Expression of interleukine-8 as an independent prognostic factor for sporadic colon cancer dissemination.

AIM: The aim of our study was to investigate the gene and serum protein expression profiles of IL-8 in colon cancer and associated hepatic metastasis and to correlate these results with clinicopathologic variables of the patients. MATERIALS AND METHODS: IL-8 was evaluated by qPCR and ELISA in a total number of 62 colon cancer patients (n=42 by qPCR and n=20 by ELISA) in normal and tumoral tissue specimens and serum samples respectively. Additionally synchronous metastasis from 5 of these patients were also collected at the time of surgery and analyzed by qPCR. RESULTS: IL-8 was up regulated in all analyzed tumoral samples compared with normal tissue (P-value = 0.01) and higher expressed in metastatic tissues compared with tumoral tissues (P -value= 0.03). The median expression of IL-8 in patients over 60 years old was found to be higher compared with the median expression of IL8 in patients less than 60 years old (3.89 compared with 14.69, P -value= 0.005). According to tumor grading, we found that IL-8 in tumors with well differentiated adenocarcinoma have a median mRNA expression of 9.78 compared with a median mRNA IL8 expression of 26.63 in moderate or poor differentiated adenocarcinoma. Levels of IL-8 determined in serum were statistically significant correlated with preoperative carcinoembryonic antigen level (P -value= 0.003, R=0.57) and with distant metastasis (P-value =0.008). Serum level of IL-8 increased proportionally along with TNM tumor stage and was found to be statistically significant correlated with C-reactive protein (P -value, R=0.64). Colon cancer patients had higher IL-8 levels as determined by ELISA (median value= 29.64 pg/ml) compared with healthy controls (median value= 4.86 pg/ml). DISCUSSIONS: Our results provide additional support for the role of inflammation in colon cancer and indicate that IL-8 could be further validated in association with other already used markers for prognostic and diagnostic of evolutional disease in colon cancer patients.

Stimulatory and possible antioxidant effects of High Density Green Photons (HDGP) on cellular systems.

The interactions between the electromagnetic field and the biological systems were extensively investigated, with remarkable results and advanced technologies. Nevertheless, the visible domain of the spectrum has been rather neglected, since the classic physics did not allow electronic transitions induced by visible light. Recently, the interaction of light with the matter has generated a new scientific domain known in Physics as optical manipulation, with the new concepts of optical matter and optical force. This article presents the results of our work concerning in vitro effects of High Density Green Photons (HDGP) irradiation on cell cultures: stimulation of cell proliferation and migration and a possible antioxidant action.

Prognostic and predictive potential molecular biomarkers in colon cancer.

An important objective in nowadays research is the discovery of new biomarkers that can detect colon tumours in early stages and indicate with accuracy the status of the disease. The aim of our study was to identify potential biomarkers for colon cancer onset and progression. We assessed gene expression profiles of a list of 10 candidate genes (MMP-1, MMP-3, MMP-7, DEFA 1, DEFA-5, DEFA-6, IL-8, CXCL-1, SPP-1, CTHRC-1) by quantitative real time PCR in triplets of colonic mucosa (normal, adenoma, tumoral tissue) collected from the same patient during surgery for a group of 20 patients. Additionally we performed immunohistochemistry for DEFA1-3 and SPP1. We remarked that DEFA5 and DEFA6 are key factors in adenoma formation (p<0.05). MMP7 is important in the transition from a benign to a malignant status (p <0.01) and further in metastasis being a prognostic indicator for tumor transformation and for the metastatic potential of cancer cells. IL8, irrespective of tumor stage, has a high mRNA level in adenocarcinoma (p< 0.05). The level of expression for SPP1 is correlated with tumor level. We suggest that high levels of DEFAS, DEFA6 (key elements in adenoma formation), MMP7 (marker of colon cancer onset and progression to metastasis), SPP1 (marker of progression) and IL8 could be used to diagnose an early stage colon cancer and to evaluate the prognostic of progression for colon tumors. Further, if DEFA5 and DEFA6 level of expression are low but MMP7, SPP1 and IL8 level are high we could point out that the transition from adenoma to adenocarcinoma had already occurred. Thus, DEFA5, DEFA6, MMP7, IL8 and SPP1 consist in a valuable panel of biomarkers, whose detection can be used in early detection and progressive disease and also in prognostic of colon cancer.

Cyclosporin A induces an atypical heat shock response.

Cyclosporin A is a widely used immunosuppressive drug having toxic side effects, in particular on kidneys and liver, as a result of its action on different molecular targets. Here we demonstrate that low doses of CsA are able to induce the expression of the heat shock protein HSP27 and its hyperphosphorylation. It also activates the two heat shock transcription factors, HSF1 and HSF2. Since these factors have been shown to be activated by proteasome inhibition, we tested the hypothesis that the inhibitory action of CsA on the proteasome might be responsible for the activation of HSFs and the subsequent expression of HSP27. The increase in multiubiquitinated proteins as well as the stabilization of p53 following CsA addition argues in favor of this hypothesis. The kidney BSC-1 cells are highly responsive to the addition of CsA: the possible link between HSP27 induction and hyperphosphorylation and nephrotoxicity is discussed.

