High Doses of Ascorbate Kill Y79 Retinoblastoma Cells In vitro

Objectives: To tests the sensitivity of Y79 retinoblastoma cell lines to high doses of ascorbate, in vitro, and compare its effects with those of some chemotherapeutic agents routinely employed in the treatment of retinoblastoma. Methods: Y79 retinoblastoma cells have been exposed to increasing doses of either sodium ascorbate (SA) or Melphalan (MEL), to define a dose-response curve around the peak plasma concentrations reached by both chemicals when administered according to the existing therapeutic procedures and protocols. The assessment of cell number and viability was performed, before and after exposure, with both the manual (Trypan Blue Exclusion Test) and automated (flow cytometry) methods. Fluorescence microscopy and direct observation of cells in culture, with inverted microscope, were also performed. Results: Y79 cells are highly sensitive to the cytotoxic effect of SA, with cell viability reduced of over 90% in some experiments. As reported in the literature, this effect is directly cytotoxic and most probably mediated by acute oxidative stress on different cellular components. The same does not apply to Melphalan which, at the doses commonly used for therapeutic purposes, did not show any significant effect on cell viability, in vitro. Conclusion: To our knowledge, this is the first report showing that high doses of SA can actively kill retinoblastoma cells in vitro. While it is not surprising for SA, to show direct cytotoxic effect on tumor cells, the data reported herein represent the first evidence in favor of the possible clinical use of high doses of intravenous SA, to treat children affected by retinoblastoma. Given the many advantages of SA over the chemotherapeutic agents commonly employed to treat cancer (including its almost total absence of toxic or side effects, and its exclusive specificity for cancer cells), it is reasonable to assume, from the data reported herein, that the high doses of intravenous ascorbate, have the potential to represent a real revolution in the treatment of retinoblastoma

Muscle pathology patterns in possibly adjuvant related autoimmune/inflammatory syndrome (ASIA)

Growing evidence shows a link for biologically inert molecules, such as vaccine adjuvants and silicone implants, with the occurrence of autoimmunity-related disorders, defined as autoimmune/inflammatory syndrome induced by adjuvant-ASIA (1). Clinical conditions encompass siliconosis, the Gulf war syndrome, the macrophagic myofasciitis syndrome (MMF), post-vaccination phenomena and the spectrum of related syndromes is expanding (2). Involvement of skeletal muscle in ASIA is acknowledged in MMF, defined by long-term persistence of vaccine alum adjuvants within macrophages at sites of previous immunization. A few reports describe vaccine and silicone implants related autoimmune inflammatory myopathies (3). We carried out an immunopathological analysis of skeletal muscle biopsy in a case of MMF and two cases of possible ASIA myositis, chronologically subsequent to breast silicone implant. MMF showed the typical fascial/ perimysial macrophagic invasion, with no endomysial mononuclear infiltrates and fibral neolocalization of MHC-I complex restricted to the adjacency of macrophage deposits. The first myositis case presented with a subacute onset twenty years after an uneventful additive breast silicone implant. Endomysial inflammation, microangiopathy and multifocal fibral localization of MHC-II complex were observed. In the second patient, the onset of proximal weakness, myalgiae and a tenfold increase of creatinkinase levels occurred seven years after an unsuccessful additive mastoplasty, with rupture of prostheses and re-implantation three years later. Muscle biopsy, besides inflammation changes, showed peculiar myofibrillar disruption, with MHC-I reactive sarcoplasmic inclusions expressing several structural muscle proteins. Molecular pathogenesis of ASIA is yet undefined: genetical susceptibility is currently investigated (1,2). Due to the role of vaccines in medicine and the wide use of silicon medical devices, identification of their cause/effect link with autoimmunity is of great interest

The physiological interferon response. V. Antiviral activity present in rat lymph is neutralized by anti-mouse interferon-gamma antibodies

By neutralization tests using anti-rat and mouse interferon (IFN) antibodies, we tested whether the antiviral activity present in abdominal lymph but absent in plasma of healthy rats, could be ascribed to IFN. Antimouse IFN-gamma antibodies neutralized the inhibitor completely while anti-rat IFN-alpha/beta antibodies did not. We conclude that rat lymph contains traces of IFN-gamma and that the antiviral activity is not due to low-density lipoproteins, immunoglobulins, or to cell-produced viral inhibitors. This finding extends previous observations on rabbit lymph and further supports the existence of a physiological low-level interferon response

