bluuebunny/crossref_metadata_2025 huggingface dataset:
Genetic Variants of Surfactant Proteins A, B, C, and D in Bronchopulmonary Dysplasia
Authors: Pavlovic J. , Papagaroufalis C. , Xanthou M. , Liu W. , Fan R. , Thomas N. J. , Apostolidou I. , Papathoma E. , Megaloyianni E. , DiAngelo S. , Floros J.
Published: June 2013

BPD_28D (O2 dependency at 28 days of life) and BPD_36W (O2 dependency at 36 wks post‐menstrual age) are diseases of prematurely born infants exposed to mechanical ventilation and/or oxygen supplementation. In order to determine whether genetic variants of surfactant proteins (SPs‐A, B, C, and D) and SP‐B‐linked microsatellite markers are risk factors in BPD, we performed a family based association study using a Greek study group of 71 neonates (<30 wks gestational age) from 60 families with, 52 BPD_28D and 19 BPD_36W, affected infants. Genotyping was performed using newly designed pyrosequencing assays and previously published methods. Associations between genetic variants of SPs and BPD subgroups were determined using Transmission Disequilibrium Test (TDT) and Family Based Association Test (FBAT). Significant associations (p ≤ 0.01) were observed for alleles of SP‐B and SP‐B‐linked microsatellite markers, and haplotypes of SP‐A, SP‐D, and SP‐B. Specifically, allele B‐18_C associated with susceptibility in BPD_36W. Microsatellite marker AAGG_6 associated with susceptibility in BPD_28D/36W group. Haplotype analysis revealed ten susceptibility and one protective haplotypes for SP‐B and SP‐B‐linked microsatellite markers and two SP‐A‐SP‐D protective haplotypes. The data indicate that SP loci are linked to BPD. Studies in different study groups and/or of larger sample size are warranted to confirm these observations and delineate genetic background of BPD subgroups.
TLR‐4 and CD14 Polymorphisms in Respiratory Syncytial Virus Associated Disease
Authors: Puthothu Beena , Forster Johannes , Heinzmann Andrea , Krueger Marcus
Published: June 2013

Respiratory syncytial virus (RSV) is the most common viral respiratory pathogen during infancy world wide. It induces innate and adaptive immune response in host cells. The toll like receptor 4 (TLR4)/CD14 complex is particularly important for the initiation of an innate immune response to RSV. Thus we were interested whether an association exists between severe RSV associated diseases and polymorphisms within TLR4 and CD14.

We genotyped the CD14 promotor polymorphism ‐C159T and the two common TLR4 amino acid variants (D259G, and T359I) in 131 infants with severe RSV associated diseases and 270 controls. Statistical analyses of single polymorphisms made use of the Armitage’s trend test, haplotypes were calculated by FAMHAP, FASTEHPLUS and Arlequin.

All polymorphisms were in Hardy Weinberg Equilibrium. We found marginal association between amino acid exchange D259G in TLR4 with RSV infection p = 0.0545). Furthermore, haplotypes analysis of the two TLR4 polymorphisms by three independent programs revealed association of haplotypes with severe RSV infection (p ≤ 0.0010). In contrast, the promotor polymorphism within CD14 was not associated with susceptibility to RSV disease. We conclude from our study, that TLR4 polymorphisms, and particularly the haplotypes, may influence the genetic predisposition to severe RSV infection.
Remarkable Variation in the Informativeness of RFLP Markers Linked to Hemophilia B Locus in Indian Population Groups: Implication in the Strategy for Carrier Detection
Authors: Mukherjee S. , Saha A. , Kumar P. Senthil , Chandak G. R. , Majumder P. P. , Ray K.
Published: June 2013

Hemophilia B, an X‐linked recessive bleeding disorder, is caused by heterogeneous mutations in the factor IX (F9) gene. Hence, carriers of the disease are usually detected by F9 gene linked RFLP analysis. We aimed to test a set of RFLP markers (DdeI, XmnI, MnlI, TaqI & HhaI), used worldwide for carrier detection, to estimate its heterozygosity in different population groups of India, and identify additional single nucleotide polymorphisms (SNPs) if necessary. A total of 8 population groups encompassing different regions of India, consisting of 107 unrelated normal females without any history of hemophilia B in the family and 13 unrelated obligate carriers were recruited in the study. Regions of F9 gene were amplified by PCR from genomic DNA of the donors followed by restriction enzyme digestion and/or sequencing as appropriate. Combined informativeness for the markers varied between 52–86% among normal females belonging to different geographical locations of India. Haplotype analysis revealed that the most prevalent haplotype lacked the restriction sites for all five RFLP markers. Screening regions of F9 gene that harbor 10 SNPs reported in dbSNP yielded only two SNPs, which increased the overall informativeness in each population group and heterozygosity in the obligate carriers for the disease from 38% to 69%. Our data show that heterozygosity of commonly used RFLP markers is remarkably variable across different regions of India. Thus prudent selection of the markers based on specific population groups including usage of additional markers is recommended for efficient carrier detection.
Apolipoprotein E in Temporal Lobe Epilepsy: A Case‐Control Study
Authors: Kumar Amit , Tripathi Manjari , Pandey Ravindra M. , Ramakrishnan Lakshmy , Srinivas M. , Luthra Kalpana
Published: June 2013

Purpose: To investigate the relationship of apolipoprotein E (apoE) genotype, plasma levels of apoE and lipids in temporal lobe epilepsy (TLE) patients in Asian Indians. Status of plasma levels of Apo E in epilepsy patients has not been reported till date.

