(5-isoquinolinesulphonyl)-N-methyl-l-tyrosyl]-4-phenylpiperazine, as well as oxidized ATP. The nucleotide selectivity and pharmacological profile data support the role for P2X(7) receptor as the mediator of the ATP-induced responses. Given the importance of mast cells in diverse pathological conditions, the ability of extracellular ATP to induce the P2X(7)-mediated apoptosis in these cells may facilitate the development of new strategies to modulate mast cell activities.     

How a cytokine is chaperoned through the secretory pathway by complexing with its own receptor: lessons from interleukin-15 (IL-15)/IL-15 receptor alpha

While it is well appreciated that receptors for secreted cytokines transmit ligand-induced signals, little is known about additional roles for cytokine receptor components in the control of ligand transport and secretion. Here, we show that interleukin-15 (IL-15) translocation into the endoplasmic reticulum occurs independently of the presence of IL-15 receptor α (IL-15Rα). Subsequently, however, IL-15 is transported through the Golgi apparatus only in association with IL-15Rα and then is secreted. This intracellular IL-15/IL-15Rα complex already is formed in the endoplasmic reticulum and, thus, enables the further trafficking of complexed IL-15 through the secretory pathway. Just transfecting IL-15Rα in cells, which transcribe but normally do not secrete IL-15, suffices to induce IL-15 secretion. Thus, we provide the first evidence of how a cytokine is chaperoned through the secretory pathway by complexing with its own high-affinity receptor and show that IL-15/IL-15Rα offers an excellent model system for the further exploration of this novel mechanism for the control of cytokine secretion.     

Effect of a pseudovolume image in multifocus radiography

The effect of a pseudovolume image was theoretically substantiated on the basis of calculations. This was experimentally studied using with microfocus x-ray diagnostic apparatuses. Digital microfocus and standard x-ray study were compared using an experimental material. Microfocus x-ray study is a highly informative technique that shows a greater possibility to provide volume images of an object than does standard x-ray study.     

Capabilities of digital microfocal x-ray study in the evaluation of reparative regeneration of bone tissue in an experiment

Digital microfocal x-ray study was experimentally studied in animals to examine the time course of changes in their bone regeneration. Sixteen Chinchila rabbits whose bone defect in the angle of the mandibular ramus had been closed with the osteoplastic material Gapcol with the applied allogeneic, autologous stem cells isolated from rabbit adipose tissue and human plasma enriched with thrombocytic growth factors were examined. The capabilities of digital microfocal x-ray study versus x-ray computed tomography were compared in the evaluation of reparative regeneration of bone tissue. The results of radiation studies were verified with the data of scanning electron microscopy.     

Novel aryl and heteroaryl substituted N-[3-(4-phenylpiperazin-1-yl)propyl]-1,2,4-oxadiazole-5-carboxamides as selective GSK-3 inhibitors

Synthesis, biological evaluation, and SAR dependencies for a series of novel aryl and heteroaryl substituted N-[3-(4-phenylpiperazin-1-yl)propyl]-1,2,4-oxadiazole-5-carboxamide inhibitors of GSK-3beta kinase are described. The inhibitory activity of the synthesized compounds is highly dependent on the character of substituents in the phenyl ring and the nature of terminal heterocyclic fragment of the core molecular scaffold. The most potent compounds from this series contain 3,4-di-methyl or 2-methoxy substituents within the phenyl ring and 3-pyridine fragment connected to the 1,2,4-oxadiazole heterocycle. These compounds selectively inhibit GSK-3beta kinase with IC(50) value of 0.35 and 0.41 microM, respectively.     

Spiral computed tomography in the diagnosis of limb osteomyelitis

The results of radiation studies in 121 patients of different age (4 to 75 years) examined for limb osteomyelitis are analyzed. All the patients underwent routine X-ray study and computed tomography (CT), 26 patients had X-ray fistulography; 8, linear tomography; 10, CT fistulography; 6, scintigraphy, and 15, ultrasound study. Acute hematogenous osteomyelitis (AHO), chronic hematogenous osteomyelitis (CHO), and atypical (here Garre's sclerosing osteomyelitis and Brodie's abscess) osteomyelitis were ascertained in 10.6, 26.4, and 10.1% of cases, respectively. Posttraumatic osteomyelitis was diagnosed in almost 50% of the patients. CT defined the phase of chronic limb osteomyelitis. Spiral CT has proven to be the most effective technique for diagnosing limb osteomyelitis as compared with routine X-ray study: the accuracy of X-ray study was 81.8%, its sensitivity, 84.9%, and specificity, 60.0% and those of computed tomography were 96.7, 99.1, and 80.0%, respectively.     

Standardizing a protocol of magnetic resonance imaging of temporomandibular joints. Part 2. Unification of analysis of obtained data

The paper presents a unified protocol for analyzing the data obtained by magnetic resonance tomography, which has been used to examine 350 patients. It characterizes the MR semiotics of different pathological conditions of articular structures, which are illustrated by MR images. An optimal terminology is proposed for the evaluation of bone and soft tissue changes.