bluuebunny/crossref_metadata_2025 huggingface dataset:
Glycosylation Changes on Serum Glycoproteins in Ovarian Cancer May Contribute to Disease Pathogenesis
Authors: Saldova Radka , Wormald Mark R. , Dwek Raymond A. , Rudd Pauline M.
Published: June 2013

Ovarian cancer is the most lethal of all gynaecological cancers among women. Serum CA125 is the only biomarker that is used routinely and there is a need for further complementary biomarkers both in terms of sensitivity and specificity.

N‐glycosylation changes in ovarian cancer serum glycoproteins include a decrease in galactosylation of IgG and an increase in sialyl Lewis X (SLex) on haptoglobin β‐chain, α1‐acid glycoprotein and α1‐antichymotrypsin. These changes are also present in chronic inflammation but not in malignant melanoma, where there are low levels of inflammatory processes. Acute phase proteins carrying increased amounts of SLex have an increased half‐life. Sialylation of acute phase proteins also decreases apoptosis favouring survival of cancer cells. Cancer cells produce inflammatory cytokines which influence glycosylation processing in liver parenchymal cells. Altered glycosylation of the acute phase protein transferrin plays an important role in iron homeostasis. Glycosylated transferrin and its glycans have anti‐apoptotic properties and many transferrin receptors in carcinoma could play a role in development of anaemia. Decreased galactosylation and sialylation of IgG increases the cytotoxicity of natural killer cells and complement activation via mannose‐binding lectin (MBL). Altered glycosylation of acute phase proteins and IgG suggests that cancer regulates certain pathways favouring cancer cells survival.
Association and Linkage Disequilibrium Analyses of APOE Polymorphisms in Atherosclerosis
Authors: Artieda Marta , Gañán Alberto , Cenarro Ana , García-Otín Ángel Luis , Jericó Ivonne , Civeira Fernando , Pocoví Miguel
Published: June 2013

Background: Apolipoprotein E (apo E) plays a major role in lipid metabolism, and its genetic variations have been associated with cardiovascular risk. The objective of this study was to investigate the influence of the APOE promoter (−491 A/T, −427 T/C and −219 G/T) and coding region (APOE ɛ2/ɛ3/ɛ4) polymorphisms in atherosclerosis disease by association and linkage disequilibrium analyses. Materials and methods: We analyzed these polymorphisms in a sample of 286 subjects with atherosclerosis disease: 153 subjects with atherothrombotic stroke (ATS) and 133 subjects with ischemic heart disease (IHD); and in two control groups, 103 newborns and 114 elderly subjects. Results: The ɛ4 allele was associated with more severe carotid stenosis in the ATS group, being the percentages of ɛ4 carriers 26.7% and 11.4% for the higher and lower carotid stenosis groups, respectively (p=0,066). The −491 T/T IHD subjects presented higher vessel scores than subjects A/A and A/T genotypes at that position (p=0,041), and the frequencies of −2 (5.1% versus 14.1%, p=0,060) and −427C (10.3% versus 24.4%, p=0,019) alleles were lower in IHD subjects with higher extent score versus lower extent score. The ɛ2 allele was in linkage disequilibrium with the −427C allele in all studied groups, and the −219T allele was associated with the ɛ4 allele in the IHD group. Conclusion: In summary, the ɛ2 allele was in linkage disequilibrium with the −427C allele in all studied groups, and only slight associations between the analyzed APOE polymorphisms in the promoter and in the coding region and carotid and coronary vascular disease have been observed.
Expression of EpCam and Villin in Barrett’s Esophagus and in Gastric Cardia
Authors: Anders Mario , Sarbia Mario , Grotzinger Carsten , Meining Alexander , Hofler Heinz , Wiedenmann Bertram , Rosch Thomas
Published: June 2013

In the current study we aimed to clarify the potential of EpCAM and villin as in vivo biomarkers for both Barrett esophagus (BE)‐associated neoplasia and BE versus cardiac mucosa. Immunohistochemical staining in BE with various degrees of intraepithelial neoplasia (IN), Barrett carcinoma (BC) and in normal cardiac mucosa (CM) revealed a lack of EpCam and villin in squamous esophageal epithelium. All specimens of IN and BC showed EpCam with varying staining intensities. In 57% of CM samples a weak signal was detected; the remainder displayed strong EpCam expression. Villin was found in 97% of BE specimens and in all those with IN; 37% of BC and 75% of CM specimens were also positive. We conclude that expression of EpCam and villin differs only between squamous epithelium and BE. Determination of these proteins does not allow discrimination between different degrees of neoplasia or between esophageal intestinal metaplasia and cardiac mucosa.
Human Neutrophil Peptides 1‐3 – Early Markers in Development of Colorectal Adenomas and Carcinomas
Authors: Mothes Henning , Melle Christian , Ernst Günther , Kaufmann Roland , von Eggeling Ferdinand , Settmacher Utz
Published: June 2013

Expression of Human Neutrophil Peptides (HNP) 1–3 was recently found to be associated with development of colorectal cancer. Raised defensin‐expression in tumours is believed to stem from increased infiltration of neutrophils into tumour environment.

