The Measurement of Intraocular Biomarkers in Various Stages of Proliferative Diabetic Retinopathy Using Multiplex xMAP Technology.

Purpose. To determine the intraocular levels of growth factors and cytokines in patients with various degrees of severity of proliferative diabetic retinopathy (PDR) using multiplex xMAP technology. Methods. A prospective cohort study of 61 eyes from 56 patients who were divided into 3 groups based on the severity of PDR. Patients in group number 1 are those who presented PDR with no need of repeated surgical intervention; patients in group number 2 had repeated vitreous bleeding; and patients in group number 3 had refractory neovascular glaucoma. The concentrations of proangiogenic, antiangiogenic, inflammatory, and neurotrophic factors were measured in intraocular fluid. The results were also compared with levels of factors measured in 50 eyes from 50 patients prior to senile cataract surgery (control group). Results. Patients with refractory neovascular glaucoma (the highest clinical severity group) had higher levels of interleukin 6 (IL-6) (median1 37.19; median3 384.74; P = .00096), transforming growth factor beta 1 (TGFß-1) (median1 49.00; median3 414.40; P = .0017), and vascular endothelial growth factor (VEGF) (median1 211.62; median3 352.82; P = .0454) compared with other PDR patients. Conclusions. Results of our study imply that levels of IL-6, TGFß-1, and VEGF correlate with the severity of PDR.

Differential acute postprandial effects of processed meat and isocaloric vegan meals on the gastrointestinal hormone response in subjects suffering from type 2 diabetes and healthy controls: a randomized crossover study.

BACKGROUND: The intake of meat, particularly processed meat, is a dietary risk factor for diabetes. Meat intake impairs insulin sensitivity and leads to increased oxidative stress. However, its effect on postprandial gastrointestinal hormone (GIH) secretion is unclear. We aimed to investigate the acute effects of two standardized isocaloric meals: a processed hamburger meat meal rich in protein and saturated fat (M-meal) and a vegan meal rich in carbohydrates (V-meal). We hypothesized that the meat meal would lead to abnormal postprandial increases in plasma lipids and oxidative stress markers and impaired GIH responses. METHODS: In a randomized crossover study, 50 patients suffering from type 2 diabetes (T2D) and 50 healthy subjects underwent two 3-h meal tolerance tests. For statistical analyses, repeated-measures ANOVA was performed. RESULTS: The M-meal resulted in a higher postprandial increase in lipids in both groups (p<0.001) and persistent postprandial hyperinsulinemia in patients with diabetes (p<0.001). The plasma glucose levels were significantly higher after the V-meal only at the peak level. The plasma concentrations of glucose-dependent insulinotropic peptide (GIP), peptide tyrosine-tyrosine (PYY) and pancreatic polypeptide (PP) were higher (p<0.05, p<0.001, p<0.001, respectively) and the ghrelin concentration was lower (p<0.001) after the M-meal in healthy subjects. In contrast, the concentrations of GIP, PYY and PP were significantly lower after the M-meal in T2D patients (p<0.001). Compared with the V-meal, the M-meal was associated with a larger increase in lipoperoxidation in T2D patients (p<0.05). CONCLUSION/INTERPRETATION: Our results suggest that the diet composition and the energy content, rather than the carbohydrate count, should be important considerations for dietary management and demonstrate that processed meat consumption is accompanied by impaired GIH responses and increased oxidative stress marker levels in diabetic patients. TRIAL REGISTRATION: ClinicalTrials.gov NCT01572402.

Evaluation of IGF1 serum levels in malignant melanoma and healthy subjects.

