Band 49, Heft 10

Circulating tumor cells (CTCs) provide unique information for the management of breast cancer patients, since their detection and monitoring is useful for prognosis, prediction of response to therapy, or monitoring clinical course in patients with localized or metastatic disease. Currently, the most practical application of CTCs is monitoring of patients with metastatic disease. Elevated CTC levels prior to initiation of a new systemic therapy are associated with a worse prognosis while persistently elevated CTC levels strongly suggest that the therapeutic regimen with which the patient is being treated is not working. New areas of research are directed toward developing novel sensitive assays for CTC molecular characterization. Molecular characterization of CTCs is very important for the future use of CTCs as targets of novel therapies. This review has focused on the presentation of recent data showing that CTCs are emerging as novel tumor biomarkers for prognostic and predictive purposes in breast cancer.

Non-small cell lung carcinomas (NSCLC) account for about 80% of lung cancers and their remarkable heterogeneity manifests in histology, pathogenesis, prognosis, and response to treatments. Recent advances in molecular characterization help stratifying NSCLC patients for their potential benefit from targeting therapies. However, the fundamental mechanisms underlying the tumoral heterogeneity remain poorly understood. Expression profiling of many microRNAs (miRNAs) in various normal and disease tissues demonstrated unique spatial and temporal expression patterns and some miRNAs have been functionally characterized as oncogenes or tumor suppressor genes. Genome-wide screening identified specific miRNA expression signatures associated with clinical outcome of NSCLC patients. A group of miRNAs that has enriched expression in normal lung was found down regulated in NSCLC and may function as tumor suppressor genes. In this review we: a) summarize the current understanding of the critical role that miRNAs play in normal cell functions and in disease biology especially in lung cancer tumorigenesis, b) highlight their potential as biomarkers for lung cancer risk stratification, outcome prediction and classification of histologic subtypes, c) critically assess current knowledge on lung-enriched miRNAs and expression of their predicted target genes in NSCLC and d) evaluate their potential as circulating biomarkers and therapeutic targets in lung cancer.

Tumour markers are a very heterogeneous group of molecules that are generally found in very small concentrations in the plasma and serum of healthy individuals. In the process of neoplastic differentiation the cell can synthesize, release, or induce synthesis of other cells, thus increasing their concentration in plasma and serum. These substances may also increase their plasma concentration in patients without cancer due to processes that increase the release or reduce catabolism, and so give rise to false positives. An understanding of the main physiopathological processes that increase the concentrations of these substances could improve our interpretation of tumour markers and their clinical application. In this study we review the physiopathological processes that may increase the plasma concentrations of tumour markers. We performed a bibliography review in PubMed, searching for causes of false positives for the following tumour markers: α-Fetoprotein, CA 125, CA 15-3, CA 19-9, CA 72-4, carcinoembryonic antigen, CYFRA 21-1, squamous cell carcinoma, prostatic specific antigen, β 2 -microglobulin, choriogonadotropin (β chain), chromogranin A, neuron specific enolase, HER2-neu, progastrin releas-ing peptide, S-100, and thyroglobulin. The results favour the use of tests which can identify pathological processes that may increase tumour marker concentrations.

Recent advances have provided unprecedented opportunities to identify prognostic and predictive markers of efficacy of cancer therapy. Genetic markers can be used to exclude patients who will not benefit from therapy, exclude patients at high risk of severe toxicity and adjust dosing. Genomic approaches for marker discovery now include genome-wide association studies and tumor DNA sequencing. The challenge is now to select markers for which there is enough evidence to transition them to the clinic. The hurdles include the inherent low frequency of many of these markers, the lengthy validation process through trials, as well as legislative and economic hurdles. Attempts to answer questions about certain markers more quickly have led to an increased popularity of trials with enrichment design, especially in light of the dramatic phase I results seen in recent months. Personalized medicine in oncology is a step closer to reality.