To further specify the role of α‐defensins in tumourigenesis and progression, HNP1–3 were analyzed in colorectal adenomas and carcinomas of 87 patients and quantified in relation to cancer stage and grading. Using the ProteinChip arrays, HNP1–3 were found upregulated in both colorectal adenomas and carcinomas. By combining the array with Laser capture microscopy we were able to confirm that HNP1–3 are expressed by tumour cells but not by neutrophils or other tumour invading cells. These findings suggest that α‐defensins are more likely to contribute to tumour growth than they are to mount an effective host anti‐tumour response. However, the amount of HNP‐expression was not found to be related to tumour stage, grading, and serological tumour markers.
Circulating VEGF As a Biological Marker in Patients with Rheumatoid Arthritis? Preanalytical and Biological Variability in Healthy Persons and in Patients
Authors: Hetland Merete Lund , Christensen Ib Jarle , Lottenburger Tine , Johansen Julia Sidenius , Svendsen Mads Nordahl , Hørslev-Petersen Kim , Nielsen Lone , Jørgen Nielsen Hans
Published: June 2013

Background: Soluble vascular endothelial growth factor (VEGF) is a promising biomarker in monitoring rheumatoid arthritis (RA), but studies of pre‐analytical and biologic variability are few.

Methods: VEGF was measured by ELISA methods in serum and plasma from healthy persons and RA patients. Pre‐analytical factors were investigated. A reference interval for VEGF was established in serum and plasma from 306 healthy persons. Diurnal, day‐to‐day, week‐to‐week, long‐term variability, and impact of exercise were evaluated.

Results: Delayed processing time, room temperature, low centrifugal force and contamination of plasma with cellular elements lead to significant increases in VEGF levels, whereas storage for up to 2 years at −80°C or up to 10 freeze/thaw cycles did not affect VEGF levels. Serum VEGF levels were 7–10 fold higher than plasma VEGF levels. Reference intervals for VEGF (plasma: 45 pg/ml (range: non‐detectable to 352); serum: 328 pg/ml (53–1791)) were independent of gender and age. Short‐ and long‐term biologic variability included diurnal variation (sampling should take place after 7 AM) and impact of exercise (increased VEGF immediately after bicycling normalised within 1 hour).

Conclusions: Pre‐analytical factors and biologic variability including diurnal variation and impact of exercise should be accounted for in future studies that include circulating VEGF as a biological marker.
Protective Effect of R Allele of PON1 Gene on the Coronary Artery Disease in the Presence of Specific Genetic Background
Authors: Balcerzyk Anna , Zak Iwona , Krauze Jolanta
Published: June 2013

Background: Genetic susceptibility to CAD may be determined by polymorphic variants of genes encoding isoforms involved in the processes important in the pathogenesis of atherosclerosis, including lipids disorders. Participation of single polymorphic variants is relatively small, however its significance may increase in the presence of specific genetic or environmental background.

Aim: The aim of the study was an evaluation a possible association between single polymorphic variants of PON1, APOE, ABCA1 and PPARA genes and CAD and looking for specific multigene genotype patterns which differentiate study groups.

Materials and methods: We studied 358 subjects:178 patients with angiographically confirmed CAD and 180 blood donors without history of CAD. Polymorphisms were genotyped using PCR‐RFLP method.

Results: We observed statistically significant differences in the frequencies of R allele and R allele carriers of PON1 gene between CAD and controls. The distribution of genotypes and alleles of other analyzed genes did not differentiate the study groups, however the presence of specific genotypes (APOE– ɛ3ɛ3, ɛ3ɛ2, ABCA1 – AG, PPARA – GG) increased the protective effect of R allele.

Conclusion: The present study revealed an independent protective association between carrier‐state of PON1 R allele and CAD. This protective effect was especially strong in the presence of specific genotype arrangements of other analyzed genes.
Glyco‐biomarkers: Potential Determinants of Cellular Physiology and Pathology
Authors: Alavi Azita , Axford John S.
Published: June 2013

Once dismissed as just the icing on the cake, sugar molecules are emerging as vital components in life’s intricate machinery. Our understanding of their function within the context of the proteins and lipids to which they are attached has matured rapidly, and with it the far reaching clinical implications are becoming understood.

Recent advances in high‐throughput glycomic techniques, glyco biomarker profiling, glyco‐bioinformatics and development of increasingly sophisticated glyco‐arrays, combined with our increased understanding of the molecular details of glycosylation have facilitated the linkage between aberrant glycosylation and human diseases, and highlighted the possibility of using glyco‐biomarkers as potential determinants of disease and its progression.

The focus of this review is to give an insight into the biological significance of these glycomodifications, highlight some specific examples of glyco‐biomarkers in relation to autoimmunity and in particular rheumatoid arthritis, and to explore the exciting possibility of exploiting these for diagnostic and prognostic strategies.
Lipid Measures and Cardiovascular Disease Prediction
Authors: van Wijk Diederik F. , Stroes Erik S. G. , Kastelein John J. P.
Published: June 2013

Traditional lipid measures are the cornerstone of risk assessment and treatment goals in cardiovascular prevention. Whereas the association between total, LDL-, HDL-cholesterol and cardiovascular disease risk has been generally acknowledged, the rather poor capacity to distinguish between patients who will and those who will not develop cardiovascular disease has prompted the search for further refinement of these traditional measures. A thorough understanding of lipid metabolism is mandatory to understand recent developments in this area. After a brief overview of lipid metabolism we will discuss the epidemiological data of total, LDL- and HDL-cholesterol and focus on recent advances in measurements of these lipoproteins. In addition we will discuss the role of triglycerides and the apolipoprotein B–A-I ratio on the incidence of cardiovascular disease.
Blau Syndrome-Related CARD15/NOD2 Mutations Are Not Linked to Idiopathic Uveitis in Spanish Patients
Authors: Rodríguez-Pérez Noelia , Aguinaga-Barrilero Ana , Gorroño-Echebarría Marina B. , Pérez-Blas Mercedes , Martín-Villa José M.
Published: June 2013

