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Cancer progression is associated with significant cachexia-induced weight loss and stomatitis. Pentoxifylline (PTX) is a drug shown to have beneficial anti-inflammatory effects in cancer patients, mainly through anti-TNFα mechanisms. This study determined the PTX effects and mode of action on weight-loss, stomatitis, and survival in colon cancer patients treated with chemotherapy, examining the kinetics of tumor markers and cytokine levels.Forty patients with metastatic colon cancer receiving chemotherapy, were randomized in this study. Seventeen patients were assigned to the treatment group - 8 received a full PTX dose (400 mg TID) and 9 a reduced dose (200 mg TID). Results were compared to 23 untreated, control patients. Blood analysis of tumor markers (CEA and TPS), inflammatory cytokines (IL-1β, IL-6, IL-8, TNFα, TNF-R), CRP and sIL-2R, were performed. Additionally, clinical parameters were assessed.Patients treated with PTX (full/reduced doses), gained significant weight, and experienced a reduction in stomatitis, resulting in multiple beneficial effects, including improved life quality. Significant reductions in CRP, sIL-2R, and inflammatory cytokine levels, correlated to increases in weight and a reduction in stomatitis. A similar pattern was observed in tumor marker levels, where decreasing levels were correlated with weight gain and reduction in inflammatory cytokine levels.Colon cancer patients receiving PTX with chemotherapy, experienced weight gain and reduced stomatitis occurrence. Beneficial PTX effects were correlated to significant decreases in patient inflammatory cytokines and tumor marker levels, probably due to PTX mode of action.

BackgroundIrradiation, which affects cytokine secretion, is used to treat cancer patients. Cytokine levels have correlations to disease parameters, serving as biomarkers for patients. We aim to explore the effect of irradiation on cytokine production both in vitro (using lymphocytes from healthy donors) and in vivo (using serum levels of head and neck cancer patients following irradiation) and correlating them to mucositis severity/need for percutaneous endoscopic gastroscopy (PEG) tube installation.MethodsCytokine production by cultured lymphocytes from healthy donors, in vitro, following irradiation of 5 or 10 Gy. In addition, blood from 23 patients with head and neck cancers, irradiated by 60–72G in vivo, were assessed for inflammatory cytokines (tumor necrosis factor (TNF)α, interleukin (IL)-6, IL-8, IL-18), the anti-inflammatory cytokine IL-10, and the general marker sIL-2R. Following radiation, selected patients who were developing mucositis were treated by PEG tube installation. Changes in cytokine levels were studied as predictive biomarkers of response to therapy/PEG tube installation. Cytokine production levels were measured using ELISAs kits.ResultsIrradiation decreased the levels of all tested cytokines, most notably IL-6 and IL-8, proportional to irradiation dose. In patients, increases in cytokine levels, correlated with mucositis severity and potentially the need for PEG tube installation.ConclusionsIrradiation decreased the levels of all cytokines of healthy lymphocytes in a dose-dependent manner, especially those of IL-6 and IL-8. This study shows a correlation between high and increasing levels of inflammatory cytokines, sIL-2R, plus radiation toxicity and the need for PEG. The reduction of cytokine levels after radiotherapy predicts that PEG will not be required. Thus, our study shows that cytokine changes are predictive biomarkers in head and neck cancer patients.

Uveal melanoma (UM) is the most common primary intraocular malignancy in adults. High serum levels of soluble IL-2 receptor (sIL-2R) have been reported in acute inflammations and metastatic cancers. This study evaluated the potential of high/increasing sIL-2R levels in predicting metastases.The study included a total of 1,546 sera samples of subjects from three groups: 119 healthy controls (73 subjects), 566 UM 10 year (10y) disease-free (DF) (220 patients), 861 metastatic UM (268 patients). Patients were followed-up biannually with liver ultrasound and liver function tests for the presence of metastases (Mets). Blood samples to measure the levels of sIL-2R were obtained at the time of primary diagnosis, soon after initial treatment (enucleation, brachytherapy), every 6 months, 10 years from diagnosis, at Mets confirmation by CT, and after additional treatments.Significantly higher sIL-2R levels were detected in the Mets patients compared to healthy controls and 10y DF patients. Compared to the upper limit of the normal levels of sIL-2R, 1,000 U/ml, its levels in metastatic UM were 61%, 25% in 10y DF UM, and 6.25% in the controls. High levels of sIL-2R in metastatic patients, decreased significantly post treatments. Individual kinetics of markers, indicated similar trends of sIL-2R compared to osteopontin and S-Protein 100, predicting metastases, which were confirmed on liver imaging.Significantly higher sIL-2R levels were evident in all UM patients with Mets. Significant increases in sIL-2R levels on serial evaluations indicated and predicted UM Mets, enabling earlier treatment of Mets, to improve survival.

BACKGROUND: Cancer progression is associated with significant systemic clinical manifestations including cachexia induced weight loss and anorexia. Pentoxifylline (PTX) is a drug that has been shown to have multiple beneficial effects in cancer patients through its anti-inflammatory properties. MAIN OBJECTIVE: To evaluate PTX effects on colon cancer patients treated with chemotherapy. PATIENTS and METHODS: Forty metastatic colon cancer patients receiving chemotherapy were enrolled in this randomized study. 17 patients were treated with a full dose of PTX (400 mg TID), 9 patients with a reduced dose PTX (200 mg TID) and 23 served as controls (no PTX). RESULTS: Follow-up evaluations of patients included the following: physical examination; leukopenia determination; weight determination; stomatitis determination; and survival rate. Patients treated with PTX (both full and reduced doses), experienced a significant increase in weight and a reduction in stomatitis relative to the control group. Treatment with PTX also significantly increased patient survival rate. All patients treated with PTX, had a median overall survival (OS) rate of 20.4 months as compared to 13.2 months in the control group. CONCLUSIONS: PTX treatment of colon cancer patients, in addition to chemotherapy, significantly improved survival rates, induced weight gain and reduced stomatitis occurrence –all important parameters of cachexia.

This article reviews the principal attempts of immune-modulation or immune therapy in metastatic breast cancer. It considers their rationale and reports on results from the relevant key clinical trials. Immune-modulatory or immune-stimulating cytokines used alone or combined with conventional therapies is among the principal approaches of immune manipulation in breast cancer. As this issue has recently been reviewed by us, the aim of the current article is to discuss our updated and unpublished data on this topic. Overall survival in luminal (28 patients) and non-luminal (9 patients) molecular subtypes is 91 and 59 months respectively that is about two and half or three times longer than expected. Thereafter, we focus on monoclonal antibodies (mAb) based-therapies including novel strategies to overcome resistance to anti-HER2 mAb. The main vaccine platforms in different molecular subtypes and immune therapies in triple negative metastatic breast cancer (m-TNBC) are discussed in the last sections. Some phase III investigations have already changed the current clinical practice. In fact, pertuzumab plus trastuzumab and docetaxel is the recommended first line regimen in HER2 positive locally recurrent or metastatic breast cancer and bevacizumab plus paclitaxel or docetaxel is a reasonable option for m-TNBC. In some other observational or phase I/II studies on first-line trastuzumab plus chemotherapy and hormonal therapy and in that on HER2 peptide/protein vaccines promising although preliminary findings have been reported to be further validated. In the remaining studies, results were disappointing. In the future, finding new predictive biomarkers and exploring more suitable synergizing combinations, time and dose-dependent-scheduled sequences of currently and further investigated immunological approaches are main challenges.

Andrea Nicolini,1 Angelo Carpi,2 Paola Ferrari,1 Riccardo Morganti,3 Valentina Mazzotti,3 Vivian Barak,4 Michael J Duffy5 1Department of Oncology, Transplantations and New Technologies in Medicine, University of Pisa, Pisa, Italy; 2Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy; 3Section of Statistics, University Hospital of Pisa, Pisa, Italy; 4Immunology Lab for Tumor Diagnosis, Hadassah University, Jerusalem, Israel; 5Conway Institute of Biomolecular and Biomedical Research, UCD School of Medicine, University College Dublin, Dublin, Ireland Purpose: The purpose of this study was to evaluate the combined measurement of serum CEA, TPA, and CA 15-3, using an individual reference limit (IRL), for predicting distant metastases in asymptomatic women following a diagnosis of primary breast cancer. Methods: A total of 231 patients were followed up for a mean of 5.5±1.6 years. An IRL for defining critical changes (CCs) in marker levels was used as a warning signal of pending distant metastases.Results: Sensitivity, specificity, and accuracy of the combined CEA–TPA–CA 15-3 marker panel for predicting patient outcome were 95.2%, 97.8%, and 97.9%, respectively. In all, 19 (8.3%) patients relapsed with a mean lead time to radiological evidence of metastases of 11.7±13.8 months.Conclusion: We concluded that the combined measurement of CA 15-3, CEA, and TPA using an IRL for determining the CC in markers levels is an accurate strategy for predicting outcome during postoperative monitoring of asymptomatic breast cancer patients. Whether the early prediction of metastasis and subsequent administration of therapy impacts on patient outcome should now be the objective of a prospective clinical trial. The marker panel described here could serve as the basis for such a trial. Keywords: breast cancer, postoperative monitoring, serum tumor markers, individual reference limit, early detection of relapses