Background: The genotyping of HFE p.C282Y and p.H63D mutations is one of the most requested molecular analyses in the laboratorial routine. In this scenario, the main aim was to develop a genotyping assay that has advantages compared to other methods. Methods: Genotypes for the HFE p.C282Y (c.G845A; rs1800562) and p.H63D (c.C187G, rs1799945) mutations were assessed by polymerase chain reaction (PCR) followed by high resolution melting (HRM) analysis with the Rotor-Gene 6000 ® instrument. Validation studies were conducted in samples bi-directionally sequenced. Results: The melting assay was developed in a unique procedure and to ensure the result in approximately 112 min (31 min for sample preparation and 81 min for the PCR-HRM step). Genotypes for the HFE p.C282Y mutation were easily distinguished in the region of 80–86°C. For the HFE p.H63D, genotypes were also easily distinguished in the region of 76–82°C, but using the addition of known wild-type genotype DNA in all unknown samples plus a reaction without addition. In validation, genotypes were 100% concordant between methods. Conclusions: Our genotyping assay with the Rotor-Gene 6000 ® instrument applies to the laboratorial routine with several advantages, especially in large-scale demand. The main advantages were the non-dependence on gel electrophoresis and on mutagenic reagents for visualization of fragments, reduction of the chances for contamination due to sample preparation, the lack of use of probe-based methods and cost-effectiveness.

Background: The goal is to introduce visual performance mapping efficient for establishing acceptance criteria and facilitating decisions regarding the utility of hospital point-of-care devices. This approach uniquely reveals the quality of performance locally , as opposed to globally. Methods: After presenting theoretical foundations, this study illustrates the approach by applying it to six hospital glucose meter systems (GMSs) using clinical multi-center (n=2767) and multi-system (n=613, n=100) observations. Results: LS MAD curves identified breakouts, that is, points where the locally-smoothed median absolute difference (LS MAD) curve exceeds the recommended error tolerance limit of 5 mg/dL (0.28 mmol/L). LS maximum absolute difference (MaxAD) breakthroughs, which occur where the LS MaxAD curve exceeds the 99th percentile of MaxADs from x=30–200 mg/dL (1.67–11.10 mmol/L), showed extreme error locations. A multi-sensor interference- and hematocrirt-correcting GMS displayed a flat LS MAD curve until it reached a breakout of 179 mg/dL (9.94 mmol/L) and generated breakthroughs that could affect bedside decision-making, but less erratically than other systems with inadequate performance for hospital critical care. We discovered Class I (meter high, reference low) and Class II (converse) discrepant values in some systems. Class I errors could lead to inappropriate insulin dosing and hypoglycemic episodes in tight glucose control. Conclusions: LS MAD-MaxAD curves help assess the performance of point-of-care testing. Visual mapping of systematic and random errors locally over the entire analyte measurement range in a single integrated display is an advantage when considering the adverse impact of zones of poor quantitative performance on specific clinical applications, threshold-driven bedside decisions and the care of critically ill patients.

Background: Complications after orthopaedic surgery have a substantial impact on resources, including the financial resources of the health-care system. Early detection of patients developing complications, thus excluding those patients without complications should have a major impact on patients’ management. Methods: We evaluated the prognostic value of blood levels of inflammatory markers (interleukin-6, C-reactive protein, total white blood cell count) for complications after primary hip and knee arthroplasty in a routine setting, and their impact on patient management. First, the three inflammatory markers were evaluated in 68 consecutive patients treated at the Department of Orthopaedics. Laboratory markers were measured on days 2 and 4 after surgery. Second, patient management before and after adapting the routine order for laboratory parameters after arthroplasty were evaluated. Results: We could show that interleukin-6 and the C-reactive protein had an almost equal negative predictive value for the exclusion of postsurgical complications. Nevertheless, interleukin-6 returned below its reference levels almost 2 days earlier compared to the other markers. This made an earlier discharge of patients with presumable complications-less postsurgical process possible. Conclusions: So, the total number of patients treated as well as the financial reimbursement could be increased and concurrently occupancy days and averaged length of stay were reduced.