Uveitis is a clinical feature of the Blau syndrome, a disease linked to CARD15 (also referred to as NOD2) mutations. Three main mutations in this gene (R334W, R334Q and L469F) have been reported as Blau syndrome risk factors, a disease that manifests uveitis as one of its clinical features. However, little is known on the involvement of this gene in idiopathic uveitis. We thus sought to determine the frequency of these Blau-related CARD15 mutations in a cohort of Spanish patients with idiopathic uveitis. To this aim, 110 patients with idiopathic uveitis, followed at the Department of Ophtalmology of a tertiary hospital (Hospital Universitario Alcalá de Henares, Madrid. Spain) were enrolled. As a control population, 104 healthy subjects were used. DNA was extracted from blood samples and the Blau-related CARD15 mutations were analysed either by PCR-RFLP or direct DNA sequencing. None of the mutations studied was found in any of the individuals tested, whether diseased or healthy. It seems thus that Blau syndrome-related CARD15 mutations are not involved in idiopathic uveitis, a finding which allows us to suggest that the genetic aetiology of the idiopathic uveitis or the Blau-associated uveitis is different.
Association of Single Nucleotide Polymorphisms in CYP1B1 and COMT Genes with Breast Cancer Susceptibility in Indian Women
Authors: Yadav Sharawan , Singhal Naveen Kumar , Singh Virendra , Rastogi Neeraj , Srivastava Pramod Kumar , Singh Mahendra Pratap
Published: June 2013

Cytochrome P450 1B1 (CYP1B1) and catechol-$O$-methyltransferase (COMT) enzymes play critical roles in estrogen metabolism. Alterations in the catalytic activity of CYP1B1 and COMT enzymes have been found associated with altered breast cancer risk in postmenopausal women in many populations. The substitution of leucine (Leu) to valine (Val) at codon 432 increases the catalytic activity of CYP1B1, however, substitution of Val to methionine (Met) at codon 158 decreases the catalytic activity of COMT. The present study was performed to evaluate the associations of CYP1B1 Leu432Val and/or COMT Val158Met polymorphisms with total, premenopausal and postmenopausal breast cancer risks in Indian women. COMT and CYP1B1 polymorphisms in controls and breast cancer patients were analyzed employing polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) followed by gel electrophoresis. Although CYP1B1 and COMT genotypes did not exhibit statistically significant association with breast cancer risks when analyzed individually, COMT wild type (Val158Val) in combination with CYP1B1 heterozygous variant (Leu432Val) [OR: 0.21; 95% CI (0.05–0.82), p value; 0.021] and COMT heterozygous variant (Val158Met) in combination with CYP1B1 wild type (Leu432Leu) [OR: 0.29; 95% CI (0.08–0.96), p value; 0.042] showed significant protective association with premenopausal breast cancer risk. The results demonstrate that CYP1B1 wild type in combination with COMT heterozygous or their inverse combination offer protection against breast cancer in premenopausal Indian women.
Virological, Immune and Host genetics Markers in the Control of HIV Infection
Authors: Mothe Beatriz , Ibarrondo Javier , Llano Anuska , Brander Christian
Published: June 2013

HIV infection, if left untreated, leads in most cases to the development of wide immune deterioration, opportunistic infections and eventually AIDS and death. The identification of individuals who despite persisting infection show no or few signs of HIV disease progression has spurred hopes that an effective HIV vaccine could be attainable. The design of such a vaccine will greatly depend on the precise definition of disease markers, host genetic and immune characteristics that mediate relativein vivocontrol of this virus. Accordingly, a number of viral factors and host genetic characteristics have been shown to play a crucial role in the control of HIV disease by delaying progression to AIDS or even preventing infection. There is also an improved understanding of humoral and cellular immune responses in terms of specificity, functional repertoire, longevity and tissue distribution and their ability to contain HIV replication. However, the definition of good immune correlates unequivocally and causally associated with protection or disease progression remains elusive. Here we review work on viral factors, host genetic markers and immunological determinants that have been identified in individuals with superior control of HIV infection or in subjects who remain uninfected despite frequent exposure to the viral pathogen.
Evaluating the Prognostic Value of New Cardiovascular Biomarkers
Authors: Wood Angela M. , Greenland Philip
Published: June 2013

New predictors of cardiovascular outcomes are widely sought in research settings, and predictive tests are commonly recommended for routine use in cardiovascular clinical care. A number of multivariable scoring systems are in use around the world for assessment of a patient’s risk. While such scoring systems are often recommended for clinical use in medical practice guidelines, their actual use in medical care falls short of recommendations. Limitations in the predictive capacity of existing predictive models are recognized, including lack of predictive accuracy, lack of ability to separate those who develop events from those who do not, and risks and costs of the testing modalities. Biomarker research is actively developing new testing strategies trying to improve upon current approaches, but it is often unclear how to assess the incremental prognostic information that a new test provides. In this report, we discuss the statistical approaches that can be used to evaluate additive predictive value of new tests. We also consider clinical research examples to put this information into a practical context.
Expression of ICAM1 and VCAM1 Serum Levels in Rheumatoid Arthritis Clinical Activity. Association with Genetic Polymorphisms
Authors: Navarro-Hernández Rosa Elena , Oregon-Romero Edith , Vázquez-Del Mercado Mónica , Rangel-Villalobos Héctor , Palafox-Sánchez Claudia Azucena , Muñoz-Valle José Francisco
Published: June 2013