BACKGROUND/AIM: There were two aims in the present study. The first was to evaluate the usefulness of insulin-like growth factor 1 (IGF1) for melanoma detection. The second was to correlate changes of serum levels of IGF1 with the Breslow score and sentinel node metastasis positivity. PATIENTS AND METHODS: We examined a group of 216 cases, 77 patients with melanomas and 139 healthy probands. We determined the serum IGF1 levels of each patient using an IRMA radioisotope IGF1 assay kit. Serum samples were collected prior to surgery or any other form of treatment. All melanoma diagnoses were histologically verified. RESULTS AND DISCUSSION: Based on the statistical evaluation between the melanoma group and group of healthy individuals, we observed statistically significant differences in IGF1 serum levels. The median IGF1 levels in the melanoma group was 154.1 ng/ml compared to 111.2 ng/ml in the group of healthy individuals (p=0.0036). The changes of the IGF1 levels related to the Breslow score categories were statistically significant (p=0.0027). Lastly, we compared the results between the positive and negative metastatic affection of the sentinel nodes. The median IGF1 levels in the negative group was 173.5 ng/ml compared to 205.8 ng/ml in the positive group. This difference was statistically significant (p=0.0407). CONCLUSION: Serum levels of IGF1 were significantly higher in patients diagnosed with melanoma compared to the healthy control group. The changes of the IGF1 levels related to the Breslow score categories were statistically significant. Serum levels of IGF1 were significantly higher in the group with the positive metastatic affection of the sentinel nodes than in negative patients.

Postprandial oxidative stress and gastrointestinal hormones: is there a link?

BACKGROUND: Abnormal postprandial elevation of plasma glucose and lipids plays an important role in the pathogenesis of diabetes and strongly predicts cardiovascular mortality. In patients suffering from type 2 diabetes (T2D) postprandial state is associated with oxidative stress, cardiovascular risk and, probably, with impairment of both secretion and the effect of gastrointestinal peptides. Evaluating postprandial changes of gastrointestinal hormones together with changes in oxidative stress markers may help to understand the mechanisms behind the postprandial state in diabetes as well as suggest new preventive and therapeutical strategies. METHODS: A standard meal test has been used for monitoring the postprandial concentrations of gastrointestinal hormones and oxidative stress markers in patients with T2D (n = 50) compared to healthy controls (n = 50). Blood samples were drawn 0, 30, 60, 120 and 180 minutes after the standard meal. RESULTS: Both basal and postprandial plasma concentrations of glucose and insulin proved to be significantly higher in patients with T2D, whereas plasma concentrations of ghrelin showed significantly lower values during the whole meal test. In comparison with healthy controls, both basal and postprandial concentrations of almost all other gastrointestinal hormones and lipoperoxidation were significantly increased while ascorbic acid, reduced glutathione and superoxide dismutase activity were decreased in patients with T2D. A positive relationship was found between changes in GIP and those of glucose and immunoreactive insulin in diabetic patients (p<0.001 and p<0.001, respectively) and between changes in PYY and those of glucose (p<0.01). There was a positive correlation between changes in GIP and PYY and changes in ascorbic acid in patients with T2D (p<0.05 and p<0.001, respectively). CONCLUSION/INTERPRETATION: Apart from a positive relationship of postprandial changes in GIP and PYY with changes in ascorbic acid, there was no direct link observed between gastrointestinal hormones and oxidative stress markers in diabetic patients. TRIAL REGISTRATION: ClinicalTrials.gov NCT01572402.

CA125 and HE4 levels in a Czech female population diagnosed with endometrial cancer in preoperative management.