Background/aim Prognostic factors serve as a vital tool in the treatment of patients with head and neck cancer (HNC). The aim of this study was to evaluate the clinical potential of Thymidine-kinase-1 (TK1) marker in the prognosis of HNC patients. Patients and methods We evaluated 366 blood samples from 278 HNC patients and 88 healthy controls, using an ELISA assay. Correlations of TK1 levels with disease stage, lymph node involvement and response to radiation therapy, were determined. Results In HNC patients, TK1 levels were significantly higher compared to healthy controls. Significantly higher TK1 levels were demonstrated in node positive cases and in advanced disease stages compared to node negative and early disease stages. Levels were higher prior to radiation and decreased significantly thereafter, in patients responding to treatment. Increasing levels of TK1 post-radiation were indicative of recurrence or of non-response to treatment, while decreasing levels indicated a positive response. Conclusion TK1 is a tumor marker in HNC patients with the ability to assess response to therapy. High or increasing levels correlated to a poor prognosis, whereas low levels correlated to an overall increased survival.

Background/aim There is a need to diagnose early bladder cancer by non-invasive tests. This study aimed to explore the clinical value of three non-invasive methods, UBC Rapid, ultrasound (US), and urine cytology, separately and in combination, for the primary diagnosis and surveillance of bladder-cancer. Patients and methods Urine samples were obtained from 106 patients who presented with symptoms of bladder cancer and patients followed-up after transurethral resection of bladder tumors (TURB). Each patient underwent US, cystoscopy, cytology and UBC Rapid test. The sensitivity and specificity of all methods and combinations were calculated and related to cystoscopy and biopsy. Results Voided urine samples assayed with UBC Rapid and cytology yielded a sensitivity and specificity of 58.3% and 75.9%, and 57.1% and 98.0%, respectively and for US 76.2% and 98.1%. The combination of all three methods resulted in a sensitivity and specificity of 95.8% and 67.3%, and the combination of UBC Rapid and US, gave a sensitivity of 91.3%, and a specificity of 72.2%, The combination of UBC Rapid and cytology yielded a sensitivity and specificity of 84.6% and 71.2%. Conclusion Combined use of UBC Rapid, US and cytology improved the sensitivity of bladder cancer detection.

Several observations indicate that cytokine concentrations might also relate to the severity of the psychosis. In this study we assessed whether inflammatory and anti-inflammatory cytokine concentrations are associated with the degree of the psychotic manifestations. A group of 41 patients with schizophrenia suffering from an acute psychosis leading to hospitalization in a psychiatric ward were assessed for the intensity of their psychotic manifestations by the PANSS score. Serum IL-2R, IL-6, IL-8, IL-10 were analyzed by commercial ELISA kits. These patients were compared to controls without schizophrenia. At the univariate analysis, statistically significant elevated levels of the cytokines IL-6, IL-2R and IL-8 were detected in the sera of the patients with schizophrenia compared to controls. At the multivariate analysis, statistically significance held only for IL-2R concentration. Furthermore, positive correlation was found between symptom severity as measured by the PANSS and IL-6 levels as well as IL-2R levels. In Conclusion, our data indicate that elevated serum concentrations of IL-6, IL-8 and IL-2R are associated with severe clinical symptoms measured by the total, general, negative and positive scores of the PANSS scale.

Mounting data are indicating that major depression is related to diverse functions of the immune system. Several observations indicate that cytokine concentrations might also relate to the intensity of depressive manifestations. In this study we assessed whether inflammatory and anti-inflammatory cytokine concentrations are associated with the intensity of the depressive features in a cohort of patients with major depression and in healthy normal controls.A group of 25 patients with major depression all suffering from an acute deterioration of their mental status and all hospitalized in a psychiatric ward were assessed for the intensity of their depressive manifestations according to the Hamilton rating scale for depression and by the clinical global impression scale (CGI). In parallel, concentrations of serum IL-2R, IL-6, IL-8, IL-10 were analyzed by commercial ELISA kits. As comparators, a group of 25 healthy controls was analyzed.The levels of IL-6 were higher among patients with depression. A high degree of correlation was found between the scores measured by the Hamilton and CGI scales by which the intensity of depressive symptoms were ranked. Interestingly, within the group of patients with depression a negative correlation was detected between the IL-6 concentrations and the CGI scores while a positive correlation was found between the IL-10 concentration and IL-6 concentration.Our data indicate that the patients with depression differ from healthy individuals by their cytokine profile. Within this group of patients depressive features have a specific pattern and linkage to inflammatory and anti-inflammatory scores.

BACKGROUND In vitro studies suggested that circulating inflammatory cytokines cause septic myocardial dysfunction. However, no in vivo clinical study has investigated whether serum inflammatory cytokine concentrations correlate with septic myocardial dysfunction. METHODS Repeated echocardiograms and concurrent serum inflammatory cytokines (IL-1β, IL-6, IL-8, IL-10, IL-18, tumor necrosis factor-α, and monocyte chemoattractant protein-1) and cardiac biomarkers (high-sensitivity [hs] troponin-T and N-terminal pro-B-type natriuretic peptide [NT-proBNP]) were examined in 105 patients with severe sepsis and septic shock. Cytokines and biomarkers were tested for correlations with systolic and diastolic dysfunction, sepsis severity, and mortality. RESULTS Systolic dysfunction defined as reduced left ventricular ejection fraction (LVEF) P = .006 and U tests). In contrast, NT-proBNP strongly correlated with both reduced LVEF and reduced e′-wave velocity, and hs-troponin-T correlated mainly with reduced e′-wave. CONCLUSIONS Unlike cardiac biomarkers, none of the measured inflammatory cytokines correlates with systolic or diastolic myocardial dysfunction in severe sepsis or septic shock.

Objective To evaluate serum levels of interleukin (IL)-1, IL-6, IL-8, IL-10, soluble IL-2 receptor (sIL-2R), squamous cell carcinoma antigen (SCCA), tissue polypeptide–specific antigen (TPS) and vascular endothelial growth factor (VEGF) in patients with potentially malignant disorders (PMD), oral squamous cell carcinoma (OSCC), or status-post (SP) OSCC. Subjects and Methods Blood was collected from 47 patients, either controls or diagnosed with PMD, OSCC, or SPOSCC. Levels of cytokines and tumor marker were evaluated by ELISAs. Normal levels were based on previous studies and pathology determined by chi-square and Fisher's exact tests. P ≤ 0.05 was considered statistically significant. Results Above normal levels of SCCA were found for OSCC and dysplasia patients (33.3% and 11.1%, respectively) and high range of normal (upper 20% of the normal range) for lichen planus, SPOSCC, and dysplasia patients (6.67%, 33.3%, and 22.2%, respectively), differences that approached statistical significance (P = 0.055). No differences were found between groups for other tested markers. A progression was seen for SCCA from high range of normal in SPOSCC to a mixture of high normal and elevated in dysplasia to elevated in active OSCC, suggesting that SCCA may be correlated with cancer progression. Conclusion Higher levels of serum SCCA may serve as a marker for dysplasia and progression to oral carcinogenesis.

Head and neck cancer (HNC) patients are usually diagnosed with advanced disease and multimodality therapies are required, as well as prognostic biomarkers to predict their response and assess survival. In this study, we aimed to evaluate the ability and clinical significance of the immune biomarker sIL-2R in HNC patients, to assess therapy response and prognosis.We evaluated 328 blood samples from 145 head and neck cancer patients (HNC) from several subgroups: 84 larynx carcinomas pre- and 39 post-therapy, 46 oral cavity carcinomas pre- and 29 post-therapy, 12 nasopharynx carcinomas, 16 parotid and other salivary gland carcinoma patients. The control group included 45 healthy subjects. Serum sIL-2R levels were evaluated by ELISA assays and correlated to disease stage, lymph nodes, response to therapy, survival and cancer differentiation.Significantly higher sIL-2R levels were recorded in all HNC patients, as opposed to controls, in advanced versus early-stage disease that decreased following therapy. sIL-2R distinguished best, in comparison to other tumor markers, between HNC patients and controls. Survival was strongly associated to lower sIL-2R levels in patients entering the study.sIL-2R is a sensitive immune marker for HNC patients. Its levels correlate to disease stage, assess response to therapy and are predictive of recurrence or better survival. We suggest, therefore, using sIL-2R as a reliable prognostic marker in HNC patients as a single marker, or in a combined panel of biomarkers.