To investigate the association of sICAM-1 and sVCAM-1 with ICAM1 721G>A and VCAM1 1238G>C polymorphisms and rheumatoid arthritis (RA) clinical activity, sixty RA patients and 60 healthy non-related subjects (HS) matched for age and sex were recruited. Soluble adhesion molecules were determined by ELISA technique. Rheumatoid factor (RF), C reactive protein (CRP) and the erythrocyte sedimentation rate (ESR) were measured by routine methods. Disability and clinical activity was measured with Spanish-HAQ-DI and DAS28 scores, respectively. The ICAM1 and VCAM1 polymorphism were identified using the PCR-RFLP procedure. Inter-group comparison showed increased levels of sICAM-1 and sVCAM-1 in RA patients (284 and 481 ng/mL) versus HS (132 and 280 ng/mL); in the RA group, significant correlations between sVCAM-1 and RF (r= 0.402), ESR (r= 0.426), Spanish-HAQ-DI (r= 0.276), and DAS28 (r= 0.342) were found, whereas sICAM-1 only correlated with RF (r= 0.445). In RA patients, a significant association with the 721A allele of ICAM1 polymorphism (p= 0.04), was found. In addition, the allele impact (G/A+A/A) of this polymorphism was confirmed, (p= 0.038, OR = 2.3, C.I. 1.1–5.0). sVCAM-1 and sICAM-1 serum levels reflected the clinical status in RA, independently of the ICAM1 and VCAM1 polymorphism. However, the ICAM1 721A allele could be a genetic marker to RA susceptibility.
Metabolic Background Determines the Importance ofNOS3Polymorphisms in Restenosis after Percutaneous Coronary Intervention: A Study in Patients with and without the Metabolic Syndrome
Authors: Pons Douwe , Monraats Pascalle S. , Zwinderman Aeilko H. , de Maat Moniek P. M. , Doevendans Pieter A. F. M. , de Winter Robbert J. , Tio René A. , Waltenberger Johannes , Jukema J. Wouter
Published: June 2013

Variation in theNOS3gene has been related to the development of restenosis. The Glu298Asp polymorphism has previously been investigated for its effect on NO levels and the development of restenosis. However, the variability of findings gave rise to the hypothesis that the functional significance of this polymorphism may only become manifest under conditions of endothelial dysfunction. Since patients with the metabolic syndrome are known to have endothelial dysfunction, we aimed to investigate if the significance ofNOS3polymorphisms may depend on the presence of the metabolic syndrome.

We examined the impact of the -949 A/G, the -716 C/T and the Glu298Asp polymorphisms in theNOS3gene on the risk of clinical restenosis in a previously described subpopulation of the GENDER-study, a multicenter prospective study design that enrolled consecutive patients after successful PCI. This subpopulation consisted of 901 patients of whom sufficient data were available to establish absence or presence of the metabolic syndrome. Of these patients, 448 had the metabolic syndrome. Clinical restenosis, defined as target vessel revascularization (TVR), was the primary endpoint.

We demonstrated that the minor -949G, -716T and 298Asp alleles were associated with a significantly increased risk of TVR in patients with the metabolic syndrome (HR: 1.58 [95%CI: 1.04–2.40], HR: 1.95 [95%CI: 1.02–3.70] and HR: 1.67 [95%CI: 1.09–2.54], respectively). In the group without the metabolic syndrome we observed no association between the three polymorphisms and TVR.
Serum Surfactant Protein D during Acute Exacerbations of Chronic Obstructive Pulmonary Disease
Authors: Shakoori Tania Ahmed , Sin Don D. , Ghafoor Farkhanda , Bashir Saira , Bokhari S. Nazim Hussain
Published: June 2013

Background: There is a paucity of lung specific biomarkers to diagnose exacerbations of chronic obstructive pulmonary disease (COPD) and to track their progression. Surfactant protein D (SP-D) is a pulmonary collectin regulating the innate immunity of the lung and its serum expression is perturbed in COPD. However, it is not known whether serum levels change during exacerbations. We sought to determine whether serum SP-D levels are raised in COPD exacerbations.

Objectives: To determine whether or not patients with exacerbations have elevated serum SP-D levels compared with asymptomatic controls, stable disease.

Study design: case control study.

Methods: We measured serum SP-D levels from patients with stable COPD (n= 14), patients experiencing acute exacerbations (n= 13) and in control subjects (n= 54) using a specific immunoassay and compared the levels using analysis of variance.

Results: Serum SP-D levels were significantly increased in patients who experienced an acute exacerbation (227 ± 120 ng/mL) compared to patients with stable disease (151 ± 83 ng/mL) or control subjects (128 ± 65 ng/mL;p= 0.003). Serum SP-D levels were also found to be inversely related to various lung function parameters including FEV1/FVC% predicted.

Conclusions: Our study suggests that serum SP-D levels are increased in patients during exacerbations and may be a potential diagnostic biomarker for COPD exacerbations.
Natriuretic Peptides~: Prediction of Cardiovascular Disease in the General Population and High Risk Populations
Authors: Hildebrandt Per
Published: June 2013

The natriuretic peptides, especially the B-type peptide (BNP) and its inactive split-product N-terminal proBNP (Nt-proBNP) are increasingly used in screening for heart failure, primarily with reduced systolic function, in patients with symptoms suggestive of heart failure, as well in the stable (General Practitioner) setting as in the acute setting.

Supporting this use is a very strong prognostic value of the natriuretic peptides. This has been shown in as well heart failure as acute coronary syndromes, but also in the general population and in high-risk groups as patients with diabetes, hypertension and coronary artery disease.

This has of course raised interest for the use of the natriuretic peptides as a risk marker and for screening for heart failure with reduced systolic function in these populations. In symptomatic persons and in high risk populations, the natriuretic peptides have demonstrated a high sensitivity for ruling out the disease, if the right decision limits are choosen. Thus the number of normal echocardiographies can be reduced.