Background: Pentraxin 3 (PTX3) plays an important role in innate immune responses and in inflammation disease. The aim of this study was to investigate the diagnostic and prognostic potential of PTX3 in pelvic inflammatory disease (PID) and correlate it with the severity and outcome of PID. Methods: Blood specimens were collected from 64 patients with PID before and after treatment and 70 healthy controls and the plasma levels of PTX3 were measured using enzyme-linked immunosorbent assay (ELISA) kits. Results: It was found that the plasma level of PTX3 expression was elevated in PID patients compared with healthy controls and decreased significantly after they received treatment. When the cut-off level of plasma PTX3 was set at 2.87 ng/mL, PTX3 had higher sensitivity (84.38%) and lower false-negative rate (15.63%) than CRP (79.69% and 20.31%, respectively) in predicting PID. The level of PTX3 also exhibited a significant correlation with length of hospital stay (r=0.581, p<0.001). Conclusions: Plasma PTX3 concentration not only predicts the presence of PID with lower false-negative rate than CRP, but plasma PTX3 concentration is also affiliated with the presence of tubo-ovarian abscess (TOA) and the length of hospital stay.

Background: The association between coffee intake and circulating levels of C-reactive protein (CRP) may be modified by oxidative stress. The authors examined the relation of coffee consumption to serum CRP considering potential inter-actions of serum γ-glutamyltransferase (GGT) and bilirubin. Methods: The subjects included 4455 men and 5942 women aged 49–76 years who participated in the baseline survey of a cohort study on lifestyle-related diseases in Fukuoka, Japan. Geometric means of serum CRP and 95% confidence intervals across the category of coffee intake stratified by serum GGT and bilirubin were estimated using multiple linear regression. Results: Serum CRP concentrations were progressively lower with higher intake of coffee in men with high serum GGT (p for trend=0.009), but not in those with low serum GGT (p for trend=0.73) and GGT modified the association (p for interaction=0.03). Women showed no association between coffee intake and CRP whether serum GGT was low or high. There was no effect modification of serum bilirubin on the association between coffee intake and CRP in either men or women. Conclusions: These results support a protective effect of coffee intake against systematic inflammation in middle-aged and elderly Japanese men and imply that such an effect may be stronger in elevated oxidative stress.

Background: In order to evaluate whether there is still present a very large variability in measured values and analytical performance among different free thyroid (FT) hormone immunoassays, we considered the results derived from an External Quality Assessment (EQA) scheme, named Immunocheck. Methods: The EQA Immonocheck study enrolled about 1000 participant laboratories, which measured the 54 quality control samples distributed in the three annual cycles (2007, 2008 and 2009). Participant laboratories produced a total of 30,476 results for FT3 and 31,351 for FT4, respectively. Results: The results recovered during the EQA cycles allowed the estimation of assay imprecision (i.e., within-method and between-laboratories variability). On average, control samples with lower free triiodothyronine (FT3) and free thyroxine (FT4) concentrations showed higher imprecision values (CV%) than those with levels within or above the normal range. The agreement among the results produced by different methods was estimated by computing the percent differences (i.e., percent bias values) between the concentrations measured by the methods and the mean of all collected results (i.e., consensus mean). Conclusions: The results of our study demonstrate that a large variability in measured values is still present among different free thyroid hormone immunoassays. Indeed, some immunoassays for both FT3 and FT4 measurement showed percent bias values compared to the consensus mean >20%. Laboratories should inform the clinicians about analytical performance and reference limits of the method used. Furthermore, the clinicians should avoid the use of different methods in the follow-up of patients.