Objective: Lamininα2-deficient congenital muscular dystrophy type 1A (MDC1A) is a cureless disease associated with severe disability and shortened lifespan. Previous studies have shown reduced fibrosis and restored skeletal muscle remodeling following treatment with losartan, an angiotensin II type I receptor blocker. We therefore evaluated the effect of losartan treatment in the dy2J/dy2J mouse model of MDC1A. Methods: Homozygous dy2J/dy2J and control mice were treated with losartan or placebo for 12 weeks from 6 weeks of age. Outcome measures included hindlimb and forelimb muscle strength by Grip Strength Meter and quantitative muscle fibrosis parameters. Losartan's effects on transforming growth factor β (TGF-β) and mitogen-activated protein kinase (MAPK) signaling pathways were evaluated with Western blotting, immunofluorescence, and cytokine measurements. Results: Losartan treatment was associated with significant impressive improvement in muscle strength and amelioration of fibrosis. Administration of losartan inhibited TGF-β signaling pathway, resulting in decreased serum TGF-β1 level and reduced downstream phosphorylated (P) Smad2/3 proteins. Moreover, losartan activated Smad7 protein, a key negative regulator of TGF-β signaling. In addition, losartan treatment inhibited the MAPK cascade as shown by decreased expression of P-p38 MAPK, P-c-jun-N-terminal kinase, and P-extracellular signal-regulated kinases 1 and 2 in the treated mice. Interpretation: Losartan, a commonly prescribed US Food and Drug Administration-approved medication for hypertension, demonstrated clinical improvement and amelioration of fibrosis in the dy2J/dy2J mouse model of MDC1A via TGF-β and MAPK signaling pathways. The results of this study support pursuing a clinical trial of losartan treatment in children with MDC1A. ANN NEUROL 2012

Malignant melanoma incidence is increasing over the last years, while mortality is strongly decreasing due to improved early detection, close monitoring of patients including disease biomarkers as well as introduction of new therapies. The aim of the present study was to evaluate biomarkers, mainly S-100β in melanoma patients, regarding its ability to assess treatment response, especially to new immunotherapies (anti-BRAF, ipilimumab, anti-PD-1) and evaluation of prognosis of those patients.We evaluated both retrospectively and prospectively 137 malignant melanoma patients. Blood biomarker levels were evaluated by conventional ELISA assays. Correlations of marker levels to disease stage, metastases, response to new immunotherapies and survival, were performed.Serum levels of biomarkers, mainly S-100β, were significantly higher in all patients before various therapies were applied (5.1+0.7 μg/L) and decreased thereafter (1.3+0.4 μg/L). Significantly higher levels of S-100β were demonstrated in advanced disease including metastases, (5.95+0.62 μg/L) as opposed to early disease (0.32+0.06 μg/L) and NED patients (0.18+0.03 μg/L). When comparing melanoma deceased patients who had extremely high levels of S-100β, (2.2+0.45 μg/L) we showed significantly lower levels in alive patients (0.26+0.02 μg/L) and certainly in normal controls (0.07+0.02 μg/L). In individual patients, kinetic evaluations showed earlier response to therapy, or recurrence and non-response, as shown only later by CT evaluations.S-100β can serve as a useful biomarker for the assessment of treatment response and prognosis, especially after using new immunological treatments, such as anti-BRAF, ipilimumab or anti-PD1 in malignant melanoma patients. Additional biomarkers, such as LDH, β2M and TK may also serve as part of a biomarkers panel, for improved detection of recurrence and metastasis of melanoma patients.

Background To determine the kinetics of serum transforming growth factor beta-1 (TGF-β1) in aged hip fracture patients during a month post-op. and determine its relationship to complications and cognitive status. Methods Forty-one elderly patients were prospectively followed. Serum TGF-β1 was obtained during the first 10 h post-fracture (baseline) and pre-surgery, 48–60 h post-op., 7 and 30 days post-op. Results Mean age was 81.8 ± 7.7, range 68–97 years. A significant increase in serum TGF-β1 (from baseline) was observed on the seventh day post-op., lasting for at least 1 month ( p = 0.004). Sixteen patients experienced post-op. complications. Serum TGF-β1 of patients with complications was significantly higher compared to patients without complications ( p = 0.039). More complications were found in impaired mental status (IMS) patients (9/12) compared to cognitively normal patients (10/29), ( p = 0.037). However, no differences were found in serum TGF-β1 between IMS and normal patients, nor was a correlation found between age and TGF-β1 production. No differences were found between genders. Conclusions There is a distinct increase in serum TGF-β1, a week post hip fracture repair, lasting for at least a month, which is significantly higher in patients with post-op. complications. TGF-β1 seems to be essential to the healing process post hip fracture repair; still its clinical applications have as yet to be determined.

We investigated the effect of a mixture of long-chain fructo-oligosaccharides, galacto-oligosaccharides and acidic oligosaccharides on the number and consistency of stools and on immune system biomarkers in 104 supplemented and non-supplemented subjects (aged 9–24 months) with acute diarrhea. Interleukin-1 (IL-1), IL-1RA, IL-6, IL-8, IL-10, TNF-α and sIL-2R cytokine levels were determined. The significant decrease in number of stools and increase in stool consistency in the supplemented group was of little clinical relevance. The only significant change in pro- and anti-inflammatory cytokines was decreased TNF-α levels in the supplemented group. Prebiotic supplementation during acute diarrhea episodes did not influence the clinical course.

Hairy cell leukemia (HCL) is a well recognized indolent B-cell lymphoproliferative disorder. HCL cell proliferation is regulated by growth factors and cytokines, of which tumor necrosis factor-alpha (TNF-alpha) may be one of the most important. The mechanism of TNF-alpha-induced HCL cell growth is mediated via 2 receptors, which are present in both cellular and soluble forms. In this study we determined the serum levels of TNF-alpha and their soluble receptors - sTNF-R60 and sTNF-R80 - in 23 HCL patients and correlated them with clinical parameters before and after therapy. Patients were classified according to their clinical status as either "active" at diagnosis or during relapse and "non-active" (responding to therapy with partial and complete remission). Most patients were treated with 2-CDA, following which serum levels of TNF-alpha, sTNF-R60 and sTNF-R80 were significantly decreased, particularly in CR. Significant differences in paired observation values were noted for TNF-alpha and sTNF-R80, indicating a good correlation with the clinical status of disease, but this was not the case for sTNF-R60 (coefficients of correlation between levels of TNF-alpha and sTNF-R80 and of TNF-alpha and sTNF-R60 were r = 0.85 and r= 0.64, respectively). These results suggest that decreases in both TNF-alpha and sTNF-R80 are indicative of response to treatment, while increased levels accompany active disease. Accordingly, we conclude that serum levels of the TNF family, as for the sIL-2R and IL-1 family, may also be used as sensitive markers for monitoring HCL status. Levels may be indicative of the clinical efficacy of therapy and can be used as an indicator of the presence of residual disease.

A nontoxic dose of Sambucol, an immunomodulator commercially sold as an immune stimulator, was examined in murine models of leishmaniasis and malaria. Sambucol causes a shift in the immune response, as demonstrated in human monocyte cultures, to Th1 (inflammation-associated) responses. Treatment of leishmania-infected mice with Sambucol delayed the development of the disease. As there was no direct IN VITRO anti-leishmanial effect, the observed partial protection IN VIVO is most likely related to immune modulation. Although increased Th1 responses are associated with protection from leishmaniasis, they are considered to be the main immunopathological processes leading to cerebral malaria. Administration of Sambucol to mice prior to and following infection with Plasmodium berghei ANKA increased the incidence of cerebral malaria, while administration of Sambucol after infection had no effect on the disease. The results demonstrate how an inflammatory-like response may alleviate or exacerbate clinical symptoms of disease and hint at the importance of administration timing. The overall effect of immunomodulator administration depends on the ongoing immune response and the Th1/Th2 balance determined by both host and parasite defense mechanisms.

This article analyzes the serum cytokine profile of a nonrandomized group of patients with rheumatoid arthritis (RA) and psoriatic arthritis (PsA) who are destined to be treated with infliximab following failure after failure of different disease-modifiying antirheumatic drugs (DMARDs). Serial serum samples were collected from 11 patients with refractory RA, three with PsA and one with undifferentiated spondyloarthropathy. All were treated with the antitumor necrosis factor (TNF)alpha agent, infliximab, after failing to sustain a clinical remission with conventional DMARDs. Blood samples were obtained at different phases of their therapy. Serum levels of tumor necrosis factor (TNF)alpha, interferon (IFN)gamma, interleukin (IL)-1beta, IL-6, sIL-2R, IL-10, and IL-1 receptor antagonist (IL-1RA) were determined by commercial ELISA kits. Interestingly, only eight of the 11 patients with RA had elevated TNFalpha serum levels (at least once in their serial measurements). Only one was unresponsive to therapy and despite anti-TNFalpha therapy her serum TNFalpha levels remained extremely high. Two RA patients who responded to infliximab had normal TNFalpha serum levels prior to and following infliximab administration. One RA patient improved after infliximab therapy despite unrelenting high serum levels of TNFalpha, IL-6, and sIL-2R. Patients with active PsA who responded to infliximab therapy had sustained high serum TNFalpha levels. In an unselected population of RA and PsA patients, we noticed diverse patterns of serum cytokine profiles. These results imply that the cytokine profiles of RA and PsA are diverse and their pathogenesis is heterogeneous.