More recently, the use in screening asymptomatic persons for left ventricular systolic dysfunction has gained more interest. In the unselected population, screening would probably not be cost-effective. In populations with a higher pre-test probability for heart failure, as patients with diabetes, hypertension and stable coronary artery disease, screening would presumably be more cost-effective, but evidence for the use in this setting is still lacking.
Prognostic Biomarkers in Individuals with Prevalent Coronary Heart Disease
Authors: Halim Sharif A. , Newby L. Kristin
Published: June 2013

Coronary disease is the leading killer of individuals worldwide and a leading cause of healthcare expenditure. On a global scale, ischemic heart disease kills over 6 million individuals each year and is projected by the World Health Organization to be the greatest single-disease cause of death worldwide by an increasing margin into 2030. Nearly 17 million Americans (7.6% of the population) have prevalent coronary heart disease, 8 million of whom have had a prior myocardial infarction. It is estimated that in 2009, 550,000 will die from coronary heart disease in the United States and that the direct and indirect costs from treating coronary heart disease will exceed $165 billion. Although patients with known coronary artery disease are among the highest risk patients for future cardiac events, not all patients with coronary disease will have an ischemic event (first or recurrent). Determining which of these patients will have an ischemic event is critical to the concept of personalized cardiovascular care. Increasingly, biomarkers that can be readily assayed from blood or other body fluids will be critical to risk stratification and effective application of secondary prevention strategies, just as they have played an increasingly prominent role in risk stratification of acute coronary syndrome patients.
Rapid and Accurate Approach for Screening of Microsatellite Unstable Tumours Using Quasimonomorphic Mononucleotide Repeats and Denaturating High Performance Liquid Chromatography (DHPLC)
Authors: Berginc Gašper , Glavač Damjan
Published: June 2013

MSI analysis is becoming increasingly important for the detection of both hereditary non-polyposis colorectal cancer and sporadic primary colorectal tumours with MSI high phenotype. The Bethesda panel of five microsatellite markers has been proposed to provide uniform criteria for MSI analysis. Here we report on an MSI analysis approach using quasimonomorphic mononucleotide repeats and denaturating high performance liquid chromatography (DHPLC). We analysed 595 newly diagnosed colorectal tumours and 145 normal samples. Microsatellite markers BAT-25, BAT-26, NR-21, NR-22, and NR-27 were amplified in multiplex reaction and analysed using DHPLC and capillary electrophoresis (CE). DHPLC conditions for analysis of MSI multiplex assay were evaluated and tested. Analysis and cross-examination of the results obtained from 96 samples using DHPLC and capillary electrophoresis showed the same sensitivity and specificity of the two approaches for detecting MSI-H tumours. Using our new approach we showed that the tested markers are quasimonomorphic in a Slovenian population, with frequencies of polymorphisms 0.07%, 1.4%, 2.1%, 1.4%, and 1.4% for BAT-25, BAT-26, NR-21, NR-22, and NR-27, respectively. Forty-three (7.2%) new MSI-H tumours were identified, of which 84% showed instability in all 5 tested markers. Overall, we developed a high-throughput, robust, accurate and cost-effective approach for the detection of MSI-H tumours.
HLA Haplotype A33-B58-Cw10 May Modulate Radiographic Development of Bamboo Spine in Taiwanese Patients with Primary Ankylosing Spondylitis
Authors: Lu Ming-Chi , Yang Kuo-Liang , Huang Kuang-Yung , Tung Chien-Hsueh , Liu Su-Qin , Lai Ning-Sheng
Published: June 2013

Objective: To evaluate the possible relationship between HLA alleles and bony ankylosis of the spine (bamboo spine) in Taiwanese patients with ankylosing spondylitis (AS).

Methods: A small cohort of HLA-B27 positive AS patients was conducted to analyze the effects of alleles and haplotypes on the development of bamboo spine. DNA typing of HLA class I and class II genes were performed by SSP method on primary ankylosing spondylitis patients with bamboo spine (n= 84) and spinal enthesopathy controls (n= 228). Odds ratios with 95% confidence intervals andPvalue were estimated. Determination of the most probable HLA haplotypes on all patients were constructed by comparison of the alleles carried by each patient with the HLA haplotype database established in Taiwanese population studies using homozygosity approach [1] and by expectation-maximum algorithm [2].

Results: Allele frequencies of HLA A33, B58, Cw10, DR4, DR17 and DQ2 were significantly lower in bamboo patients as compared to non-bamboo controls. In contrast, allele frequencies of A24, B54, Cw15, DR11 and DR14 were significantly higher in bamboo patients. Less remarkably, high frequencies of B39, B51, Cw1 and Cw2 alleles were also noted in bamboo patients. Considering linkage disequilibria of alleles in haplotypes, HLA-A11-B27-Cw12 was the most common haplotype in both bamboo and non-bamboo groups (95.23% vs. 91.22%, respectively,P= 0.238). Haplotypes A33-B58-Cw10, A33-B58-Cw10-DR13 and A33-B58-Cw10-DR17 were significantly lower in bamboo patients as compared to those in controls (P< 0.001,P= 0.001,P= 0.002, respectively).

Conclusion: Haplotypes A33-B58-Cw10, A33-B58-Cw10-DR13 and A33-B58-Cw10-DR17 showed a strong association with bamboo spine in Taiwanese AS patients. Detection of such haplotypes might be a great aid in the management of patients with the disease.
Frequency ofP16INK4aandP14ARFGenes Methylation and Its Impact on Bladder Cancer Cases in North Indian Population
Authors: Ali Hosseini Seyed , Sobti Ranbir Chander , Malekzadeh Kianoosh , Singh Shrawan Kumar , Joshi Kusum
Published: June 2013

Introduction:Amongst the genitourinary cancers, carcinoma of the urinary bladder is one of the leading causes of death in India. Hypermethylation of the CpG islands of gene promoter is one of the earliest and most frequent epigenetic alterations leading to cancer as well as in its development. Several studies have suggested that tumour suppressor genes play a key role in the development of cancer. Methylation in theCDKN2Ahas been associated with various malignant diseases, but information with respect to urinary bladder cancer is lacking in north Indian population.