AIM: The aim of the present study was to compare the use of cancer antigen 125 (CA125) and human epididymis protein 4 (HE4) biomarkers in patients with endometrial cancer for preoperative management and to particularly focus on relationship between CA125 and HE4 and disease stage in predicting myometrial invasion or intrauterine tumor spread. PATIENTS AND METHODS: Thirty-four patients diagnosed with endometrial cancer and 32 healthy controls were enrolled into the pilot study in the period between May 2012 and March 2013. Blood from all the females was collected and examined for CA125 and HE4. Based on standardized ultrasound examination, including gynecological examination, the clinical disease stage was determined. RESULTS: We found a significant difference (p<0.0001) for means of serum levels of HE4: females with endometrial cancer, 75.5 pmol/l, versus healthy females, 40.0 pmol/l. A non-significant statistical difference was found for mean serum CA125 levels (p=0.4442): females with endometrial cancer 19.0 IU/l, versus healthy females, 15 IU/l. A significant correlation with histopathological disease stage was found for both biomarkers (Spearman correlation). Sensitivity and specificity, and the related cut-off for HE4 suggest that HE4 would be a more appropriate biomarker for differential diagnosis between benign and malignant states. CONCLUSION: Based on our pilot study, we found that parallel examination of HE4 and CA125 may support endometrial ultrasound finding verification prior to biopsy. This study is ongoing and we expect that results on a larger population may enable HE4 measurement to be implemented in routine practice.

Defining the seriousness of proliferative vitreoretinopathy by aspiration of cytokines from the anterior chamber.

AIM: Proliferative vitreoretinopathy is the major cause of retinal detachment surgery failure. Our prospective cohort study of 27 eyes aimed to determine intraocular levels of growth factors and cytokines in patients with retinal detachment with various degrees of severity of proliferative vitreoretinopathy using multiplex xMAP(®) Technology. PATIENTS & METHODS: The concentrations of 12 proangiogenic, antiangiogenic, inflammatory and neurotrophic factors were measured from 0.05-ml samples of intraocular fluid using multiplex xMAP Technology. The results were compared with levels of various factors, which were measured in samples from the control group of 31 eyes prior to senile cataract surgery. RESULTS: The concentration of the MCP-1 cytokine was found to be higher in eyes with retinal detachment compared with the control group. The concentration of VEGF was found to be higher in eyes with retinal detachment complicated with proliferative vitreoretinopathy compared with the uncomplicated retinal detachment group and the control group. CONCLUSION: MCP-1 and VEGF may participate in pathogenesis of retinal detachment and proliferative vitreoretinopathy. Biomarkers in disease detection and management have become important tools in modern clinical medicine, and their application to retinal disease should be no exception.

Asymptomatic abdominal aortic aneurysms show histological signs of progression: a quantitative histochemical analysis.

OBJECTIVE: Abdominal aortic aneurysm (AAA) is a serious disease due to its covert nature, relatively high prevalence and fatal prognosis in the case of rupture. To obtain new insights into AAA pathogenesis, we examined the relationships between histopathology, multiplex in vitro immunoassay data, diameter and symptomatology. METHODS: In a prospective, non-randomised study, we evaluated samples from 6 normal infrarenal aortae and 65 AAA patients (65 walls, 55 thrombi). The AAA patients were either asymptomatic (n = 44), symptomatic (n = 7) or with ruptured AAA (n = 14). The AAA diameter was classified as small (<5 cm, n = 18), medium (5-7 cm, n = 26) and large (>7 cm, n = 21). We quantified the histopathology of the AAA wall and the adjacent thrombus. We assessed the expression of proteins in the same samples. RESULTS: Asymptomatic AAAs had walls with more abundant inflammatory infiltrates, lower amounts of PAI-1, a higher number of tPA-positive elements, a tendency towards decreased collagen content, whereas the adjacent thrombi had a greater concentration of VCAM-1 and MMP-2 when compared with symptomatic AAAs. Compared with the aneurysmatic aorta, the normal aorta contained less collagen and more elastin, actin, desmin and PAI-1-positive elements; in addition, it was more vascular. Medium-sized AAAs were the most actin and vimentin rich, and large AAAs were the most vascular. CONCLUSION: Our results show that asymptomatic AAA walls often have more potentially deleterious histopathological alterations than symptomatic AAA walls. This result indicates that a progression from an asymptomatic AAA to rupture can be expected and screening patients who are at risk of rupture could be beneficial.

Plasma TIMP1 level is a prognostic factor in patients with liver metastases.

