Multiplex cytokine detection versus ELISA for aqueous humor: IL-5, IL-10, and IFNgamma profiles in uveitis

PURPOSE: The purpose of this study was to determine levels of IL-2, -4, -5, -10, TNF-alpha, and IFN-gamma in aqueous humor (AH) from patients with active panuveitis, anterior uveitis (AU), and noninflammatory controls by using a flow cytometric mutiplex array (CBA) and to compare with results from ELISA. METHODS: Pooled normal AH was spiked with six cytokines at decreasing concentrations for evaluating the CBA. AH was also obtained from 10 controls (cataract patients) and 36 patients with active uveitis. Cell-free supernatants were added to a cocktail of capture beads and detector antibodies or to antibody-coated wells for CBA and ELISA determination, respectively. RESULTS: CBA demonstrated greater sensitivity for detecting IL-4, IL-10, and TNF-alpha than with ELISA. Increased IFN-gamma was detected in both AU and panuveitis groups compared with controls (P < 0.01). IL-10 was higher in the panuveitis group on steroids (P < 0.01). IL-5 was detected in the control (P < 0.01) and AU groups (P < 0.05) but was undetectable in the panuveitis group (n = 10). Correlations between IFN-gamma and IL-10 were found in all uveitis groups (P < 0.01) but not in controls, whereas TNF-alpha correlations with IL-4/IFN-gamma were obtained in controls but not in the uveitis groups (P < 0.01). CONCLUSIONS: It was possible to measure cytokines titrated into normal AH specimens by CBA, and a greater number of cytokines were detected with increased sensitivity than with ELISA. Elevated IFN-gamma in active uveitis and decreased IL-5 in posterior uveitis suggest Th1 polarity is more marked, with greater uveal tract involvement. The increased IL-10 in the steroid treated group suggests glucocorticoid-induced IL-10 upregulation.     

Non-typical leukemic metastases localization in bulbus oculi and vertebral column and in thyroid gland and trachea in the course of chronic lymphocytic leukemia--two case reports

Chronic lymphocytic leukemia (CLL) is characterized by the clonal proliferation of small mature lymphocytes with their accumulation in blood, bone marrow, lymph nodes, spleen and lien. In advanced stages of CLL, the leukemic metastases can occur in skin, pleura and lungs. They are often strictly connected with disease progression and are present with advanced lymphadenopathy and hepatosplenomegaly, high leukocytosis 100G/1 with 80-90% lymphocytes in blood smear, anemia Hb <10 g% and thrombocytopenia PLT < 100 G/L. In this article we present two CLL patients with non typical leukemic metastases localization in bulbus oculi, vertebral column (case 1) and in thyroid gland and trachea (case 2), that occurred in hematological stabilization phase of disease during normal lymphocytes number or a little lymphocytosis in peripheral blood and a lack of progression in lymphoid organs.     

A long term survival of the patient treated due to advanced adrenocortical carcinoma

The authors presented a case of 52-years old woman with advanced adrenocortical carcinoma completely recovered after surgical resection followed by chemotherapy and mitotane treatment. Two years later metastatic tumor of hepar was excised. From the first diagnosis - patient lives 12 years, without any symptoms of recurrence until now. The authors accentuate a big value of postoperative control, especially imaging studies for early diagnosis of recurrent of cancer and beginning therapy.     

TT virus infection and pancreatic cancer： Relationship or accidental coexistence

TT virus (TTV) was first isolated in 1997 from the patient with acute post-transfusion hepatitis. This fact led to the conclusion that the virus was hepatotropic and could be one of the causative agents of acute hepatitis. Afterwards, however, the virus was found in other human tissues and serological studies revealed that it was widespread. Multiple tropisms of TTV and the fact of its high incidence in general population are considered to indicate no medical significance of TTV in human pathology. Here we present a report of two cases of TTV infection in patients who developed pancreas cancer. The patients were hospitalized at the Department of Infectious Diseases due to hepatitis of unknown origin. Since serological and virological markers of common primary and secondary hepatotropic viruses were negative, TTV-DNA was found in serum and was believed to be the only causative agent with probable hepatotropic action. The patients later developed pancreas cancer and they underwent operation. The relationship is difficult to confirm, however the cases we present should be treated as a preliminary report and a comment on the real role of TTV in human pathology.     