Materials and methods:We analyzed the methylation ofP16INK4aandP14ARFin 80 tissues and matched blood samples of patients suffering from bladder cancer and 80 blood samples of cancer-free individuals by MS-PCR.

Results:In tissue and matched blood samples of bladder cancer patients, the incidence ofP14ARFhypermethylation significantly increased (OR = 0.31, 95%CI = 0.12–0.8,P= 0.01) and (OR = 0.0, 95%CI=0.0–0.62,P= 0.006) respectively with an increase in age. Clinicopathological analysis revealed that P14ARF hypermethylation in tissue and blood samples was significantly associated with invasive stage (≥ T2) (OR = 0.21, 95%CI = 0.08–0.51,P= 0.0002) and (OR = 0.09, 95%CI = 0.03–0.37,P= 0.00001) respectively. Muscle invasive tumour stage (≥T2) showed significant association with increased risk of P16INK4α promoter hypermethylation in tissue and blood samples of patients (OR = 0.38, 95%CI = 0.17–0.82,P= 0.01) and (OR = 0.13, 95%CI = 0.05–0.36,P= 0.00005) respectively.

Conclusion:These results suggest that the CpG island hypermethylation status of the defined panel of genes may be a useful biomarker in patients suffering from bladder cancer.
Monitoring Proteins and Protein Networks Using Reverse Phase Protein Arrays
Authors: Wilson Bridget , Liotta Lance A. , Petricoin III Emanuel
Published: June 2013

Recent advances in high throughput, high content “omic” technologies coupled with clinical information has lead to the expectation that the complexity of the molecular information generated will lead to more robust scientific research as well as the expectation that overarching therapeutic approaches will be patient-tailored to the underlying specific molecular defects of the disease. As disease understanding progresses and more therapeutics, which predominately target proteins, are developed there is a need to more confidently determine the protein signaling events that can be correlated with drug response since the deranged protein signaling networks are often the drug target itself. In this environment, the Reverse Phase Protein Microarray (RPMA) can be utilized to address the needs of both clinical screening and disease understanding through its ability to provide an unmatched functional and highly multiplexed signaling network level mapping of ongoing signaling activation, coupled with the ability of the platform to provide this information reproducibly from a tiny needle biopsy specimen or fine needle aspirate. This platform has now been utilized for biomarker discovery/validation and advancements in disease understanding both in the clinic and at the bench in the fields of cancer, liver disease, immunological disorders, and bacterial infection.
Association of Hypoadiponectinemia with Hypoglutathionemia in NAFLD Subjects with and without Type 2 Diabetes
Authors: Sandhya Narasimhan , Gokulakrishnan Kuppan , Ravikumar Radhakrishnan , Mohan Viswanathan , Balasubramanyam Muthuswamy
Published: June 2013

Objective: The aim of this study is to measure the extent of oxidative stress and to see whether it has any correlation to changes in adiponectin levels in NAFLD subjects with and without Type 2 diabetes.

Methods: Subjects recruited from the Chennai Urban Rural Epidemiology Study comprise of 1: Normal Glucose Tolerance (NGT) subjects without NAFLD; 2: NGT with NAFLD; 3: Type 2 Diabetic patients [T2DM] without NAFLD and 4: T2DM with NAFLD. Thiobarbituric acid reactive substances (TBARS), protein carbonyl (PCO), glutathione and adiponectin levels were measured by standard methods. Ultrasound of the liver was used to diagnose NAFLD.

Results: T2DM subjects with NAFLD had significantly (p< 0.001) higher levels of thiobarbituric acid reactive substances (TBARS) and protein carbonyls (PCO) but lower (p< 0.001) GSH/GSSG ratio and adiponectin levels compared to other three groups. The association of hypoadiponectinemia withNAFLD/Type 2 diabetes was significant even after adjusting for age, gender and BMI, but lost when adjusted for parameters of oxidative stress. While palmitate significantly reduced GSH/GSSG ratio in hepatocytes, addition of exogenous recombinant adiponectin restored the GSH/GSSG ratio comparable to those of untreated cells.

Conclusion: There exists an association of hypoglutathionemia and hypoadiponectinemia in subjects with NAFLD and/or T2DM. In addition to the known beneficial effects, out study also exposes the antioxidant nature of adiponectin.
Platelet Markers Correlate with Glycemic Indices in Diabetic, but Not Diabetic-Myelodysplastic Patients with Normal Platelet Count
Authors: Dalamaga Maria , Karmaniolas Konstantinos , Lekka Antigoni , Antonakos George , Thrasyvoulides Apollon , Papadavid Evangelia , Spanos Nikolaos , Dionyssiou-Asteriou Amalia
Published: June 2013

Background: Altered thrombocyte morphology and function have been reported in patients with diabetes mellitus (DM) type 2. The aim of the present study was to determine the associations between platelet morphology markers and hemoglobin A1C (HbA1c), fasting glucose (FG), hypertension and coronary heart disease (CHD) in patients with myelodysplastic syndromes (MDS) and DM, in patients with DM and in controls.

Methods: This cross-sectional study included 30 cases with primary MDS with normal platelet count and non-insulin dependent diabetes, 30 non-insulin dependent diabetic patients and 30 non-diabetic, non-MDS controls matched on age and gender. Results: After adjusting for body mass index, platelet number, CHD and hypertension, HbA1cand FG were significant predictors of mean platelet volume (MPV) and platelet distribution width (PDW) in diabetic patients. There was no correlation between platelet parameters and HbA1cor FG in diabetic MDS patients. In controls, FG and hypertension predicted significant differences in platelet morphology. Platelet count correlated with platelet morphology in diabetic MDS and control groups, but not in diabetics.