UNLABELLED: TIMP1 (tissue inhibitor of metalloproteinases 1) regulates extracellular matrix turnover and also promotes cell growth and has anti-apoptotic activity, which promotes malignant processes in tumor tissue. The aim of our study was to evaluate the relation of plasma TIMP1 protein levels with prognosis in patients with liver metastases, with particular regard to possible early-prediction of recurrence of the disease. PATIENTS AND METHODS: We studied a group of 87 patients with metastatic liver disease (mostly from colorectal cancer) who underwent surgery for liver metastases, and assessed their preoperative plasma TIMP1 levels. These levels were evaluated according to prognosis. Furthermore, we measured plasma TIMP1 in the post-operative period and tried to relate the changes with the diagnosis of relapse. RESULTS: We found preoperative plasma TIMP1 levels to be related to overall survival in the group of all patients with metastatic liver disease (p=0.0047), with a higher level being associated with an adverse outcome; the cut-off value was set at 165 ng/ml. This applied to all patients, regardless of the type of surgery. Assessment of the post-operative dynamics of TIMP1 was not found to be statistically significant to indicate disease recurrence. CONCLUSION: We found there to be a relationship between higher plasma levels of TIMP1 and an adverse prognosis in patients with liver metastases. The assessment of plasma TIMP1 levels could help the detection of patients with worse outcome.

HE4 and ROMA index in Czech postmenopausal women.

AIM: The first aim of the project was to evaluate the benefits of the determination of human epididymis protein 4 (HE4) and the risk of ovarian malignancy algorithm (ROMA) index for primary detection of ovarian cancer in a population of Czech women. The second aim was to study the advantages HE4, cancer antigen 125 (CA125) and ROMA index for distinguishing between benign and malignant tumors. Aware of the age distribution of ovarian cancer, we focused on postmenopausal patients. PATIENTS AND METHODS: Our group of patients consisted of 256 females, 21 with ovarian cancer and 235 with benign ovarian tumors. All diagnoses were histologically verified. We determined the serum levels of HE4 and CA125 and calculated the ROMA2 index for postmenopausal women. Serum levels of the analytes were measured using an Architect 1000i instrument. Serum samples were collected prior to surgery or any other form of treatment and the results of the two groups of patients were compared (malignant vs. benign). RESULTS: There was a significant difference in the serum levels for all parameters studied between the groups of patients with malignant and those with benign diagnoses (Wilcoxon test, p<0.0001). When all parameters were evaluated at 95% specificity, the HE4 cut-off was 112 pmol/l at a sensitivity of 71.42%, a positive predictive value (PPV) of 55.56%, a negative predictive value (NPV) of 97.14% and an area under the curve (AUC) of 0.9152. The CA125 cut-off was 81 IU/l at a sensitivity of 80.95%, a PPV of 58.62%, a NPV of 98.23% and an AUC of 0.9731. ROMA2 index had a cut-off 37.70% at a sensitivity of 85.71%, a PPV of 62.06%, a NPV of 98.65% and an AUC of 0.9803. The highest diagnostic efficiency was achieved by the ROMA2 index. CONCLUSION: Determination of HE4 along with CA125 and ROMA2 index calculation is a suitable method for the improvement of the primary detection of ovarian cancer. This approach also improves the differential diagnostic possibilities for distinguishing between malignant and benign tumors.

[Morphology and etiopathogenesis of the abdominal aortic aneurysm]. / Morfologie a etiopatogeneze aneuryzmatu brisní aorty.

The paper summarizes the latest research on the abdominal aorta aneurysm etiopathogenesis and compares normal aorta morphology with changes in the aortic aneurysm wall. The role of risk factors, especially hemodynamic and genetic, is discussed in detail. Special attention is paid to inflammatory processes including cytokines and matrix degrading proteases that contribute to the development of aneurysm. The role of thrombus and the current results of research into biomarkers indicating the risks and progression of the disease are analysed. Finally, a review of pharmacomodulation of the aortic aneurysm using statins, antibiotics, antihypertensive and nonsteroidal antiinflammatory drugs is presented.