Methods: ApoE gene polymorphism was analyzed in 58 patients with temporal lobe epilepsy (TLE) and 57 age and sex approximated controls using Polymerase chain reaction‐ restriction fragment length polymorphism (PCR‐RFLP). Levels of plasma apoE and lipids were measured using ELISA and enzymatic kits respectively.

Results: The distribution of ApoE genotype in epilepsy patients and controls was comparable. Higher levels of plasma ApoE were observed in TLE patients as compared to controls (p = 0.0001). Individuals with plasma levels of apoE > 190 mg/L were at 20 times higher odds (95%CI = 2.46–163.34, p = 0.005), while those with levels of apoE between 150–190 mg/L were at 4.9 times higher odds (95% CI = 1.85–13.9, p = 0.001), to develop TLE.

Conclusions: We have observed for the first time, high levels of plasma apoE in epilepsy patients. The findings of this case‐control study suggest that apolipoprotein E may play an important role in epilepsy.
Genome Wide Screening of CAG Trinucleotide Repeat Lengths in Breast Cancer
Authors: Jarjanazi Hamdi , Li Hong , Andrulis Irene L. , Ozcelik Hilmi
Published: June 2013

Trinucleotide repeat sequences are widely present in the human genome. The expansion of CAG repeats have been studied very extensively, and shown to be the causative mechanism of more than 40 neuromuscular and neurodegenerative diseases. In the present study, we performed a genome wide screening of CAG repeat expansions in non‐neoplastic tissues of 212 breast cancer cases and 196 healthy population controls using the Repeat Expansion Detection (RED) method. Distribution of CAG repeat lengths in cases was not significantly different from controls. However, dramatically expanded CAG repeats were detected in 2.4% (n = 5) of breast cancer cases where no repeats of similar size were detected in any of the healthy population controls. Although this trend shows only borderline significance (p = 0.06), this finding suggests a potential involvement of CAG repeat expansion in breast cancer susceptibility. These repeats may potentially affect the function of cancer predisposition genes, with a similar mechanism as in neurodegenerative and neuromuscular disorders.
The Frequency of Cytochrome P450 2E1 Polymorphisms in Black South Africans
Authors: Chelule Paul K. , Pegoraro Rosemary J. , Gqaleni Nceba , Dutton Michael F.
Published: June 2013

Polymorphisms in the promoter region of the Cytochrome P4502E1 (CYP2E1) gene reportedly modify the metabolic activity of CYP2E1 enzyme, and have been associated with increased susceptibility to squamous cell carcinoma (SCC) of the oesophagus in high prevalence areas such as China. To assess the frequency of these polymorphisms in Black South Africans, a population with a high incidence of oesophageal SCC, this study examined genomic DNA from 331 subjects for restriction fragment length polymorphisms in the CYP2E1 (RsaI and PstI digestion). The frequency of the CYP2E1 c1/c1 and c1/c3 genotypes was 95% and 5% respectively. The frequency of the CYP2E1 allele distribution was found to be markedly different between Chinese and South African populations; hence it is important to place racial differences into consideration when proposing allelic variants as genetic markers for cancer.
HPV Infections in Adolescents
Authors: Moscicki Anna-Barbara
Published: June 2013

Adolescents who are sexually active have the highest rates of prevalent and incident HPV infection rates with over 50–80% having infections within 2–3 years of initiating intercourse. These high rates reflect sexual behavior and biologic vulnerability. Most infections are transient in nature and cause no cytologic abnormality. However, a small number of adolescents will not clear the infection. Persistence of HPV is strongly linked to the development of high‐grade squamous intra‐epithelial lesions (HSIL) and invasive cancer. The HSIL detected, however, does not appear to progress rapidly to invasive cancer. Understanding the natural history of HPV in adolescents has shed light into optional treatment strategies which include watchful observation of atypical squamous cells of undetermined significance (ASCUS) and low grade (LSIL). The association between age of first intercourse and invasive cancer cannot be ignored. Consequently, initiating screening at appropriate times in this vulnerable group is essential. In addition, with the advent of the HPV vaccine, vaccination prior to the onset of sexual activity is critical since most infections occur within a short time frame post initiation.
HPV Detection Methods
Authors: Brink Antoinette A. T. P. , Snijders Peter J. F. , Meijer Chris J. L. M.
Published: June 2013