Conclusions: MPV and PDW are associated with glycemic indices in diabetic patients but not in diabetic MDS patients with normal platelet counts. Non-diabetic controls also exhibit FG related changes in platelet morphology. This suggests other factors inherent to bone marrow dysplasia, platelet turnover and biochemistry, or vascular environment affect platelet morphology in diabetic MDS patients even with normal platelet count. Platelet morphology in this population may be an early marker for myelodysplasia. These findings also support platelet morphology change as a marker for elevated macrovascular disease risk.
The PCA and LDA Analysis on the Differential Expression of Proteins in Breast Cancer
Authors: Liang Seng , Singh Manjit , Dharmaraj Saravanan , Gam Lay-Harn
Published: June 2013

Breast cancer is a leading cause of mortality in women. In Malaysia, it is the most common cancer to affect women. The most common form of breast cancer is infiltrating ductal carcinoma (IDC). A proteomic approach was undertaken to identify protein profile changes between cancerous and normal breast tissues from 18 patients. Two protein extracts; aqueous soluble and membrane associated protein extracts were studied. Thirty four differentially expressed proteins were identified. The intensities of the proteins were used as variables in PCA and reduced data of six principal components (PC) were subjected to LDA in order to evaluate the potential of these proteins as collective biomarkers for breast cancer. The protein intensities of SEC13-like 1 (isoform b) and calreticulin contributed the most to the first PC while the protein intensities of fibrinogen beta chain precursor and ATP synthase D chain contributed the most to the second PC. Transthyretin precursor and apolipoprotein A-1 precursor contributed the most to the third PC. The results of LDA indicated good classification of samples into normal and cancerous types when the first 6 PCs were used as the variables. The percentage of correct classification was 91.7% for the originally grouped tissue samples and 88.9% for cross-validated samples.
Immunological Markers of Lung Disease Due to Non-Tuberculous Mycobacteria
Authors: Lim Andrew , Allison Caris , Tan Dino Bee Aik , Oliver Ben , Price Patricia , Waterer Grant
Published: June 2013

Lung disease due to nontuberculous mycobacteria (NTM) is a poorly understood condition that is difficult to treat. Treatment remains problematic as few tools are available to help clinicians monitor disease progression or predict treatment outcome. In this study, plasma levels of several inflammatory molecules and the frequency of circulating T cell subsets were measured in patients with NTM lung disease and known treatment status, and compared with their adult offspring and with unrelated healthy controls. Plasma levels of the chemokine CXCL10 and IL18 were assessed for associations with treatment efficacy. CXCL10 was higher in patients than adult offspring (p < 0.001) and unrelated controls (p< 0.001). Plasma CXCL10 was also lower in patients who responded well to therapy or who controlled their infection without requiring therapy, when compared to patients who did not respond to therapy (p= 0.03). Frequencies of activated (HLADR+) CD4+T cells were higher in patients than adult offspring (p< 0.001) and unrelated controls (p< 0.05), with the highest frequencies in individuals who had completed at least 6 months of treatment. Frequencies of activated (CD38+) CD8+T cells in most treatment responders were similar to unrelated controls. Low plasma levels of CXCL10 may reflect successful control of NTM lung disease with or without therapy. Compared with responders, patients who responded poorly to treatment generally had higher plasma levels of CXCL10 and IL18, and higher frequencies of activated CD8+T cells.
Interleukin-6 Gene Promoter Polymorphisms and Cardiovascular Risk Factors. A family study
Authors: Guzmán-Guzmán Iris Paola , Muñoz-Valle José Francisco , Flores-Alfaro Eugenia , Salgado-Goytia Lorenzo , Salgado-Bernabé Aralia Berenice , Parra-Rojas Isela
Published: June 2013

Interleukin-6 (IL-6) is a cytokine involved in inflammatory process, as well as in glucose and lipid metabolism. Several studies of the biological relevance of IL-6 gene polymorphisms have indicated a relationship with cardiovascular disease. The aim of this study was to assess whether the –174 G/C and –572 G/C of IL-6 gene polymorphisms are associated with cardiovascular risk factors in Mexican families. Ninety members of 30 Mexican families, in which an index case (proband) had obesity, were included in the study. We evaluated the body composition by bioelectrical impedance. Peripheral blood samples were collected to determine biochemical and hematological parameters. High sensitivity C- reactive protein levels were measurement for nephelometric analysis. Screening for both polymorphisms studied was performed by PCR-RFLP. In the parents, both polymorphisms were in Hardy-Weinberg's equilibrium. The genotypes –174 GC/CC were associated with T2D (OR = 1.23, IC95%1.01–1.5) and highest levels of hsCRP (p= 0.02), whereas genotype –572 GG was associated with T2D (OR = 1.24, IC95%1.04–1.47) with an inflammatory state determined by the increase in the leukocyte count (OR = 1.24, IC95%1.02–1.51). The genotypes –174 GC/CC and –572 GG may confer susceptibility for the development of subclinical inflammation and type 2 diabetes in Mexican families.
High Expression of Cathepsin D in Non-Hodgkin’s Lymphomas Negatively Impacts on Clinical Outcome
Authors: Nicotra Giuseppina , Manfroi Federica , Follo Carlo , Castino Roberta , Fusco Nicola , Peracchio Claudia , Kerim Simonetta , Valente Guido , Isidoro Ciro
Published: June 2013