Growth factors and breast tumors, comparison of selected growth factors with traditional tumor markers.

BACKGROUND: The first aim of this project was to study new possibilities for distinguishing benign from malignant tumors using growth factors and to compare them with the traditional tumor markers Carcinoembryonic antigen (CEA) and Cancer antigen 15-3 (CA15-3) for breast tumors. The second aim was to make a comparison of CEA, CA 15-3, Insulin-like growth factor I (IGF1), Insulin-like growth factor-binding protein 3 (IGFBP3), Hepatocyte growth factor (HGF) and Epidermal growth factor (EGF) for individual stages of cancer. PATIENTS AND METHODS: Our group of patients consisted of 110 females, 89 with breast cancer and 21 with benign breast tumors (fibroadenomas). Serum levels of CEA and CA 15-3 were measured using a DxI instrument. Serum levels of IGF1 and IGFBP3 were measured using IRMA radioisotope assay kits. HGF and EGF were measured using an xMAP Luminex multiplex panel. Serum samples were collected prior to surgery and those of the two groups of patients were compared (malign vs. benign). Patients with diabetes mellitus were excluded from this project. RESULTS AND DISCUSSION: Comparing the individual parameters of serum levels between the two groups of patients (malignant vs. benign) only HGF was found to show a statistically significant difference. The mean of HGF in patients with malignant diseases prior to surgery was 3370 pg/ml compared to 1799 pg/ml in benign tumors with p=0.0016. We found significantly lower serum values of IGF1 at stage III in comparison to stages I and II: mean values: at stage I=181 ng/ml, at stage II=182 ng/ml and at stage III=70 ng/ml; stage III vs. stage II, p=0.0167. CONCLUSION: Tumor markers are currently used for therapy monitoring in cancer patients as one of the indicators of successful therapy. Our findings correspond to existing literature. IGF1 and its binding protein IGFBP3 cannot be used to distinguish between malignant and benign tumor. HGF is considered to be a marker of progression and of the aggressiveness of breast cancer; our data fully corresponds to this. Based on our data, this marker could potentially be used as an additional tool for the differentiation between benign and malignant tumor.

Prognostic significance of TIMP-1 in non-small cell lung cancer.

UNLABELLED: Tissue inhibitor of metalloproteinases 1 (TIMP1) regulates not only extracellular matrix catabolism but the major effect in tumor tissue is promotion of cell growth and anti-apoptotic activity. The aim of our study was to evaluate plasma TIMP1 levels and tissue TIMP1 mRNA expression as prognostic markers in NSCLC patients. PATIENTS AND METHODS: We studied a group of 108 patients with NSCLC who had undergone lung surgery. Estimation of TIMP1 mRNA was performed by quantitative polymerase chain reaction (qPCR) and estimation of plasma TIMP1 protein using enzyme-linked immunosorbent assay (ELISA). RESULTS: There was shorter disease-free interval (DFI) for NSCLC patients at stage II, with a higher expression of TIMP1 mRNA in tumor tissue (p=0.0246). We recorded a relationship between tumor tissue TIMP1 mRNA expression and DFI in squamous cell carcinoma (SCC) (p=0.0117). Shorter overall survival was found in patients at stages IIIa+IIIb+IV, with a higher expression of TIMP1 mRNA (p=0.0389). We found differences in plasma TIMP1 levels between patients with SCC and those with adenocarcinoma (p=0.0491). CONCLUSION: A higher tissue level of TIMP1 mRNA is related to an adverse prognosis of patients. However, our results did not show any relation of TIMP1 protein in blood plasma to prognosis.

Vitamin D in colorectal, breast, prostate and lung cancer: a pilot study.