MAP kinase activation by cyclosporine A.

Short treatment of HeLa cells with cyclosporine A led to the activation in the crude cell extracts of a MAP kinase-like activity. Fractionation by chromatography on a Mono Q column allowed the separation of two activities co-eluting with the MAP kinases ERK1 and ERK2. The activation of these two MAP kinases was demonstrated in Western Blotting by the appearance, after CsA treatment, of two new slowly migrating forms on SDS electrophoretic gels. A similar activation was also obtained in renal epithelial BSC-1 cells and 3T3 fibroblasts. MAP kinase activation might result from a perturbation of calcium homeostasis induced by CsA treatment.

GRP78 induction by cyclosporin A in human HeLa cells.

Immunosuppressive drugs such as cyclosporin A (CsA) and FK506 are known to have pleiotropic effects on cells. Here we demonstrate that treatment of HeLa cells with low concentrations of CsA (but not of FK506) induces the synthesis of a stress protein, GRP78, located inside the endoplasmic reticulum. High concentrations of CsA lead to a general decrease in protein synthesis. When cells are stressed (heat-shocked) during the CsA treatment, the synthesis of heat shock proteins is reinforced. FK506 has no detectable effects at any concentration. The mechanism of induction of GRP78 by CsA remains presently unknown. Whatever the mechanism involved, GRP78 overexpression might be responsible for some of the physiological effects of CsA.

The effect of diapulse therapy on the healing of decubitus ulcer.

The effect of pulsed high peak power electromagnetic field (Diapulse) on treatment of pressure ulcers is under investigation. 20 elderly patients, aged from 60 to 84, hospitalized with chronic conditions and bearing long-standing pressure ulcers, are subjected to Diapulse sessions (1-2 daily), parallel to conventional treatment. 5 patients undergo conventional therapy, serving as control and 5 others follow conventional+placebo Diapulse treatment. All patients were daily monitored, concerning their clinical status and ulcers' healing. After a maximum 2-weeks treatment, bulge healing rate was, as follows: 85% excellent and 15% very good healing under Diapulse therapy; in the placebo group, 80% patients show no improvement and 20% poor improvement; in the control group, 60% patients show no improvement and 40% poor improvement of ulcers. This investigation strongly advises for Diapulse treatment as a modern, uninvasive therapy of great efficiency and low social costs in resolving a serious, widespread medical problem.

The effect of pulsed electromagnetic field (Diapulse) on cellular systems.

The effect of a 27.12 MHz pulsed electromagnetic field (Diapulse) on microbial growth is investigated. A strain of K 12 E. coli grown in complete Pennassay medium is subjected to Diapulse action for 30 min, at 8 hrs and 12 hrs of growth. In this experimental set-up, designed to be closed to the physiological conditions of open wounds, the Diapulse action does not promote any increase of cell population, indicating the safety of this type of therapy for wound healing process. The same K 12 E. coli strain grown in Pennassay medium for 2 hours is inoculated into a minimal growth medium and the lagless exponential growth thus obtained is followed by a spectrophotometric method. Diapulse field is applied to this lagless phase of cellular cultures at 30, 60, and 90 minutes after inoculation. A slight increase in the number of cells was observed at 2 and 4 hours after the Diapulse application, when the cultures were previously subjected to Diapulse action between the period of 60 and 90 minutes of their growth. A possible molecular mechanism for these effects is discussed.

The stimulation of fibronectin synthesis by high peak power electromagnetic energy (Diapulse).

The effect of Diapulse therapy (pulsed electromagnetic energy of 27.12 MHz frequency) on post-operative wound healing and plasma fibronectin concentration is investigated. Patients treated with Diapulse, locally and over hepatic area present higher fibronectin levels starting 3d day after surgery. These higher values correlate well with a clear improvement of wound healing processes.

The influence of pulsed high peak power electromagnetic energy (Diapulse) treatment on posttraumatic algoneurodystrophies.

Forty-five patients with post-traumatic algoneurodystrophies were treated with Diapulse therapy (high peak power electromagnetic energy). This therapy accelerates the healing of the disease inducing a recovery of the damaged bone structure.

The effects of pulsed, high frequency radio waves on rat liver (ultrastructural and biomedical observations).

The effects of a high frequency electro-magnetic field, generated by a Diapulse instrument (Diapulse Corporation of America) on rat liver has been investigated. Ultrastructural aspects are described and quantitative determinations of mitochondrial enzymes MAO, CyT-Ox, MDH, SDH and ATP-ase recorded. The standard therapeutic parameters generally used with the Diapulse instrument in medicine were found to induce a stimulation effect at the investigated level, without apparent degenerative modifications. A concordance between the qualitative ultrastructural data and quantitative subcellular enzymic determinations has been observed.