Athletic humans and horses: Comparative analysis of interleukin-6 (IL-6) and IL-6 receptor (IL-6R) expression in peripheral blood mononuclear cells in trained and untrained subjects at rest

Background Horses and humans share a natural proclivity for athletic performance. In this respect, horses can be considered a reference species in studies designed to optimize physical training and disease prevention. In both species, interleukin-6 (IL-6) plays a major role in regulating the inflammatory process induced during exercise as part of an integrated metabolic regulatory network. The aim of this study was to compare IL-6 and IL-6 receptor (IL-6R) mRNA expression in peripheral blood mononuclear cells (PBMCs) in trained and untrained humans and horses.Results Nine highly trained male swimmers (training volume: 21.6 ± 1.7 h/wk in 10-12 sessions) were compared with two age-matched control groups represented by eight lightly trained runners (training volume: 6.4 ± 2.6 h/wk in 3-5 sessions) and nine untrained subjects. In addition, eight trained horses (training volume: 8.0 ± 2.1 h/wk in 3-4 sessions) were compared with eight age-matched sedentary mares. In humans, IL-6 mRNA levels in PBMCs determined by quantitative reverse transcription-polymerase chain reaction were significantly higher in highly trained subjects, whereas IL-6R expression did not differ among groups. In horses, transcripts of both IL-6 and IL-6R were significantly up-regulated in the trained group.Conclusions Up-regulation of IL-6R expression in PBMCs in horses could reflect a mechanism that maintains an adequate anti-inflammatory environment at rest through ubiquitous production of anti-inflammatory cytokines throughout the body. These findings suggest that the system that controls the inflammatory response in horses is better adapted to respond to exercise than that in humans

Endothelin receptor A expression in human inflammatory cells.

Abstract Most inflammatory diseases show elevated levels of endothelin-1 (ET-1) probably due to an alteration in vascular structure and function with activation/accumulation of inflammatory cells. The ET receptors (ETA, ETB) are widely expressed in all human vessels, consistent with the main role of ET-1 in maintaining physiological vascular tone. Previous findings have shown the expression on inflammatory cells such as neutrophils (PMNs) and macrophages (MØs) of ET-1 and endothelin-converting enzyme-1 (ECE-1) (the key enzyme in the biosynthesis of ET-1). Therefore the role of ET-1 cannot be related only to the vasoactivity. Our study was aimed to determine the expression and the cellular location of ET receptors in both human PMNs and MØs by the use of RT-PCR assay, Western blot analysis and immunocytological methods. Our results showed for the first time that PMNs and MØs clearly expressed ETA (mRNA and protein). Considering that the overproduction of ET-1 following endothelial dysfunction and inflammation, contributes to pathophysiological processes such as vascular hypertrophy, cell proliferation and fibrosis, our results suggest that PMNs and MØs can also play a key role in vascular dysfunctions via the possible formation of an autocrine loop between ET-1 and ETA

Degradation of human 125I-interferon alpha by isolated perfused rabbit kidney and liver.

Cells attached to intrauterine devices (IUDs) release interferon (IFN) in the medium during incubation in vitro. Most of the IFN is released during the first hour suggesting that the cells had been previously induced and were probably already producing IFN in vivo. Characterization of the IFN indicates that most of it is of gamma-type with a trace of alpha. Production of IFN in the uterine fluid would represent a first example of the physiological IFN response and may serve to modulate some of the mechanisms preventing implantation of blastocyst. The actual presence of IFN in the uterine secretion remains to be demonstrated. PIP: IUDs were removed under aseptic conditions from 16 women at the 5th-6th day of the menstrual cycle after having been in situ for periods of 20-26 months. After removal, the tails of the devices were eliminated and cells attached to the IUDs were treated and analyzed. About 78% of the cells were macrophages, 19% neutrophils, 3% lymphocytes. The highest amount of interferon (IFN) was received from the medium during the 1st 80 minutes of incubation. There is a maximum increase in the kinetic of IFN release during the 1st hour followed by a slow progressive increase for the next 24 hours when it tends to level off. Antiviral activity was considerably acid-labile, insensitive to antihuman IFN-beta serum, slighly sensitive to antihuman IFN-alpha serum, and markedly sensitive to antihuman IFN-gamma serum. This indicates that cells attached to IUDs can produce IFN-gamma and probably a little IFN-alpha. The antiviral activity of IFN may also be useful in preventing spread of opportunistic viruses, and fungus infections in the endometrium. The presence of IFN in the uterine fluid may help modulate several mechanisms preventing implantations of the blastocyst