Background: Transferrin (Tf) glycoform lacking one or two complete or incomplete glycan chains (i.e., asialo-monosialo- and disialo-Tf) typically appear in blood after chronic alcohol consumption, though recently it was reported that monosialo-Tf is associated with trisialo-Tf but not with alcohol consumption. These glycoforms are collectively known as carbohydrate-deficient transferrin (CDT). Since samples from alcoholic patients are characterized by decreased sialic acid content in serum transferrin, the assessment of CDT is thereby widely used for laboratory evaluation of chronic alcohol abuse. Methods: CDT analysis has been performed in 6011 consecutive subjects undergoing national mandatory testing after the confiscation of driving license for driving under the influence of alcohol. Out of the 6011 specimens, 539 (9%) displayed values exceeding the specific cut-off (>1.3%) on multicapillary electrophoresis (MCE) (Capillarys2 Sebia, France), and were further analyzed with a routine high-pressure liquid chromatography (HPLC) technique. Results: The overall correlation between the methods in the total 539 samples was satisfactory, displaying a correlation coefficient (r) of 0.960. Nevertheless, the correlation was lower in the group with CDT values comprised between 1.3% and 1.9% (group 1; r=0.60) than in those with CDT values >2.0% (group 2; r=0.98). Moreover, the discordance between values exceeding the method-specific threshold in the former group of samples was also remarkably high (62% of samples in group 1 vs. 0.6% in group 2). Finally, a significant difference of CDT values was observed in group 1 (p<0.001), and in group 2 (p<0.0001) by Wilcoxon test. Conclusions: The MCE is characterized by a high throughput and it seems a suitable approach for laboratory monitoring of alcohol abuse when CDT is used as medical parameter in the diagnosis and follow-up of heavy drinking. However, CDT measured by screening techniques must be confirmed by a confirmatory technique, in particular for forensic purpose.

Background: Acute coronary syndromes present clinically as a consequence of plaque rupture and thrombosis possibly related to altered homeostasis of thrombogenic factors. It is speculated that this vulnerability in adults should be predictable from blood levels of thrombogenic biomarkers in children and adolescents. This study aims to examine the determinants and blood levels of lipoprotein(a) [Lp(a)], fibrinogen (FBG) and plasminogen activator inhibitor-1 (PAI-1) in healthy adolescents stratified according to age group, gender and body mass. Methods: A total of 774 (316 males 458 females) healthy adolescent Arab subjects aged 10–19 years and attending secondary schools in Kuwait were interviewed by a validated questionnaire for variables relating to socio-demographic variables, diet and physical activity. They also had anthropometry, BP measurement and determination of fasting blood levels of Lp(a), low density lipoprotein (LDL)-cholesterol, apolipoprotein (apo) B, PAI-1 activity and FBG. Results: The median (interquartile range, IQR) plasma levels of PAI-1 activity, FBG, Lp(a) and apoB were respectively 1.59 (0.58–3.78) U/mL, 296 (190–417) mg/dL, 10.0 (4.8–21.0) mg/dL and 0.72 (0.60–0.85) g/L. Boys had significantly higher PAI-1, FBG and apoB concentrations than the girls, although Lp(a) levels were greater in the latter. The overweight and obese subjects tended to have higher levels of LDL, apoB, FBG and PAI-1 but not Lp(a). Furthermore, the younger adolescent males and females (age <14 years) consistently had higher FBG levels than the older ones (age >14 years). Lp(a) and PAI-1 levels did not appear significantly influenced by this age stratification. Bivariate and multivariate analyses with adjustment for putative body mass index (BMI) confounders indicated that the independent determinants of these biomarkers were (i) Lp(a): apoB, gender; (ii) PAI-1: BMI, apoB, diet; (iii) FBG: BMI, gender, age, family income; and (iv) apoB: BMI, gender and PAI-1. Conclusions: The blood levels of the prothrombotic biomarkers ;ibLp(a), PAI-1, and FBG;ic in healthy Kuwaiti adolescent subjects are variably influenced by age, gender, body mass and socio-demographic factors.

Background: Multiplexed cytokine measurement offers many advantages over the conventional enzyme-linked immunosorbent assay (ELISA) format when applied in large- scale epidemiological studies or clinical trials. In the present study we set out to define the reliability and consistency of a suspension multiplexed protein array, the cytometric bead array (CBA), in large-scale, longitudinal studies. Methods: The cytokines interleukin (IL)-5, IL-10, tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) were measured in pediatric samples from childhood asthma and allergy studies. Analytical performance of CBA was determined in sample supernatants and CBA was compared to conventional ELISA. Results: Within-run and total imprecision were between 5.2%–10.8% and 5.6%–13.2%, respectively, at three different concentrations for all cytokines. Slopes of dilution linearity were between 1.01 and 1.31 for the four cytokines. The recovery rate at two different concentrations of the cytokines was between 97% and 113%. Lower limits of detection and quantification as well as functional sensitivity were determined. Comparison of the multiplex array and solid phase method showed good correlation with r between 0.82 and 0.93. The sample volume required for the multiplex format was 25% of the ELISA sample volume. Conclusions: CBA analytical evaluation and comparison to an ELISA format demonstrated high reproducibility, sensitivity and good applicability for small volume samples.