BACKGROUND: Many studies have demonstrated the relationship between vitamin D and cancer of many different sites, including of the breast, colorectum, prostate and lung. Most epidemiological studies have assessed the effects of dietary intake only, although endogenous production after sun exposure is the main source of vitamin D. The aim of our pilot study was to study serum levels of vitamin D in general population and in patients with different type of cancer. PATIENTS AND METHODS: The control group consisted of 214 healthy individuals. Pathological groups of patients included 170 patients with different cancer types (28 patients with prostate cancer, 43 patients with breast cancer, 49 patients with colorectal cancer and 50 patients with lung cancer). All of the patients were enrolled in the early clinical stage of cancer up to clinical stage III. Advanced stages were not included into the study. Vitamin D serum levels were measured using ECLIA Roche method. RESULTS: All the results for serum vitamin D from pathological groups were significantly lower compared to the levels of the control group. All the cancer types had a high incidence rate of very low serum levels of vitamin D. Lung cancer had the highest incidence rate of very low vitamin D serum levels. CONCLUSION: We found a high incidence of hypovitaminosis D in cancer patients compared to a healthy control group among a Czech population. This incidence rate is higher in comparison to data found in literature from the other parts of the world. Based on the data from this study, a large epidemiological study monitoring vitamin D serum levels in the healthy population and in cancer patients in the Czech Republic has been already proposed.

The role of MonoTotal in the primary diagnosis, prognosis and follow-up of patients with non-small cell lung cancer (NSCLC).

BACKGROUND: A new cytokeratin tumor marker, MonoTotal was studied in lung cancer, the most common cause of cancer mortality worldwide. In non-small cell lung cancer (NSCLC) patients MonoTotal serum levels and their relationship to the tumor stage, histological subtype, early relapse and cancer-related death were evaluated. PATIENTS AND METHODS: MonoTotal serum levels were studied, using immunoradiometric assay in a group of 93 patients with newly diagnosed NSCLC undergoing radical surgery, and were compared to those with benign lung diseases. RESULTS: A diagnostic power of MonoTotal in distinguishing patients with NSCLC from benign lung diseases was demonstrated. Higher levels of MonoTotal were associated with advanced stages of squamous cell carcinoma and there was a positive correlation of marker with tumor size. Marker levels showed significant relation to disease-free survival and overall survival. CONCLUSION: MonoTotal seems to be a potentially very interesting serum marker that, in conjunction with other clinical data, might be used for monitoring of patients with NSCLC.

Evaluation of tumour markers as differential diagnostic tool in patients with suspicion of liver metastases from breast cancer.

AIM: The liver is the site of breast cancer metastasis in 50% of patients with advanced disease. Tumour markers have been demonstrated as being useful in follow-up of patients with breast cancer, in early detection of recurrence of breast cancer after radical surgical treatments, and in assessing oncologic therapy effect, but no study has been carried out on their usefullness in distinguishing benign liver lesions from breast cancer metastases. The aim of this study was therefore to evaluate the importance of tumour markers carcinoembryonic antigen (CEA), carbohydrate antigen CA19-9 (CA19-9), thymidine kinase (TK), tissue polypeptide antigen (TPA), tissue polypeptide-specific antigen (TPS) and cytokeratin 19 fragment (CYFRA 21-1) in differential diagnosis between benign liver lesions and liver metastases of breast cancer. PATIENTS AND METHODS: The study includes 3 groups: 22 patients with liver metastases of breast cancer; 39 patients with benign liver lesions (hemangioma, focal nodular hyperplasia, liver cyst, hepatocellular adenoma); and 21 patients without any liver disease or lesion that were operated on for benign extrahepatic diseases (groin hernia, varices of lower limbs) as a control group. The serum levels of tumour markers were assessed by means of immunoanalytical methods. RESULTS: Preoperative serum levels of CYFRA 21-1, TPA, TPS and CEA were significantly higher in patients with liver metastases of breast cancer in contrast to healthy controls and patients with benign liver lesions (p-value<0.05). Serum levels of CA19-9 and TK were higher in patients with malignancy in comparison with benign liver disease and healthy controls but these differences were not statistically significant. CONCLUSION: Tumour markers CEA, CYFRA 21-1, TPA and TPS can be recommended as a good tool for differential diagnosis between liver metastases of breast cancer and benign liver lesions.