Molecular analysis of uropathogenic strains of Escherichia coli and the clinical course of urinary tract infection in children

The most frequent etiological factor of urinary tract infections are virulent Escherichia coli strains. Identification of uropathogenic Escherichia coli strains is possible using biomolecular techniques. THE PURPOSE OF THIS STUDY was to determine the prevalence of Escherichia coli strains encoding papG adhesins and adhesins of Dr family in children with urinary tract infections and to establish the relationship between the bacterial genotype and clinical course of a sickness. MATERIAL AND METHODS: In 163 children with urinary tract infection caused by Escherichia coli strains, after taking history and physical examination, inflammatory parameters were determined. In all patients abdominal ultrasonography and in chosen patients -- miction cystourethrography and urodynamic examinations were performed. In order to identify Escherichia coli strains possessing pap gene cluster class I, II and III (encoding P fimbriae) and genes encoding Dr family of adhesins, the Polymerase Chain Reaction was used. RESULTS AND CONCLUSIONS: From all isolated Escherichia coli strains, pap gene cluster was identified in 41.1% and dra gene cluster in 42.3%. Escherichia coli strains encoding papG gene class II frequently caused upper urinary tract injections, while strains encoding papG gene class III and genes encoding adhesins of Dr family were mostly responsible for lower urinary tract infections and their recurrences. Occurrence of virulent Escherichia coli strains was not more common in children with pyelonephritis and with disorders of urinary tract in comparison with the group of children without urinary tract anomalies. Determination of bacterial virulence factors causing urinary tract infections in children may be helpful in the prognosis of the course of the disease.     

Prediction of the excessive perioperative bleeding in patients undergoing coronary artery bypass grafting: role of aspirin and platelet glycoprotein IIIa polymorphism

OBJECTIVE: The presence of the glycoprotein IIIa allele PlA2 is associated with enhanced thrombin formation and an impaired antithrombotic action of aspirin, which could favor coronary thrombosis. We wondered whether PlA1/A2 genetic polymorphism could affect the postoperative bleeding in patients undergoing coronary artery bypass grafting. We also aimed to assess the effects of aspirin pretreatment and to ascertain the value of platelet function studies as predictors of postoperative bleeding. METHODS: In a randomized, double-blind study, patients undergoing coronary artery bypass grafting were pretreated with a 150-mg dose of aspirin orally 12 and 3 hours before surgery (n = 51, 41 elective) or with placebo (n = 51, 43 elective). The hemostasis was monitored by Simplate (bioMérieux, Inc, Durham, NC) bleeding time and capillary closure time (platelet function analyzer PFA 100; Sysmex UK Ltd, Milton Keynes, United Kingdom). Postoperative bleeding and blood products transfusions were recorded. The glycoprotein IIIa polymorphism was analyzed. RESULTS: Bleeding was significantly greater in PlA1 homozygotes from control group. Blood loss was significantly greater (by 25%) in aspirin group. The volume of blood products transfusions in aspirin patients was significantly larger (by 137%). When subjects were stratified accordingly to blood platelet glycoprotein IIb/IIIa genotype, in the aspirin group PlA2 carriers had greater blood loss than PlA1 homozygotes (1858 +/- 932 mL vs 1216 +/- 525 mL, P < .05). CONCLUSION: PlA1 homozygotes normally had a greater risk of perioperative bleeding. Capillary closure time had no advantage relative to Simplate bleeding time in predicting postoperative blood loss. Aspirin pretreatment revealed no beneficial effects and resulted in increased postoperative bleeding and requirement for blood product transfusions after coronary artery bypass grafting in patients with stable angina. It was most unfavorable for PlA2 carriers.     

Metabolic and hormonal profiles: HDL cholesterol as a plausible biomarker of breast cancer risk. The Norwegian EBBA Study.