Background: Gastrointestinal/liver graft-vs.-host disease is a frequent complication after allogeneic hematopoietic cell transplantation. Endoscopic biopsies are needed to confirm clinical diagnosis, but this is not always feasible due to concurrent complications after transplantation. Cytokeratin 18 is expressed in epithelial colon cells and hepatocytes. Apoptosis, a hallmark of graft-vs.-host disease, results in the cleavage of cytokeratin 18 and the resulting fragments are released into the circulation. Methods: The aim of the present study was to assess the general performance and usefulness of a serologic test for soluble cytokeratin 18 for gastrointestinal/liver graft-vs.-host disease detection. Plasmatic concentration of soluble cytokeratin 18 was measured in 38 individuals undergoing hematopoietic cell transplantation. A two-fold increase of sCK18 above the pre-transplant concentration was used as a threshold value. Results: Plasmatic concentration of soluble cytokeratin 18 was significantly elevated in patients with gastrointestinal/liver graft-vs.-host disease. Soluble cytokeratin 18 concentration raised before the onset of clinical symptoms in 69% of the patients with gastrointestinal/liver graft-vs.-host disease. The threshold value used in our study resulted in a high sensitivity and a low false negative rate for gastrointestinal/liver graft-vs.-host disease detection. Conclusions: Soluble cytokeratin 18 seems to be a suitable biomarker for gastrointestinal/liver graft-vs.-host disease detection, particularly when a biopsy is not feasible.

Background: The objective of the present study is to explore the association between zinc concentrations and insulin-like growth factor 1 (IGF-1), its binding protein (IGFBP-3) and total prostate-specific antigen (tPSA) levels in the serum of patients with prostate cancer (PCa) and prostate intraepithelial neoplasia (PIN), a pre-cancer state matched for age. Methods: The study was carried out in a group of 229 patients who had transurethral prostate biopsy performed. The patients were divided into three groups: control group (BPH), PIN group or PCa group. The patients had plasma zinc concentration determined by atomic absorption spectrometry; IGF-1, IGFBP-3 analyzed using the chemiluminescence method and tPSA detected in serum with DELFIA assay. Results: The studies revealed that, in PCa and PIN patients aged under 65 years, mean zinc concentrations were significantly lower compared with the control group. IGF-1 level significantly increased with decreasing level of zinc in plasma, hence a significantly decreased Zn/IGF-1 ratio. The mean tPSA concentration was significantly increased only in PCa patients of both age groups, whereas the Zn/tPSA ratio significantly decreased with increasing severity of neoplastic lesions, particularly in patients aged under 65 years. Statistical significance was noted for IGF-1:tPSA and IGFBP-3:tPSA ratios, being almost two-fold lower in the PCa patients than in the control group. Conclusions: A significantly lowered Zn/tPSA ratio appears to be a sensitive marker of neoplastic lesions, PCa and PIN, regardless of age. In men under 65 years, the Zn/IGF-1 ratio was reduced, depending on the stage of neoplastic lesions (PIN>PCa). These finding can be useful in early diagnosis of prostate cancer.

Background: CDC25 phosphatases control cell cycle progression by activating cyclin dependent kinases. The three CDC25 isoforms encoding genes are submitted to alternative splicing events which generate at least two variants for CDC25A and five for both CDC25B and CDC25C. An over-expression of CDC25 was reported in several types of cancer, including breast cancer, and is often associated with a poor prognosis. Nevertheless, most of the previous studies did not address the expression of CDC25 splice variants. Here, we evaluated CDC25 spliced transcripts expression in anti-cancerous drug-sensitive and resistant breast cancer cell lines in order to identify potential breast cancer biomarkers. Methods: CDC25 splice variants mRNA levels were evaluated by semi-quantitative RT-PCR and by an original real-time RT-PCR assay. Results: CDC25 spliced transcripts are differentially expres-sed in the breast cancer cell lines studied. An up-regulation of CDC25A2 variant and an increase of the CDC25C5 / C1 ratio are associated to the multidrug-resistance in VCREMS and DOXOR breast cancer cells, compared to their sensitive counterpart cell line MCF-7. Additionally, CDC25B2 tran-script is exclusively over-expressed in VCREMS resistant cells and could therefore be involved in the development of certain type of drug resistance. Conclusions: CDC25 splice variants could represent interesting potential breast cancer prognostic biomarkers.