Establishing prognostic factors is very important in the management of cancer patients. Our aim was to evaluate the clinical significance of a panel of tumor markers, including CEA (Carcino Embryonic Antigen), SCC (Squamous Cell Carcinoma Antigen), TPS (Tissue Polypeptide Specific Antigen) and CYFRA 21-1 in head and neck cancer patients, for assessing treatment response and prognosis of patients.We evaluated 312 blood samples from 143 head and neck cancer patients, from several sub-groups: 82 Larynx Carcinoma pre- and 38 post-therapy, 46 Oral Cavity pre and 29 post-therapy, 12 nasopharynx, 16 parotid and other salivary gland patients. Blood tumor markers levels were evaluated by conventional ELISA assays. Correlations of marker levels to stage of disease, lymph node involvement and therapy, were performed.Serum levels of all four tumor markers were higher before therapy and decreased thereafter in all patients. The decrease in TPS level following therapy was significant (p=0.03). Significantly higher levels of TPS and similarly higher levels of the other tumor markers were demonstrated in advanced disease (stages III and IV) patients, as opposed to early disease (stages I and II) patients (p=0.012). Node positive patients had significantly higher TPS levels as compared to node negative (p=0.02). The same trend was shown by the other markers as well, but did not reach statistical significance. TPS was best correlated to survival of patients; those having low levels had the best clinical outcome and longer survival.CEA, SCC, TPS and CYFRA 21-1 can all serve as useful tumor markers in HNC patients. They assessed response to therapy and were prognostic for recurrence. TPS proved to be the most sensitive predictor of advanced disease and poor prognosis.

Serum levels of thymidine kinase 1 (TK1), an enzyme involved in the G1-S phase of the cell cycle, have been previously shown to correlate with the prognosis of lymphoid malignancies. We hypothesized that TK1 levels will be higher in aggressive, compared to indolent lymphoproliferative, malignancies and this may serve as a marker of transformation from an indolent to aggressive disease. We analyzed serum from 182 patients and correlated the findings with the type of malignancy and prognosis; we further compared the TK1 levels of 31 patients with a proven transformation and 34 patients with clinically suspected transformation that was eventually deferred. The mean TK1 levels of patients with indolent and aggressive disease was 18.9±3.3 and 39.8±3.3 U/l respectively (p0.001). Among patients with aggressive disease, low TK1 levels correlated with improved survival (p=0.008). TK1 levels16.6 U/l predicted transformation from indolent to aggressive disease (sensitivity of 95%, specificity of 76%, negative predictive value (NPV) of 96% and positive predictive value (PPV) of 69%). A regression analysis showed that only TK1 levels were significant (relative risk (RR)=1.03 for each unit, confidence interval (CI)=1-1.05; p=0.015) for diagnosing a true transformation. In conclusion, TK levels are useful in assessing prognosis, especially in aggressive lymphoproliferative diseases. Moreover, TK levels are adequate in discriminating cases of indolent lymphoma that transformed to an aggressive disease from patients with no proven transformations. This tool provides the clinician a novel method to distinguish between symptomatic patients utilizing a simple test and may lessen the need for aggressive or invasive measures of investigation.

BACKGROUNDBiomarkers are playing increasingly important roles in the detection and management of patients with cancer. Despite an enormous number of publications on cancer biomarkers, few of these biomarkers are in widespread clinical use.CONTENTIn this review, we discuss the key steps in advancing a newly discovered cancer candidate biomarker from pilot studies to clinical application. Four main steps are necessary for a biomarker to reach the clinic: analytical validation of the biomarker assay, clinical validation of the biomarker test, demonstration of clinical value from performance of the biomarker test, and regulatory approval. In addition to these 4 steps, all biomarker studies should be reported in a detailed and transparent manner, using previously published checklists and guidelines. Finally, all biomarker studies relating to demonstration of clinical value should be registered before initiation of the study.SUMMARYApplication of the methodology outlined above should result in a more efficient and effective approach to the development of cancer biomarkers as well as the reporting of cancer biomarker studies. With rigorous application, all stakeholders, and especially patients, would be expected to benefit.

In vitro studies suggested that circulating inflammatory cytokines cause septic myocardial dysfunction. However, no in vivo clinical study has investigated whether serum inflammatory cytokine concentrations correlate with septic myocardial dysfunction.Repeated echocardiograms and concurrent serum inflammatory cytokines (IL-1β, IL-6, IL-8, IL-10, IL-18, tumor necrosis factor-α, and monocyte chemoattractant protein-1) and cardiac biomarkers (high-sensitivity [hs] troponin-T and N-terminal pro-B-type natriuretic peptide [NT-proBNP]) were examined in 105 patients with severe sepsis and septic shock. Cytokines and biomarkers were tested for correlations with systolic and diastolic dysfunction, sepsis severity, and mortality.Systolic dysfunction defined as reduced left ventricular ejection fraction (LVEF)50% or55% and diastolic dysfunction defined as e'-wave8 cm/s on tissue-Doppler imaging (TDI) or E/e'-ratio were found in 13 (12%), 24 (23%), 53 (50%), and 26 (25%) patients, respectively. Forty-four patients (42%) died in-hospital. All cytokines, except IL-1, correlated with Sequential Organ Failure Assessment and APACHE (Acute Physiology and Chronic Health Evaluation) II scores, and all cytokines predicted mortality. IL-10 and IL-18 independently predicted mortality among cytokines (OR = 3.1 and 28.3, P = .006 and0.0001). However, none of the cytokines correlated with LVEF, end-diastolic volume index (EDVI), stroke-volume index (SVI), or s'-wave and e'-wave velocities on TDI (Pearson linear and Spearman rank [ρ] nonlinear correlations). Similarly, no differences were found in cytokine concentrations between patients dichotomized to high vs low LVEF, EDVI, SVI, s'-wave, or e'-wave (Mann-Whitney U tests). In contrast, NT-proBNP strongly correlated with both reduced LVEF and reduced e'-wave velocity, and hs-troponin-T correlated mainly with reduced e'-wave.Unlike cardiac biomarkers, none of the measured inflammatory cytokines correlates with systolic or diastolic myocardial dysfunction in severe sepsis or septic shock.

