Immune-regulatory properties carried by human amnion epithelial cells: Focus on the role of HLA-G and adenosinergic ectoenzymes.

Human amnion epithelial cells (hAEC) can be efficiently isolated from full-term amnion membrane and have been gaining recognition as advanced medical products. Such cells originate directly from the embryo during the early phase of development and exert a crucial function in the establishment of a tolerogenic environment, to avoid maternal immune rejection. Amnion cell immuno-modulation may be exploited, but additional efforts are required to establish the mechanisms underlying such capacity. The way to fully clarify such an issue is so far long. Here we overview current knowledge on the effects on innate or adaptive immune cells offered by intact hAEC or secreted mediators, pinpointing the mechanisms to date elucidated by our group and others. We move from the description of hAEC general features to molecular intermediaries generating effects directly or indirectly on immune cells. We focus on the role of non-canonical HLA class I molecules, with emphasis on HLA-G, but expand such analysis on adenosinergic mediators, cytokines, and hAEC-derived microvesicles. Finally, we report the ongoing clinical trials exploiting hAEC multipotency and immune modulation.

Changes in subcellular localization of visfatin in human colorectal HCT-116 carcinoma cell line after cytochalasin B treatment.

The aim of the study was to assess the expression and subcellular localization of visfatin in HCT-116 colorectal carcinoma cells after cytokinesis failure using Cytochalasin B (CytB) and the mechanism of apoptosis of cells after CytB. We observed translocation of visfatin's antigen in cytB treated colorectal carcinoma HCT-116 cells from cytosol to nucleus. Statistical and morphometric analysis revealed significantly higher area-related numerical density visfatin-bound nano-golds in the nuclei of cytB-treated HCT-116 cells compared to cytosol. Reverse relation to visfatin subcellular localization was observed in un-treated HCT-116 cells. The total amount of visfatin protein and visfatin mRNA level in HCT-116 cells was also decreased after CytB treatment. Additionally, CytB significantly decreased cell survival, increased levels of G2/M fractions, induced bi-nuclei formation as well as increased reactive oxygen species (ROS) level in HCT-116 cells. CytB treatment showed cytotoxic effect that stem from oxidative stress and is connected with the changes in the cytoplasmic/nuclear amount of visfatin in HCT-116 cells.

Exogenous administration of visfatin affects cytokine secretion and increases oxidative stress in human malignant melanoma Me45 cells.

Visfatin has recently been established as a novel adipokine that is predominantly expressed in visceral fat. Recombinant visfatin has immunomodulating properties, which can activate human leukocytes in vitro to induce cytokine production (IL-1ß, TNF-α, and IL-6). Only few studies have investigated the effect of visfatin on prostate, breast, ovarian cancer as well as astrocytoma cell biology. There have been no studies on the cytokine secretion in human melanoma cells in response to visfatin stimulation along with intracellular protein kinases inhibitors. ELISA assay was performed in supernatants of Me45 cells stimulated with visfatin in the presence or the absence of specific pharmacological inhibitors of the indicated protein kinases (p38, MEK 1, PI3k and JAK kinase) and nuclear factor kappa B (NK-κB) inhibitor. Intracellular reactive oxygen species level was measured in 2', 7'-dichlorodihydrofluorescein diacetate (H2DCF-DA)-loaded cells using a fluorescent measurement system. For determination of NF-κB activation, activated NF-κB p65 subunit was determined using an EZ-TFA-detect chemiluminescent transcription factor assay. We report that visfatin led to the significant increase in IL-6 and IL-8 level in culture supernatants of human malignant melanoma Me45 cells. Additionally visfatin resulted in the increase of the intracellular reactive oxygen species level. PI3k and NF-κB pathways were activated upon visfatin stimulation. The results may reflect the fact that PI3k pathway stimulation by visfatin may further lead to NF-κB activation and pro-inflammatory response.

Effect of acute lead intoxication on the ultrastructure of neutrophils in the peripheral blood of the rat.

The influence of lead acetate on the ultrastructure of peripheral neutrophils was investigated in adult male rats. It was found that a single intraperitoneal administration of lead at dose 150 mg Pb/kg b.w. led to ultrastructural changes in the neutrophils at 3 and 6 h post injection. At the time of testing the exposed population had a mean (+/- SD) blood lead concentration from 206.2 +/- 24.8 micrograms/100 ml to 75.2 +/- 9.9 micrograms/100 ml compared with a mean value of 4.0 +/- 0.7 micrograms/100 ml for the control groups. The ultrastructural alterations such as irregular nuclei with deep invaginations, plasma membrane pockets, the presence of vacuoles with a heterogeneous material and an increasing amount of RER cisternae, were most clearly expressed 6 h after lead administration. In addition there appeared sometimes nuclear pockets, and a prominent crystalline inclusion placed in the cytoplasmic matrix of some neutrophils. No differences in the mitochondrial morphology and cytoplasmic granule pattern between exposed and control rats were observed.

[Pulmonary microlithiasis--difference between radiology results and pulmonary function tests]. / Kamica pecherzyków plucnych--dysproporcja miedzy obrazem radiologicznym a badaniami czynnosciowymi pluc.

The case od 22-years old woman with pulmonary microlithiasis is described. The diagnosis was confirmed by histopathological examination of lung tissue. In electron microscopy lung fibrosis was seen. High resolution computed tomography (HRCT) and pulmonary function tests (lung compliance, diffusion capacity, exercise test) were done. The role of bronchoalveolar lavage (BAL) and lung scintigraphy with technetium in the diagnostic procedure of this disease is discussed.