Background: A proliferation-inducing ligand (APRIL) is a newly-found member in the tumor necrosis factor (TNF) superfamily. Our previous studies have already confirmed that APRIL is overexpressed in pancreatic cancer tumors, however, it is not expressed or has a weak expression in normal pancreatic gland tissues. Furthermore, there is no report on serum APRIL in patients with pancreatic diseases. Herein, in order to explore the clinical implication of serum APRIL in patients with pancreatic cancer, serum APRIL, together with carcinoembryonic antigen (CEA) and carbohydrate antigen (CA)19-9, was examined. Methods: Serum APRIL was tested by ELISA in patients with pancreatic cancer. Meanwhile, two other conventional serum tumor markers, CEA and CA19-9, were measured by Elecsys 2010 Chemistry Analyzer. Results: Serum APRIL increased in patients with pancreatic cancer, which proved a positive correlation with CEA and CA19-9. When the diagnosis of benign or malignant condition was examined by one tumor marker, the sensitivity of APRIL alone (70.1%) was greater than that of CEA alone (56.7%), and the specificity of APRIL alone (85.5%) was higher than that of CA19-9 alone (83.6%). When examined by a combination of two markers, the sensitivity of the combination of APRIL and CA19-9 was the highest (88.1%), as it was compared with that of APRIL alone, CEA alone and APRIL+CEA, p<0.05. In addition, serum APRIL also correlated with the tumor stage and postoperative survival in patients with pancreatic cancer. Conclusions: Our results indicate that serum APRIL, as a potential biomarker, has a positive diagnosis and prognosis value for pancreatic cancer. Moreover, the combination assay of APRIL and CA19-9 is highly sensitive to pancreatic cancer.

Background: Malignant pleural mesothelioma (MPM) is an aggressive malignant tumor of mesothelial origin associated with asbestos exposure. MPM has a limited response to conventional chemotherapy and radiotherapy, so early diagnosis of MPM is very important. This study investigated the pleural effusion mesothelin levels in patients with MPM and compared them to those of a population with a non-malignant pleuritis or lung cancer involving malignant pleural effusion. Methods: The pleural effusion mesothelin concentrations were measured in 45 MPM patients and 53 non-MPM individuals (24 individuals with non-malignant pleural effusions and 29 individuals with lung cancer involving malignant pleural effusion). Results: This study demonstrated that patients with MPM had significantly higher pleural effusion mesothelin levels than a population with non-malignant pleuritis or lung cancer involving malignant pleural effusion. The difference in overall survival between the groups with pleural effusion mesothelin levels lower and higher than the assumed cut-off of 10 nM was significant. Conclusions: The data suggest that the pleural effusion mesothelin concentration could be useful as an aid for the diagnosis of MPM.

Background: New technologies for the early detection of tuberculosis (TB) are urgently needed. Pathological changes within an organ might be reflected in proteomic patterns in serum. The aim of the present study was to screen for the potential protein biomarkers in serum for the diagnosis of TB using proteomic fingerprint technology. Methods: Proteomic fingerprint technology combining protein chips with surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) was used to profile the serum proteins from 50 patients with TB, 25 patients with lung disease other than TB, and 25 healthy volunteers. The protein fingerprint expression of all the serum samples and the resulting profiles between TB and control groups were analyzed with the Biomarker Wizard system. Results: A total of 30 discriminating m/z peaks were detected that were related to TB (p<0.01). The model of biomarkers constructed by the Biomarker Patterns Software based on the three biomarkers (2024, 8007, and 8598 Da) generated excellent separation between the TB and control groups. The sensitivity was 84.0% and the specificity was 86.0%. Blind test data indicated a sensitivity of 80.0% and a specificity of 84.2%. Conclusions: The data suggested a potential application of SELDI-TOF MS as an effective technology to profile serum proteome, and with pattern analysis, a diagnostic model comprising three potential biomarkers was indicated to differentiate people with TB and healthy controls rapidly and precisely.