The use of biomarkers in oncology has been much more extensive than in any other diseases for more than 30 years. Their use has evolved beyond the research laboratory to more routine clinical use as in diagnosis, monitoring of disease status, and treatment efficacy. With the advances in molecular biology, other biomarkers than the “classical” serum markers are being developed for clinical use: gene mutations, microRNA, circulating tumor cells, circulating DNA, and others currently in research. Biomarkers such as these are currently being used for prediction: potential “at risk” for disease as well as response to therapy (predictive biomarkers), and for disease prognosis (prognostic biomarkers). In oncological routine, the correlation of biomarker levels with the clinical status and medical imaging methods is of great significance. The purpose of the correlation is twofold: providing sequential and/or more specific diagnostics and keeping these procedures within affordable economical limits. This special issue includes a number of reviews and research papers dealing with the above mentioned topics. Reviews on Novel Biomarkers. The advances in molecular research and various “–omics” techniques have enabled us to study a number of potential novel cancer biomarkers. Studying gene expressions and molecular pathways may lead to the discovery of new prognostic biomarkers, markers of early diagnostics, or molecular targets for therapy. E. Lastraioli et al. present a review on the expression of hERG1 potassium channels in different types of solid cancer (including breast, esophageal, gastric, colorectal, pancreatic, and other cancers). The authors, on the basis of publication review as well as their own findings, argue that hERG1 could be a novel biomarker and point out the overexpression of hERG1 in solid cancers, a feasible determination by immunohistochemistry and the potential of using them as therapy targets as the monoclonal antibody to block them is already available. M. Rihacek et al. review B-cell activating factor (BAFF), a transmembrane protein, as a biomarker of malignant disease activity and prognostic factor in B-cell derived malignancies such as multiple myeloma. The authors report its proinflammatory properties and its contribution to cancer cachexia. Moreover, BAFF/BAFF-R signaling may be a promising target of future therapy in B-cell derived leukemias and lymphomas. Cytoglobin, a protein of the globin family, is reviewed by T. Bholah et al. Thanks to the advances in molecular research, a possible role of this protein in cancer has been suggested. In their paper, the authors overview different functions of cytoglobin and provide a perspective on potential research areas that may elucidate its role as cancer biomarker. The gene for gamma-glutamylcyclotransferase, an enzyme involved in glutathione metabolism, has recently been investigated and reported as an upregulated protein in various cancers. S. Kageyama et al. report an overview of the activities of GGCT in cancer cells. Imaging Methods and Oncology. Imaging methods provide a useful tool in cancer diagnostics and management. Their applications develop in reflection to new technologies and advances in molecular research. M. Tolia et al. report the value of preirradiated in vivo magnetic resonance spectroscopy parameters in predicting recurrence free survival for patients with high grade gliomas, who undergone postoperative radiotherapy. R. Fusco et al. present the evaluation of the diagnostic value of an imaging protocol that combines dynamic contrast-enhanced MRI (DCE-MRI) and diffusion-weighted imaging (DWI) in patients with suspicious breast lesions. In addition, further research sought to determine if additional information provided by DWI could improve the diagnostic value of breast MRI. Predictive Biomarkers and Risk Factors. J. H. Kim and S. K. Hong report a review, based on meta-analysis of PubMed publications, on potential novel biomarkers in active surveillance of low-risk prostate cancer. These include %[-2] proPSA and Prostate Health Index (PHI), PCA3, TMPRSS2:ERG, the genomic prostate score (GPS), a panel of four kallikrein markers, and the expression levels of different cell cycle progression (CCP) genes. Y. Cao et al. review a meta-analysis of 25 independent epidemiological studies on the association between hormonal and reproductive factors and thyroid cancer risk. As the title of their paper suggests, reproductive factors but not hormonal factors are associated with thyroid cancer risk. R. V. Liubota et al. report risk factors of the invasive breast cancer locoregional recurrence. The authors discovered that the scope of the surgical intervention (breast-conserving surgery (BCS) or radical mastectomy (RME)) does not essentially affect the recurrence appearance frequency or the recurrence-free period duration. However, in the BCS group, risk factors such as the presence of metastases in the regional lymph nodes or the hyperexpression Her/2neu presence increased the frequency of the locoregional breast cancer recurrence appearance. Prognostic Biomarkers. M. Tolia et al. describe a study which aimed to identify whether or not the expression of serum baseline C-reactive protein (CRP) and albumin are related to overall survival in non-small-cell lung cancer (NSCLC). The study results suggested that, in fact, high pretreatment CRP and low albumin serum levels were promising independent prognostic factors of overall survival in NSCLC. Biomarkers in Treatment Monitoring. Several indicators, including tumor markers, are used to monitor whether or not a particular cancer treatment is efficient with tolerable toxicity for patients. However, if a tumor marker has low sensitivity and/or specificity, adding another biomarker to the regimen could potentially enhance the treatment evaluation. S. Kristiansen et al. present a study that focused on the association between hypermethylated DNA and the tumor markers CA 15-3, CEA, and TPA in serum during monitoring of patients with advanced breast cancer. Disease Monitoring and Management. A combination of biomarkers or a set of tests provide a higher sensitivity and/or specificity than a unique biomarker or test. A. Pouliakis et al. report the evaluation of classification and regression trees for the triage of women at risk for cervical intraepithelial neoplasia. The computer-assisted algorithm was based on cytological HPV DNA typing, HPV mRNA detection, p16 immunocytochemical expression, and age and parous status. The authors proposed a methodology which could dramatically reduce the number of women that would require a colposcopy. This special issue aims to prove the importance of biomarkers for the individualisation of the approach to an oncological patient and the improvement of his quality of life. Biomarkers in oncology are an important tool for diagnostics, prognosis, and therapy monitoring on condition that there is a clear goal for the biomarkers determination; biomarkers are monitored systematically and are interpreted in collaboration of the laboratory and the clinician. Single shot biomarker determination is useless and can lead to incorrect assumptions. Marie Karlikova Ondrej Topolcan Olive T. J. Wolfe Vivian Barak Tomas Zima

Chromogranin A (CGA), Pro-gastrin-releasing peptide (ProGRP) and neuron-specific enolase (NSE) are known as immunohistochemical tissue markers closely associated with neuroendocrine differentiation in non-small-cell lung carcinoma (NSCLC). The aim of the present study was to assess the value of serum levels of these markers in predicting response to chemotherapy and survival of patients with unresectable NSCLC.The study included 67 patients with advanced NSCLC treated with chemotherapy. Before treatment, serum levels of CGA, ProGRP and NSE were measured with commercial kits.No association was found between serum NSE and age, gender, histology, performance status or extent of the disease. Distribution of serum CGA differed significantly according to gender and histology, with higher levels being found in men (p = 0.01) and in squamous cell carcinoma (p = 0.01). Serum ProGRP levels correlated with disease extent, being higher in patients with metastatic disease (M1) than in those with locoregional disease (M0; p = 0.02). The association of NSE, CGA and ProGRP levels with response to chemotherapy was not significant. While NSE had no impact on survival, the median survival was shorter for patients with elevated serum CGA and longer for patients with high ProGRP levels. Association with survival was significant when the Classification and Regression Tree (CART)-derived or median cutoff points were explored. On inclusion in multivariate Cox models, both CGA and ProGRP retained significance with high levels showing an opposite effect on survival [CART-derived cutoff points: CGA, relative risk (RR) -4.0; p0.001, and ProGRP, RR -0.4; p = 0.006, and median cutoff points: CGA, RR -1.8; p = 0.04, and ProGRP, RR -0.5; p = 0.03]. The combined use of CGA, ProGRP and NSE allowed for definition of two sets of patients with significantly different median survival times (25.2 vs. 8.8 months, p = 0.0001).In the circulation, CGA and Pro-GRP appear to bear important information related to the prognosis for NSCLC patients before chemotherapy. While a high CGA before treatment was found as an unfavorable prognostic determinant, a high ProGRP conferred a survival advantage. The combined use of serum CGA, ProGRP and NSE may supply additional information to prognosis.

Reducing the blood supply of tumors is one modality to combat cancer. The objective of this study was to evaluate such an approach in the treatment of localized murine AML (acute myelogenous leukemia). For this purpose we designed an experimental model in which leukemic cells were embedded in 1% agar discs before subcutaneous implantation in C57Bl female mice. The C-1498 AML cell line (Frederick Inst., NCI, MD, USA) was used. Thirty experimental mice received on alternate days injections of 5 x 2.5 microg anti-VEGF (vascular endothelial growth factor) and 5 x 2.5 microg anti-Flk-1 (VEGFR2) antibodies to the site of cell implantation over a period of 10 d. Fifteen control mice received daily PBS injections. All mice were sacrificed 16 d after AML implantation. Of the 30 experimental animals, macroscopic examination showed in 21 animals (70%) small sized, pale tumors (0.5 g); in six mice (20%) the tumors were replaced completely by necrotic tissue, while in three mice (10%), there were large (2.5 g), highly vascularized tumors. In all 15 control mice large highly vascularized tumors were seen. A separate group of mice was studied for total survival following AML implantation. While 12 mice in the control group not treated with antibodies survived for 16 d post-implantation, survival was prolonged in 15 antibody treated mice by approximate 30 d to a total survival time of 48 d. Tumor specimens were processed for histology, immunohistochemistry (IHC) for CD31 endothelial cell antigen, and tube-like formation assay. The small, pale tumors of antibody treated animals consisted of degenerate hyaline material with remnant nests of leukemic cells, whereas large tumors showed sheets of leukemic cells and numerous blood vessels. Specimens processed for CD31 antigen showed scarce or absence of blood vessels in the small, pale tumors in contrast to intensive staining from a rich network of blood vessels in the large, highly vascularized tumors. Tube-like formation assays disclosed rudimentary Grade 1 endothelial cell tubes in the small, pale tumors as opposed to polygonal Grade 4 tube formation in control animals. In conclusion, this murine model of localized AML allows assessment of anti-angiogenic tumor regression. Anti-angiogenic antibodies against VEGF and Flk-1 have therapeutic effects in murine AML.

We report a 13-year-old boy with refractory nephrotic syndrome (minimal change with mesangial proliferation) who failed the standard treatment protocols. There was some temporary response to large steroid doses, but even the Mendoza protocol could not induce remission. We show suppression of the proteinuria with Infliximab (Remicade) with tapering of steroids. Serial serum levels of tumor necrosis factor (TNF)-alpha are shown and discussed. We suggest studying the TNF-alpha blocking agents as optional treatment for nephrotic syndrome.

Objective To assess the involvement of interleukin-18 (IL-18) in the pathophysiology of severe ovarian hyperstimulation syndrome (OHSS), and study its use as a marker of disease and its correlation to capillary hyperpermeability. Design Prospective controlled study. Setting An IVF unit in a tertiary medical center. Patient(s) Twenty-four patients with OHSS in an IVF program, two control groups: group 1, 40 healthy age-matched women without ovulation-induction treatment; group 2, 19 women who received the same ovulation-induction regimen without experiencing OHSS. Intervention(s) Blood samples were obtained at three times: during acute OHSS, on significant clinical improvement, after complete resolution. Ascitic and pleural fluids were obtained by therapeutic paracentesis. Serum, peritoneal, and pleural fluids were analyzed for IL-18 and IL-6, and blood for hematocrit, white blood cell count, and E 2 levels. Main outcome measure(s) Hematocrit white blood cell count, serum, peritoneal, pleural fluid levels of IL-18, IL-6, E 2 in severe OHSS. Result(s) Significantly higher IL-18 levels were detected in serum, peritoneal, and pleural fluids of patients with severe OHSS as compared with both control groups. Serum IL-18 dropped significantly on transition to the diuretic phase and resolution. A statistically significant correlation between serum IL-18 and hyperpermeability characteristics (white blood cell count, hematocrit), serum E 2 , and IL-6 levels was recorded. Conclusion(s) This is the first study suggesting a role of IL-18 as a marker of OHSS, with correlation to capillary hyperpermeability parameters.