Predictive value of serum biomarkers in patients after portal vein embolization (PVE): a pilot study.

BACKGROUND/AIM: Insufficient growth of the liver or tumor progression is an important issue of portal vein embolization (PVE) in some patients. This study evaluated the predictive value of serum biomarkers for liver hypertrophy and tumor progression after PVE. PATIENTS AND METHODS: Serum levels of tumor markers, growth factors and cytokines were determined in 40 patients with malignant liver tumors in the pre- and post-PVE period. The values were compared with contralateral liver hypertrophy and tumor progression. RESULTS: Liver tissue hypertrophy occurred in 26 (65%), tumor progression in 11 (27.5%) and insufficient liver hypertrophy in 3 (7.5%) of the patients. The significant predictive biomarkers of PVE included serum TPA levels, monototal, IGF-BP3, IGF1, TGF-α, EGF, HGF, VEGF, TNFa and IL-10 before PVE; and TK, TPA, monototal, IGF-BP3, TGFa and IL-8 over the course of 28 days after PVE. CONCLUSION: Certain serum biomarkers have an important predictive value for the result of PVE.

Changes of cytokine levels during granulocyte-colony-stimulating factor stem cell mobilization in healthy donors: association with mobilization efficiency and potential predictive significance.

BACKGROUND: Peripheral blood stem cells are an important source of hematopoietic stem cells (HSCs) for allogeneic transplantations. Some allogeneic donors mobilize HSCs poorly in response to the granulocyte--colony-stimulating factor (G-CSF). The estimation of the mobilization result in an individual donor is difficult due to the absence of suitable predictive factors. STUDY DESIGN AND METHODS: We analyzed the concentrations and kinetics of certain cytokines induced by G-CSF in 76 healthy donors and compared them with the mobilization efficiency. RESULTS: The levels of the most cytokines increased after the G-CSF application: sICAM, sVCAM, MMP-9, interleukin (IL)-6, TNF-α, sE-selectin, and fibronectin. The concentrations of SDF-1α and IL-8 decreased and VEGF and fractalkine remained unchanged. The premobilization concentrations of IL-6 (p = 0.0093) and TNF-a (p = 0.0006) correlated with preapheresis CD34+ cell count. The comparison of premobilization cytokine levels between better and worse mobilizers showed a difference for TNF-α (p = 0.0006) and IL-6 (p = 0.0682). The TNF-α level below cutoff of 3.6 pg/mL implied approximately 20 times higher risk of poor mobilization (odds ratio, 19.9; p = 0.0002). The immunophenotyping of CD34+ cells suggested a negative correlation between Day +5 CD34+ count and expression of CD11a (p = 0.0319) and a positive correlation with CD44 antigen expression (p = 0.0096). CONCLUSION: The concentrations of certain cytokines corresponded to the quality of HSC mobilization in healthy donors. Their levels measured before mobilization could probably serve as predictive factors for mobilization efficacy and prospectively detect donors who might profit from new mobilization molecules.

Diagnostic and prognostic significance of inflammatory markers in lung cancer-associated pleural effusions.