Low serum high-density lipoprotein cholesterol (HDL-C) is an important component of the metabolic syndrome and has recently been related to increased breast cancer risk in overweight and obese women. We therefore questioned whether serum HDL-C might be a biologically sound marker of breast cancer risk. We obtained cross-sectional data among 206 healthy women ages 25 to 35 years who participated in the Norwegian EBBA study. We included salivary ovarian steroid concentrations assessed by daily samples throughout one entire menstrual cycle, metabolic profile with measures of adiposity [body mass index (BMI) and truncal fat percentage], serum concentrations of lipids and hormones (insulin, leptin, testosterone, dehydroepiandrostendione sulfate, insulin-like growth factor-I, and its principal binding protein), and mammographic parenchymal pattern. We examined how components of the metabolic syndrome, including low serum HDL-C, were related to levels of hormones, and free estradiol concentration in particular, and studied predictors of mammographic parenchymal patterns in regression models. In women with BMI > or = 23.6 kg/m(2) (median), overall average salivary estradiol concentration dropped by 2.4 pmol/L (0.7 pg/mL; 13.2% change in mean for the total population) by each 0.33 mmol/L (12.8 mg/dl; 1SD) increase in serum HDL-C (P = 0.03; P(interaction) = 0.03). A subgroup of women characterized by both relatively high BMI (> or =23.6 kg/m(2)) and high serum LDL-C/HDL-C ratio (> or = 2.08; 75 percentile) had substantially higher levels of salivary estradiol by cycle day than other women (P = 0.001). BMI was the strongest predictor of overall average estradiol with a direct relationship (P< 0.001). Serum HDL-C was inversely related to serum leptin, insulin, and dehydroepiandrostendione sulfate (P < 0.001, P < 0.01, and P < 0.05, respectively). There was a direct relationship between breast density and healthy metabolic profiles (low BMI, high serum HDL-C; P < 0.001) and salivary progesterone concentrations (P < 0.05). Our findings support the hypothesis that low serum HDL-C might reflect an unfavorable hormonal profile with, in particular, increased levels of estrogens and gives further clues to biomarkers of breast cancer risk especially in overweight and obese women.     

Circulating D-dimer levels are better predictors of overall survival and disease progression than carcinoembryonic antigen levels in patients with metastatic colorectal carcinoma

BACKGROUND: Fibrin formation is required for tumor angiogenesis, metastasis, and invasion. D-dimer, a fibrin degradation product, is produced when crosslinked fibrin is degraded by plasmin. The current study prospectively examined D-dimer levels in patients with metastatic colorectal carcinoma treated in a Phase II randomized trial comparing bevacizumab (Avastin, Genentech, South San Francisco, CA) plus 5-fluorouracil/leucovorin (5-FU/LV) with 5-FU/LV alone. METHODS: At least one circulating D-dimer level was evaluable in 98 of the 104 previously untreated patients with metastatic colorectal carcinoma in the current trial. Plasma D-dimer levels were determined using a quantitative immunoassay kit at enrollment, before each treatment, and at the time of trial completion or disease progression. RESULTS: At trial enrollment, 86 of 104 patients (88%) had elevated D-dimer levels (> 20 ng/mL), and 86 of 102 patients (84%) had elevated carcinoembryonic antigen (CEA) levels (> 3 ng/mL). Baseline D-dimer levels were correlated with the following baseline characteristics: CEA (Pearson coefficient, 0.31; P = 0.002), albumin levels (Pearson coefficient, -0.32; P = 0.002), tumor burden (Pearson coefficient, 0.30; P = 0.003), and number of metastatic sites (Pearson coefficient, 0.21; P = 0.04). At the time of progression, plasma D-dimer levels reached a maximum postbaseline value in 51 of 61 patients (84%), whereas the CEA level was at its maximum postbaseline value in 39 of 55 patients (71%). Baseline D-dimer levels were a strong predictor of overall survival on univariate analysis (P = 0.008) and multivariate analysis (P = 0.03). Overall, treatment with bevacizumab (5 mg/kg) and baseline D-dimer levels were the only predictors of overall survival (P < 0.05). CONCLUSIONS: The current study indicates that fibrin remodeling is an important prognostic feature in metastatic colorectal carcinoma. D-dimer levels should be incorporated into prognostic models, and D-dimer may represent a useful biomarker for patients treated with antiangiogenic agents.