Background. The role of inflammatory cytokines in the pathophysiology of arrested labor is currently unknown. We measured levels of inflammatory cytokines in maternal serum and peritoneal washings in arrested first stage of labor to assess the possible involvement of these mediators in this obstetric disorder. Methods. This was a prospective, case-control study involving 15 women who underwent cesarean section for arrested cervical dilatation (group I), 15 controls who were operated during active labor for nonreassuring fetal heart-rate tracing (group II) and 15 controls who were operated electively (group III). Blood samples were drawn from all women shortly before the operation. The presence of peritoneal fluid was assessed and peritoneal washings were obtained during the operation. All samples were assayed for the inflammatory cytokines interleukin (IL)-1β, IL-6, IL-8 and soluble IL-2 receptor (sIL-2R) by solid-phase enzyme-linked immunosorbent assay (ELISA). Results. All women from group I and one pat...

BACKGROUND Halofuginone, an inhibitor of collagen type I synthesis, is an anti-angiogenic agent. Here we evaluated the efficacy of halofuginone to inhibit prostate cancer (PC) xenografts representing various phenotypes of the disease. METHODS An androgen-dependent (CWR22), an androgen-independent (PC3), and a neuroendocrine (WISH-PC2) PC xenograft were used. Halofuginone was given orally or injected intraperitoneally. Tumor size, collagen α1(I) gene expression (in situ hybridization), collagen content (sirius red staining), angiogenesis (immunohistochemistry with factor VIII antibodies), and apoptosis/necrosis (DNA fragmentation) were evaluated. RESULTS Halofuginone inhibited the growth of all subcutaneously implanted xenografts and of WISH-PC2 when transplanted orthotopically. The effect was dose-dependent (WISH-PC2) and accompanied by decrease in plasma PSA levels (CWR22). In all xenografts, halofuginone inhibited collagen α1(I) gene expression, reduced collagen content, and endothelial cell number resulting in an increase in apoptosis/necrotsis. CONCLUSIONS Oral administration of halofuginone slowed the progression of PC xenografts representing a broad range of phenotypes. Halofuginone may become a new modality for PC prevention. Prostate 51: 73–83, 2002. © 2002 Wiley-Liss, Inc.

Background: Heparin and low molecular weight heparin (LMWH) derivatives are considered angiogenic factors in ischemic and infarcted myocardium. However, the effect of dalteparin sodium (fragmin®, a LMWH derivative) on angiogenesis post infarction has not yet been investigated.

Dysregulation of the cytokine network plays an important role in graft-versus-host disease (GVHD). Interleukin-18 (IL-18) is an obligatory cytokine for interferon-gamma (IFN-gamma) production and IFN-gamma and sIFN-gammaR are elevated in patients with GVHD. Because IL-18 binding protein (IL-18BP) is an inhibitor of IL-18-mediated IFN-gamma production, we evaluated IL-18BP levels in patients undergoing allogeneic peripheral blood stem cell transplantation (PBSCT). IL-18BP levels were assessed in 14 patients on day -10 (before conditioning), on the day of transplant, on the day of engraftment, and during transplant-related complications. A comparison of the kinetics of IL-18BP and soluble(s) IL-6R, sIFN-gammaR, IL-18 serum levels was performed. IL-18BP levels were assessed by specific monoclonal antibodies in a double-sandwich enzyme-linked immunosorbent assay (ELISA). In all patients IL-18BP levels decreased during conditioning and increased in parallel with engraftment (p0.05). Accordingly, during rejection, IL-18BP serum levels remained low and similar to pretransplant levels. The mean elevation of IL-18BP detected in association to acute GVHD was significantly higher in comparison to normal engraftment (p0.05). A correlation between IL-18BP, sIFNgammaR, and sIL-6R serum levels was found in all patients. No correlation between IL-18 and IL-18BP serum levels was found in patients undergoing uneventful PBSCT and rejection, whereas a marked increase in both IL-18 and IL-18BP levels was detected during acute GVHD (p0.01). Our data suggest that the dysregulation of IL-18 and IL-18BP may be important in the pathophysiology of transplant-related complications. Furthermore, because preliminary data from our group show that IL-18 blockage ameliorates GVHD in murine models, it is inferred that these cytokines may represent potential targets in the development of new therapeutic strategies in acute GVHD.

OBJECTIVE The aim of the study was to investigate ex-vivo the bone marrow (BM) stroma of mice under conditions of low- and high-dose cytosine arabinoside (Ara-C), a cycle-specific drug (S-phase) and to assess possible stromal damage, apart from the killing of hematopoietic cells. Stroma consists of mesenchymal elements generally not in the cell cycle; therefore it could not be a target for the killing effect of Ara- C. MATERIALS AND METHODS The stromal function was studied by the following: the incidence of stromal stem cells, i.e. CFU-F; formation of stromal layers under growth conditions of long-term culture (LTC) followed by irradiation and overlayering of test cells in contact and non-contact co-cultures; subsequent culture of the test cells in a semi-solid medium to assay the incidence of hyperproliferative potential cells (HPPC); production of GM-CSF, IL-3, IL-4, IL-6 and IFNgamma in the conditioned medium (CM) of confluent stromal layers. All tests and assays were carried out on BM specimens, 1-4 d after Ara-C administration and on controls. RESULTS Low-dose Ara-C induces a marked decrease of CFU-F, compensated by cycle induction of pre-CFU-F, young-type stromal stem cells. High-dose Ara-C causes a CFU-F decrease to almost zero level. The time length to layer confluency is normal after low-dose Ara-C ( approximately 10 d) and prolonged after a high dose ( approximately 30 d). The confluent layers from mice receiving low- or high-dose Ara-C support hematopoiesis adequately. Among the growth factors and cytokines assayed, only IL-6 is detected in CM layers. IL-6 decreases after a low dose of Ara-C and increases after a high dose. The cause of IL-6 fluctuations is yet to be investigated. It is, however, evident that IL-6 is not an essential factor in support of hematopoiesis. CONCLUSIONS Taken together, the current study in mice indicates that Ara-C administration, in particular a high dose, induces bone marrow stromal damage and/or disfunction. The long period of time to reach layer confluency after a high Ara-C dose might reflect the in-vivo situation of slow stromal regeneration.

BackgroundLow serum vitamin D concentrations have been reported in several autoimmune disorders.ObjectiveTo assess whether low serum vitamin D concentrations are related to disease activity of patients with systemic lupus erythematosus (SLE).Methods378 patients from several European and Israeli cohorts were pooled and their disease activity was measured by two different methods: 278 patients had SLE disease activity-2000 (SLEDAI-2K) scores and 100 patients had European Consensus Lupus Activity Measurement (ECLAM) scores. In order to combine the two systems the scores were converted into standardised values (z-scores), enabling univariate summary statistics for the two variables (SLEDAI-2K and ECLAM). The commercial kit, LIAISON 25-OH vitamin D assay (310900-Diasorin) was used to measure serum concentration of 25-OH vitamin D in 378 patients with SLE.ResultsA significant negative correlation was demonstrated between the serum concentration of vitamin D and the standardised values (z-scores) of disease activity scores as measured by the SLEDAI-2K and ECLAM scales (Pearson's correlation coefficient r=−0.12, p=0.018).ConclusionsIn a cohort of patients with SLE originating from Israel and Europe vitamin D serum concentrations were found to be inversely related to disease activity.