Besides massive expression in inflammatory pleural effusions, inflammatory markers are also present in cancerinduced pleural effusions. Recent advances in cancer biology point to a role of inflammatory signaling in cancer and encourage reconsidering the diagnostic and prognostic value of inflammatory markers. Here an attempt was made to relate protein levels of inflammatory markers to underlying malignant processes in the pleural space. Pleural effusions from lung cancer patients (n=116) were subjected to a multifactorial analysis covering 13 inflammatory markers. The composition of tumor-associated effusions was compared with that of parainflammatory pleural effusions (n=30), transudates (n=18), and serum values, and evaluated in relation to cancer origin, histology, cytology, pleural involvement, treatment history, and survival time. Inflammatory markers were significantly expressed in pleural effusions of paraneoplastic origin when compared to transudates and most serum levels. Values in pleura-invading and metastatic tumor-associated effusions were typically higher than those of other tumors. Many markers correlated negatively with survival, most prominently IL-8 (r=-0.36, p=0.001) and VEGF (r=-0.35, p=0.001). It appears that most inflammatory markers are highly expressed in tumor-associated pleural effusions, reflecting to some extent tumor origin and localization. Despite the lower efficacy of inflammatory markers in the differentiation between exudative pleural effusions, some inflammatory markers may represent potential prognostic markers of malignant processes in the pleural space.

Effect of hyperinsulinemia and very-low-calorie diet on interstitial cytokine levels in subcutaneous adipose tissue of obese women.

Type 2 diabetes and obesity are associated with an enhanced release of a number of adipocytokines. Hyperinsulinemia, frequently present in type 2 diabetes and obesity, might be one of the drivers of the enhanced production of adipocytokines. The aim of this study was to investigate the interstitial levels of cytokines in subcutaneous adipose tissue (SCAT) in response to hyperinsulinemia and the effect of weight-reducing hypocaloric diet on this regulation in obese subjects. Thirteen obese premenopausal women participated in the study. Concentrations of seven cytokines were measured in plasma and in AT interstitial fluid collected by microdialysis during a euglycemic-hyperinsulinemic clamp and during control infusion of physiological saline. A subgroup of six women underwent a 4-wk very-low-calorie diet (VLCD). Microdialysis during the clamp was performed before and at the end of VLCD. Hyperinsulinemia induced an increase of monocyte chemoatractant protein (MCP-1) and IL-6 SCAT interstitial and plasma levels and elevated IL-8 levels in SCAT. The relative changes of IL-6 levels in the dialysate correlated with changes of IL-8 and MCP-1. The interstitial and plasma levels of IL-1ß, IL-10, TNFα, and plasminogen activator inhibitor (PAI-1) remained unchanged in response to hyperinsulinemia. VLCD resulted in enhancement of the hyperinsulinemia-induced augmentation of MCP-1, IL-6, and IL-8 interstitial levels. In conclusion, hyperinsulinemia upregulates the interstitial levels of MCP-1, IL-6, and IL-8 in SCAT in obese women, whereas it does not affect IL-1ß, IL-10, TNFα, and PAI-1 levels. Hypocaloric diet associated with weight reduction enhances the hyperinsulinemia-induced upregulation of MCP-1, IL-6, and IL-8 in SCAT.

Test of ovarian cancer multiplex xMAP technology panel.

BACKGROUND: An ovarian cancer marker panel including leptin, prolactin, osteopontin, insulin-like growth factor II (IGFII), macrophage migration inhibitory factor and CA125 was tested for multiplex marker measurement. Multiplex was compared to single assayed tumour markers: CA125, TPS, thymidine kinase, monototal and HE4. PATIENTS AND METHODS: The serum marker levels in ovarian cancer patients were compared to controls with benign ovarian disease. xMAP technology was used for multiplex panel and routine immunoanalytic methods for other markers. RESULTS: Considering the multiplexed markers, only CA125(Luminex), osteopontin and IGF-II differed significantly between groups. Levels of all non-multiplexed tumour markers were significantly higher in the cancer group compare to the control. CONCLUSION: Multiplex is a powerful tool for multiparametric studies, but we are still in the era of looking for the right marker combinations for cancer diagnostics and monitoring. The panel will be tested on a larger ovarian cancer cohort and in patients with other cancer and non-cancerous diseases.