The lysosomal protease Cathepsin D (CD) has been implicated in the homeostasis of lymphatic tissues. We investigated whether the level of CD expression influences the progression and the clinical outcome in Non-Hodgkin’s Lymphomas (NHLs). The expression of CD was assessed by immunohistochemistry and immunofluorescence in biopsies of Diffuse Large B Cell Lymphomas (DLBCL, 35 cases), Follicular Lymphomas (FL, 9 cases of grade I-II plus 14 cases of grade IIIB), Chronic Lymphocytic Leukaemias (CLL, 17 cases) and Peripheral T-cell Lymphomas (PTCL, 5 cases). CD staining showed a cytoplasmic punctate pattern compatible with its lysosomal localization. Based on the level of CD expression and the proportion of positive cells, lymphomas were classified as ‘low expressing’ (< 20% of tumor cells) or ‘highly expressing’ (≥ 20% of tumor cells). Lymphomas highly expressing CD were associated with a worse stage (III-IV) at diagnosis (31/34 cases;p= 0.002) and with a poor clinical outcome (i.e., partial remission and death; 28/34 cases;p= 0.03). In the subgroup of aggressive/high grade of malignancy lymphomas (i.e., DLBCL, FL IIIB and PTCL), the Kaplan-Meier curve revealed a very low cumulative overall survival probability (~20% at 5 year) for patients bearing a NHL with > 40% CD-positive cells compared to that of patients bearing a NHL with < 20% CD-positive cells (~70% at 5 year). This correlation was statistically significant (log-rank test,p= 0.01). In Cox multivariate analysis CD failed to be a prognosticator independent of pathologic stage, though the hazard ratio confirmed the association of low expression with a better survival probability. These data indicate that the presence of a high percentage of CD-positive tumor cells negatively reflects on the progression of NHLs.
14-3-3 Sigma And p53 Expression in Gastric Cancer and Its Clinical Applications
Authors: Mühlmann Gilbert , Öfner Dietmar , Zitt Matthias , Müller Hannes M. , Maier Hans , Moser Patrizia , Schmid Kurt W. , Zitt Marion , Amberger Albert
Published: June 2013

14-3-3 sigma (σ) induces G2 arrest enabling the repair of damaged DNA. The function of 14-3-3 σ is frequently lost in tumor cells, indicating a potential tumor suppressor function. The purpose of this study was to evaluate the prognostic value of 14-3-3 σ expression in human gastric cancer. 14-3-3 σ expression was analyzed by immunohistochemistry in 157 tumor samples of patients, who underwent resection for gastric cancer. Since 14-3-3 σ is involved in the p53 network, p53 expression was detected in parallel and correlated with 14-3-3 σ. 14-3-3 σ was found to be overexpressed in 75 (47.8%) of 157 cases, the overexpression rate of p53 protein was 27.4%. 14-3-3 σ overexpression was statistically significantly associated with pT-stage (p=0.041) pN-stage (p=0.015) and UICC-stage (p=0.019) and showed a borderline significance with Lauren classification (p=0.057). Univariate survival calculations revealed a coexistent 14-3-3 σ and p53 overexpression as a significant predictor of disease-free survival. Multivariate analysis did not unfold 14-3-3 as an independent prognostic factor for disease-free survival and overall survival. Concomitant 14-3-3 σ and p53 overexpression in tumor cells of patients with gastric cancer identifies a population of patients with relatively unfavorable prognosis.
Serum Markers of Liver Fibrosis: Combining the BIPED Classification and the Neo-Epitope Approach in the Development of New Biomarkers
Authors: Veidal Sanne Skovgård , Bay-Jensen Anne-Christine , Tougas Gervais , Karsdal Morten Asser , Vainer Ben
Published: June 2013

Background:Fibrosis is a central histological feature of chronic liver diseases and is characterized by the accumulation and reorganization of the extracellular matrix. The gold standard for assessment of fibrosis is histological evaluation of a percutaneous liver biopsy. Albeit a considerable effort have been invested in finding alternative non-invasive approaches, these have not been sufficiently succesfull to replace biopsy assessment.

Aim:To identify the extracellular matrix proteins of interest, that as protein degradation fragments produced during extracellular matrix metabolism neo-epitopes, may be targeted for novel biochemical marker development in fibrosis. We used the recently proposed BIPED system (Burden of disease,Investigative,Prognostic,Efficacy andDiagnostic) to characterise present serological markers.

Methods:Pubmed was search for keywords; Liver fibrosis, neo-epitopes, biomarkers, clinical trail, extra cellular matrix, protease, degradation, fragment.

Results and Conclusion:Implementation of BIPED categorization in the development and validation of fibrosis biomarkers to simplify and standardize the use of existing and future biomarkers seems advantageous. In addition, a systematic use of the neo-epitope approach, i.e. the quantification of peptide epitopes generated from enzymatic cleavage of proteins during extracellular remodeling, may prove productive in the quest to find new markers of liver fibrosis.
The Differential Expression of Aqueous Soluble Proteins in Breast Normal and Cancerous Tissues in Relation to Ethnicity of the Patients; Chinese, Malay and Indian
Authors: Liang Seng , Singh Manjit , Gam Lay-Harn
Published: June 2013

Female breast cancer is one of the leading causes of female mortality worldwide. In Malaysia, breast cancer is the most commonly diagnosed cancer in women. Of the women in Malaysia, the Chinese have the highest number of breast cancer cases, followed by the Indian and the Malay. The most common type of breast cancer is infiltrating ductal carcinoma (IDC). A proteomic approach was applied in this study to identify changes in the protein profile of cancerous tissues compared with normal tissues from 18 patients; 8 Chinese, 6 Malay and 4 Indian were analysed. Twenty-four differentially expressed hydrophilic proteins were identified. We evaluated the potential of these proteins as biomarkers for infiltrating ductal carcinoma based on their ethnic-specific expressions. Three of the upregulated proteins, calreticulin, 14-3-3 protein zeta and 14-3-3 protein eta, were found to be expressed at a significantly higher level in the cancerous breast tissues when compared with the normal tissues in cases of infiltrating ductal carcinoma. The upregulation in expression was particularly dominant in the Malay cohort.