Background: Matrix metalloproteinase-7 (MMP-7) may play an important role in the development of vulnerable carotid plaque. An A-to-G transition (–181A/G) in the promoter region of MMP7 is functional in vitro by altering the transcriptional activity of the gene. The aim of this study was to investigate the association between the MMP7 –181A/G polymorphism and vulnerable carotid plaque formation. Methods: The authors enrolled 641 patients and divided them into three groups according to the carotid ultrasound examination: vulnerable plaque group (n=118), stable plaque group (n=385) and no plaque group (n=138). Traditional atherosclerosis risk factors were recorded and the MMP7 –181A/G polymorphism was genotyped by polymerase chain reaction-restriction fragment length polymorphism. Results: In the multinomial logistic regression analysis, compared to the no plaque group, no relationship between MMP7 –181AG+GG genotypes and stable carotid plaque was observed [odds ratio (OR) 1.50; p=0.239]. However, the frequency of AG+GG genotypes was significantly higher in the vulnerable plaque group (OR 2.74; p=0.008). Age was a risk factor for plaque formation, while statin treatment can reduce the prevalence of atherosclerotic plaque. Additionally, using binary logistic regression analysis between the stable and vulnerable plaque groups, this MMP7 polymor phism was associated with vulnerable plaque independently of other factors [OR 1.83; 95% confidence interval 1.08– 3.11; p=0.026]. Conclusions: The MMP7 – 181A/G polymorphism is associated with the development of vulnerable carotid plaques. Age is a risk factor for plaque formation, while statin therapy is associated with a decreased prevalence of carotid athero-matous plaques.

Background: We assessed whether glycated albumin (GA) is a useful glycemic indicator in diabetic patients with anemia who did not undergo dialysis. Methods: Hemoglobin A 1c (HbA 1c ) and GA were simultaneously measured in 370 diabetic subjects who had not undergone dialysis. The relationship between GA and HbA 1c was evaluated in patients with and without anemia. Results: GA-to-HbA 1c ratio was significantly higher (3.3±0.7 vs. 2.8±0.5, p<0.001) and the regression slope between GA and HbA 1c was steeper in diabetic patients with than in those without anemia (6.2 vs. 4.2, p<0.001). Conclusions: HbA 1c was underestimated in diabetic patients with anemia than in those without anemia, with the degree of underestimation increasing as glycemic control became poorer. GA may more accurately assess glycemic control in diabetic patients with anemia than HbA 1c .

Background: Urinary iodide concentration (UIC) is useful to evaluate nutritional iodine status. In clinical settings UIC helps to exclude blocking of the thyroid gland by excessive endogenous iodine, if diagnostic or therapeutic administration of radio-iodine is indicated. Therefore, this study established a simple test for the measurement of UIC. Methods: UIC was analyzed in urine samples of 200 patients. Samples were pre-treated at 95°C for 45 min with ammonium persulfate in a thermal cycler, followed by a photometric Sandell-Kolthoff reaction (SK) carried out in microtiter plates. For method comparison, UIC was analyzed in 30 samples by inductivity coupled plasma mass spectro-metry (ICP-MS) as a reference method. Results: Incubation conditions were optimized concerning recovery. The photometric test correlated well to the reference method (SK=0.91*ICP-MS+1, r=0.962) and presented with a functional sensitivity of 20 μg/L. UIC of patient samples ranged from <20 to 750 μg/L (median 110 μg/L); 90% of the urine samples had iodide concentrations below 210 μg/L. Conclusion: The modified SK-test takes approximately 90 min for analyses of 20 urine samples compared with 27 h for ICP-MS. The photometric test provides satisfactory results and can be performed with the basic equipment of a clinical laboratory.