Objective: To investigate the effect of ventilation with 100% oxygen on lung injury associated with surgery involving cardiopulmonary bypass (CPB). Design: A prospective randomized study. Setting: University hospital. Participants: Thirty patients undergoing coronary artery bypass graft surgery with CPB. Interventions: Patients were randomized to receive 100% oxygen (Oxygen group) or 50% oxygen (Air group) throughout surgery. During CPB, patients' lungs in the Air group were flushed with air and in the Oxygen group with 100% oxygen. Measurements and Main Results: Lung injury was evaluated by arterial oxygen tension-inspired oxygen concentration (PaO 2 -F I O 2 ) ratio and cytokine levels (tumor necrosis factor-α and interleukin-8) in blood and bronchoalveolar lavage fluid measured before and after CPB. The lowest PaO 2 -F I O 2 value was observed after 40 minutes following the completion of CPB in both groups. PaO 2 -F I O 2 values 6 hours after CPB were not different from baseline in the Air group but remained lower (359 ± 63 mmHg and 298 ± 78 mmHg; p = 0.013) in the Oxygen group. Blood cytokine levels rose during surgery in both groups. Bronchoalveolar lavage levels of interleukin-8 did not change, whereas tumor necrosis factor-α increased only in the Oxygen group ( p = 0.035). Conclusions: A significant decrease of oxygenation was observed in the early post-CPB period in both groups of patients, with delay in recovery in patients treated with 100% oxygen. A larger increase of the proinflammatory cytokines was found in patients treated with 100% oxygen. High oxygen concentrations during surgery with CPB should be used only when specifically required. Copyright © 2000 by W.B.Saunders Company

Spontaneous fluctuations in activity of low-grade B cell lymphomas are common but not understood. An explanation may be offered by studying an atypical SLL/CLL case characterized by recurrent cycles of leukemic phase alternating with spontaneous remission (1). During remissions, residual IgMkappa+ leukemic cells exhibited resting phenotype, low proliferative response to CD4O-ligand and delayed apoptosis. In contrast, the acute phase counterparts were phenotypically activated, underwent rapid apoptosis in culture and proliferated extensively in response to membrane-anchored CD40-ligand. Transient bursts of serum TNFalpha and IL-10 preceded the acute phases, which were characterized by the co-existence of CD40-ligand+ T lymphocytes and lymphoma cells in the bone marrow. Based on ex-vivo and in-vitro data, we suggest that changes in the lymphoma milieu affect the neoplastic cell activation status, rate of proliferation in response to activated T cells and rate of apoptosis. These responses may underlie both the induction and spontaneous regression of the acute phases in this unique lymphoma. Our findings raise the possibility that part of this mechanism may have evolved during transformation of indolent common CLL to its more aggressive form.

Thirteen patients with DSM-III-R diagnosis of either major depression or bipolar I depression participated in the study. The control group consisted of 10 subjects evaluated for headache or suspected meningitis, none of whom were found to suffer from any organic disease. CSF was withdrawn from all subjects for the measurement of soluble interleukin 2 receptor (sIL-2R). CSF sIL-2R levels were found to be lower in patients as compared to controls (df = 1, 20; F = 84; p < 0.000001).

SUMMARYMice with experimental anti-phospholipid syndrome (APS), induced by active immunization with a human anti-cardiolipin MoAb (H-3), were treated with mouse anti-idiotypic MoAb (anti-H3, named S2.9) and with an irrelevant anti-idiotype. The immunized mice produced high titres of mouse anti-cardiolipin antibodies along with clinical manifestations of experimental APS: prolonged activated partial thromboplastin time (aPTT), thrombocytopenia and high rate of fetal loss. Treatment with the specific anti-Id (S2.9) as a whole molecule or F(ab)2 fraction, resulted in a decrease in serum levels of the anti-cardiolipin antibodies, rise in platelet count, shortened aPTT and reduced rate of fetal loss. The anti-Id effect was associated with a rise in the number of IL-2 and interferon-gamma (IFN-γ)-secreting cells (Th1) and reduction in IL-4- and IL-6-secreting cells (Th2). The beneficial effect of the anti-Id treatment in mice with experimental APS induced by active immunization with an idiotype further supports the idiotypic aetiology of experimental APS and points to the role of Th1 cytokines in suppression of its manifestations.

Cutaneous T-cell lymphomas (CTCL) are diseases characterized by cutaneous infiltrates of malignant clonally expanded T cells. CTCL cells exhibit a cytokine profile consistent with T helper-2 type (TH2) cells. Eosinophilic pneumonias are individual syndromes characterized by eosinophilic pulmonary infiltrates and commonly peripheral blood eosinophilia. CTCL and chronic eosinophilic pneumonia are rare clinical entities. We report a patient with the association of CTCL and chronic eosinophilic pneumonia. To understand the mechanism leading to the eosinophilia, we examined the patient's cytokine profile. This was consistent with a high TH2 activity. Her interleukin (IL) 5, 6, and 10 levels were extremely high, while her IL-2 and interferon-gamma (IFN-gamma) levels (TH1 profile) were low. We believe that eosinophilic pneumonia in this patient is probably secondary to high TH2 cytokine levels induced by tumor cells. We suggest that eosinophilic pneumonia should be considered as a possible diagnosis in patients with CTCL who have respiratory complaints.

Oral tolerance was induced in BALB/c mice by feeding low dose β2-glycoprotein I (β2GPI). The β2GPI-fed mice did not develop serologic and clinical markers of experimental antiphospholipid syndrome (APS) upon immunization with the autoantigen. The treated group was characterized by low titers of serum anti-β2GPI and anticardiolipin Abs in the serum, lack of fetal resorptions, low incidence of thrombocytopenia, and normal aPTT (activated partial thromboplastin time) values. β2GPI given orally before priming with β2GPI resulted in complete prevention of experimental APS development; β2GPI given at an early stage of the disease reduced clinical manifestations. However, administration of β2GPI 70 days postimmunization had a less significant effect on disease expression. Tolerized mice exhibited a diminished T lymphocyte proliferation response to β2GPI in comparison with β2GPI-immunized mice fed with OVA. When nontolerant β2GPI-primed T lymphocytes were mixed with T lymphocytes derived from tolerized mice, a significant inhibition of proliferation upon exposure to β2GPI was observed. The induction of suppression was β2GPI specific and driven, as well as TGF-β mediated. The β2GPI-specific response of T lymphocytes from the β2GPI-fed mice was reversed by anti-TGF-β Abs. The tolerance was adoptively transferred by CD8+ T cells from the tolerized mice into naive mice. Those CD8+ cells were MHC class I restricted, found to secrete TGF-β, and had no cytolytic activity. Oral administration of β2GPI suppressed priming of CTLs in the recipient mice. In sum, β2GPI-induced oral tolerance has an immunomodulatory effect in experimental APS, demonstrating the importance of β2GPI in the pathogenesis of the disease.

BACKGROUND: Sepsis occurs following the presence of bacteria in the circulation and is associated with fever, hyperthermia, and hypotension. Hypophosphatemia develops in the early stages of sepsis. High levels of inflammatory cytokines also characterize early sepsis. AIM: The aim of the present study was to correlate hypophosphatemia with cytokines and cytokine receptor levels during early sepsis. We aimed to reestablish the results obtained from patients in an in vivo experimental model, in order to understand the mechanism of hypophosphatemia induction in early sepsis. METHODS: Ninety-nine patients were enrolled in this study and their clinical condition was classified as the presence of infection, sepsis, and bacterial growth in blood cultures. Phosphate levels and cytokine levels were recorded. In order to determine whether hypophosphatemia is correlated to the increased inflammatory cytokines, we injected normal mice with recombinant cytokines and studied their effect on phosphate levels. RESULTS: Our results revealed that 80% of the septic patients had hypophosphatemia associated with very high levels of tumor necrosis factor (TNF)α and interleukin (IL)-6 and of soluble IL receptor (sIL)-2R and IL-6R, especially in those patients with positive blood cultures. Injection of IL-6, TNFα and IL-1β in mice markedly decreased the phosphate serum levels. CONCLUSIONS: Significant associations were demonstrated between high levels of inflammatory cytokines and their receptors and between serum phosphate levels, especially in patients with positive blood culture. Our results point to a correlation between the high inflammatory cytokines levels and hypophosphatemia during early sepsis. Cytokine levels and hypophosphatemia may be included in sepsis evaluation and prognosis. Anticytokine strategies might, therefore, reverse hypophosphatemia and other parameters of sepsis.

Lipopolysaccharide (LPS) is the major component of the outermost membrane of Gram-negative bacteria and is considered to be one of the major virulence factors of these bacteria. While the effect of systemic injection of LPS is well characterized, the characterization of cytokine secretion in response to local injection of LPS is lacking. The present study was designed to determine the local production of tumor necrosis factor α (TNFα) and interleukin-1β (IL-1β) over a 4 day period following injection of LPS into subcutaneous implanted chambers in mice. Mice were challenged by a single or repeated injection of Salmonella typhosa LPS into the chambers. Chamber fluids were aspirated at different time intervals and were used for assessment of leukocyte and cytokine levels. A single injection of LPS was found to induce cell influx into the chamber which peaked after 4 h. TNFα and IL-1β levels increased rapidly, reaching their maximum levels within 4 h. After 24 h, TNFα levels declined markedly and were undetectable at 48 and 96 h. TNFα mRNA levels in the sedimented cells followed a similar pattern. In contrast, IL-1β showed a more gradual decrease with levels significantly different from baseline still being present 96 h post-LPS challenge. Four consecutive daily injections of LPS into the chambers resulted in undetectable levels of TNFα in the chamber fluid, while significant levels of IL-1β were detected. These levels were significantly higher than the levels of IL-1β in the chamber fluid 96 h after a single injection and approximately 60% of the levels measured 24 h after a single intra-chamber injection of LPS. The results emphasize the difference between single and repeated exposure to LPS in vivo, and suggest a role for TNFα in the initial phase of the local inflammatory response and for IL-1β in the later phase.

Purpose To determine the levels of interleukin-1 receptor antagonist (IL-1ra) in the blood serum of patients with idiopathic bilateral pars planitis and Behcet disease. Methods Five milliliters of blood was withdrawn from the cubital vein of 91 patients (58 with the ocular type and five with the combined type of Behcet disease; 28 with pars planitis) and 36 volunteers. Serum was separated from these samples and stored at – 70 C until assayed. Interleukin-1 receptor antagonist levels were determined by human IL-1ra enzyme-linked immunosorbent assay kits. In patients not receiving any systemic medication, one serum sample was obtained before initiating treatment and another when the patients had been under full medical treatment for 6 weeks or more. Results Pretreatment mean ± SD serum IL-1ra levels were 320 ± 32 pg/ml for the patients with pars planitis, 380 ± 54 pg/ml for patients with Behcet disease, and 271 ± 29 pg/ml for the control subjects (no statistical significance). During treatment, a mean serum level of 352 ± 37 pg/ml was observed in patients with pars planitis (not significantly different from control subjects) and 538 ± 79 pg/ml in patients with ocular Behcet disease ( P = .0116 compared with control subjects). The greatest increase in IL-1ra levels was observed in patients with Behcet disease who received a combination of cyclosporine and corticosteroids. Conclusions Because IL-1ra is one of the natural immunomodulating molecules, the significant increase of serum IL-1ra levels, especially after combined treatment with cyclosporine and corticosteroids, could indicate that the therapeutic effects of this regimen may be mediated through its effects on this molecule.

Myocardial infarction (MI) is the most common cause of cardiac injury in the Western world. Cardiac injury activates innate immune mechanisms initiating an inflammatory reaction. Inflammatory cytokines and vascular cell adhesion molecules (VCAM) promote adhesive interactions between leukocytes and endothelial cells, resulting in the transmigration of inflammatory cells into the site of injury. Low vitamin D levels are associated with higher prevalence of cardiovascular risk factors and a higher risk of MI. In this paper, we examine the effects of short-term vitamin D supplementation on inflammatory cytokine levels after an acute coronary syndrome. We recruited patients arriving to the hospital with an acute MI. All patients received optimal medical therapy and underwent a coronary catheterization. Half of the patients were randomly selected and treated with a daily supplement of vitamin D (4,000 IU) for 5 days. A short course of treatment with vitamin D effectively attenuated the increase in circulating levels of inflammatory cytokines after an acute coronary event. Control group patients had increased cytokine and cellular adhesion molecules serum concentrations after 5 days, while the vitamin D-treated group had an attenuated elevation or a reduction of these parameters. There were significant differences in VCAM-1 levels, C-reactive protein, and interleukin-6. There were trends toward significance in interleukin-8 levels. There were no significant differences in circulating levels of intercellular adhesion molecule 1, E-selectin, vascular endothelial growth factor, and tumor necrosis factor-α. These findings provide information on the anti-inflammatory effects of vitamin D on the vascular system and suggest mechanisms that mediate some of its cardioprotective properties. There is place for further studies involving prolonged vitamin D treatment in patients suffering from ischemic heart disease.

We investigated the in vitro antitumor activity of monocytes derived from autologous bone marrow transplanted (ABMT) patients treated in vivo with granulocyte-macrophage colony-stimulating factor (GM-CSF). Thirty-four patients (17 female, 17 male), median age 42 (range 3-57) years, were enrolled in the study. Fourteen patients were diagnosed with non-Hodgkin's lymphoma (NHL), eight with Hodgkin's disease (HD), nine with breast cancer and three with neuroblastoma. Six patients who did not receive GM-CSF post-ABMT served as controls. We assessed cytotoxicity, antibody-dependent cellular cytotoxicity (ADCC), expression of the activation antigen CD16, and cytokine production by an enriched population of monocytes (90% CD+14) pre-, during and post-GM-CSF administration. Within the group of patients receiving treatment, ADCC was significantly higher during in vivo GM-CSF administration than post-therapy (P0.05) and in 50% of these patients, ADCC increased during in vivo GM-CSF administration over pretreatment values. In addition, in vivo GM-CSF administration caused the monocytes to secrete elevated levels of tumor necrosis factor-alpha (TNF-alpha) and GM-CSF (P0.05). We conclude that GM-CSF augments monocyte-mediated cytotoxicity post-ABMT, and therefore may have a role in controlling minimal residual disease post-transplant.

The objective of this study was to examine the age-dependent relationships between levels of inflammatory cytokines and collagen in human gingival inflammation. The gingival biopsies were obtained from 142 patients, divided into the following age groups: 6 to 14 years (prepubertal children); 18 to 35 years (young adults); 36 to 54 years (mature adults); and 55 years or above. The patients were also divided according to the severity of gingivitis. The tissues were analyzed for the contents of the inflammatory cytokines interleukin (IL)-1 beta, IL-6, IL-8, and tumor necrosis factor alpha (TNF-alpha) using specific ELISA kits, and interstitial collagen type I and type III using the ELISA method and specific antibodies. We found that in young adults, levels of IL-1 beta and IL-6 were significantly higher in inflamed than in non-inflamed gingiva. Total collagen in the young adults, however, was lower in inflamed than in non-inflamed gingiva. There was no significant difference in the levels of either IL-8 or TNF-alpha between inflamed and non-inflamed gingiva independent of age. No difference in the level of collagen type I between the inflamed and non-inflamed gingiva was found in any age groups. The level of collagen type III was lower in inflamed than in non-inflamed gingiva in both children and > or = 55 year group. The results indicate a disparity in the effect of age on the levels of cytokines and of collagen type I and type III in both clinically normal and inflamed gingiva.

The objective of this study was to follow the kinetics of four inflammatory cytokines in the plasma and ascitic fluid of seven patients who developed severe ovarian hyperstimulation syndrome (OHSS) after induction of ovulation for in-vitro fertilization. Blood samples were obtained from these patients at three different times: upon hospitalization; when significant clinical improvement was evident; and after complete resolution. Samples were analysed for interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-8 (IL-8) and tumour necrosis factor alpha (TNF-alpha). Ascitic fluid was obtained by therapeutic paracentesis from all study patients during the active phase and analysed for these cytokines. Two control groups were available: the first included 15 women undergoing controlled ovarian stimulation for in-vitro fertilization without developing OHSS, while the second consisted of 25 healthy women not undergoing ovulation induction or any other medical treatment. High concentrations of IL-1, IL-6 and TNF-alpha were detected in all individuals upon admission for severe OHSS. Concentrations dropped significantly along with clinical improvement, reaching normal values after complete resolution. A statistically significant correlation was found between plasma cytokine concentrations and certain biological characteristics of the syndrome such as leukocytosis, increased haematocrit, and elevated plasma 17-beta-oestradiol concentrations. Ascitic fluid obtained from the study patients contained high IL-6 and IL-8 concentrations, while other cytokines were unaltered. These results suggest close association between inflammatory cytokines and the pathophysiology of the ovarian hyperstimulation syndrome.

OBJECTIVE: To assess serum interleukin-1 beta (IL-1 beta) concentrations in patients with ischaemic heart disease, to characterise subgroups of patients with raised IL-1 beta concentrations, and to examine whether serum IL-1 beta concentrations correlate with non-specific indices of inflammation. DESIGN: Survey study of patients with ischaemic heart disease. SETTING: Cardiac catheterisation laboratory of a tertiary medical centre. PATIENTS: Consecutive patients with angina pectoris and patients recovering from uncomplicated acute myocardial infarction and undergoing elective coronary angiography. RESULTS: Mean(SD) serum IL-1 beta concentrations were higher (P < 0.001) in patients with angina and < 50% coronary artery stenosis (n = 11; 18.8(19.9) pg/ml), patients with angina > or = 50% stenosis (n = 23; 10.2(11.4) pg/ml), and patients 8(0.8) days post-infarction (n = 13; 4.4(5.8) pg/ml) than in 15 healthy, age-matched controls (0.3(0.5) pg/ml). Serum IL-1 beta concentrations did not correlate with total blood leucocyte counts (r = -0.07, P = NS), blood lymphocyte counts (r = -0.24, P = NS), and blood monocyte counts (r = -0.29, P = NS), or with fibrinogen (r = -0.16, P = NS) and C-reactive protein concentrations (9(10.5) mg/dl v 14.1(19) mg/dl for patients with undetectable and detectable concentrations, respectively, P = NS). CONCLUSION: Serum IL-1 beta concentrations are raised in patients with ischaemic heart disease, in particular in those with minimal coronary artery disease and angina. The precise role of IL-1 beta in coronary artery disease remains to be determined.