Given the causal relation between a persistent high‐risk human papillomavirus (hrHPV) infection and the development of high‐grade cervical intraepithelial neoplasia (CIN) and cervical cancer, hrHPV testing has been advocated in addition to cytology for the detection of clinically relevant cervical lesions. HrHPV testing is thought to improve cervical screening algorithms, the management of women with cytologically equivocal smears, and the management of women treated for high grade CIN. In this chapter we discuss different methods for HPV detection and genotyping and their respective applications.
The Continuum of Serous Tumors of Low Malignant Potential and Low‐Grade Serous Carcinomas of the Ovary
Authors: Wong Kwong-Kwok , Gershenson David
Published: June 2013

The role of serous tumors of low malignant potential (LMP) in the development of invasive epithelial cancer of the ovary is debatable. This review summarizes the current clinical, genetic, and genomic evidence for the existence of a continuum comprising both LMP serous tumors and low‐grade serous ovarian carcinomas.
Therapeutic Vaccination for HPV Induced Cervical Cancers
Authors: Brinkman Joeli A. , Hughes Sarah H. , Stone Pamela , Caffrey Angela S. , Muderspach Laila I. , Roman Lynda D. , Weber Jeffrey S. , Kast W. Martin
Published: June 2013

Cervical Cancer is the second leading cause of cancer–related deaths in women worldwide and is associated with Human Papillomavirus (HPV) infection, creating a unique opportunity to treat cervical cancer through anti‐viral vaccination. Although a prophylactic vaccine may be available within a year, millions of women, already infected, will continue to suffer from HPV‐related disease, emphasizing the need to develop therapeutic vaccination strategies. A majority of clinical trials examining therapeutic vaccination have shown limited efficacy due to examining patients with more advanced‐stage cancer who tend to have decreased immune function. Current trends in clinical trials with therapeutic agents examine patients with pre‐invasive lesions in order to prevent invasive cervical cancer. However, longer follow‐up is necessary to correlate immune responses to lesion regression. Meanwhile, preclinical studies in this field include further exploration of peptide or protein vaccination, and the delivery of HPV antigens in DNA‐based vaccines or in viral vectors. As long as pre‐clinical studies continue to advance, the prospect of therapeutic vaccination to treat existing lesions seem good in the near future. Positive consequences of therapeutic vaccination would include less disfiguring treatment options and fewer instances of recurrent or progressive lesions leading to a reduction in cervical cancer incidence.
Matrix Metalloproteinase Expression Patterns in Luminal A Type Breast Carcinomas
Authors: Decock J. , Hendrickx W. , Drijkoningen M. , Wildiers H. , Neven P. , Smeets A. , Paridaens R.
Published: June 2013

Objective: Aberrant expression of individual matrix metalloproteinases has been associated with poor prognosis in various human carcinomas. The current study aimed at defining an RNA expression profile of various MMPs in breast cancer and correlating their expression with clinicopathological parameters.

Methods: The RNA expression patterns of 6 MMPs (MMP2, MMP8, MMP9, MMP10, MMP11, MMP13) were determined in 25 breast carcinomas using quantitative RT‐PCR and correlated with clinicopathological parameters, including menopausal status, tumor size and grade, and lymph node involvement.

Results: We observed high MMP2 levels more frequently in premenopausal than in postmenopausal women (p = 0.02). Analysis of luminal A type invasive ductal carcinomas (19/25), revealed an even stronger association of MMP2 with menopausal status (p = 0.005). Within this subgroup, we also found a correlation between MMP11 and menopausal status (p = 0.02). No correlation was found between MMP expressions and other clinicopathological parameters. In co‐expression analyses MMP2‐MMP10 and MMP8‐MMP9 showed a weak correlation of their expression.

Conclusions: Although this is a pilot study, our findings indicate that luminal A invasive ductal carcinomas commonly express high MMP2 and MMP11 levels in premenopausal breast cancer patients and suggest a co‐regulation of MMP2‐MMP10 and MMP8‐MMP9.
Statistical Methods for Evaluating DNA Methylation as a Marker for Early Detection or Prognosis
Authors: Alonzo Todd A. , Siegmund Kimberly D.
Published: June 2013

We summarize standard and novel statistical methods for evaluating the classification accuracy of DNA methylation markers. The choice of method will depend on the type of marker studied (qualitative/quantitative), the number of markers, and the type of outcome (time‐invariant/time‐varying). A minimum of two error rates are needed for assessing marker accuracy: the true‐positive fraction and the false‐positive fraction. Measures of association that are computed from the combination of these error rates, such as the odds ratio or relative risk, are not informative about classification accuracy. We provide an example of a DNA methylation marker that is strongly associated with time to death (logrank p = 0.0003) that is not a good classifier as evaluated by the true‐positive and false‐positive fractions. Finally, we would like to emphasize the importance of study design. Markers can behave differently in different groups of individuals. It is important to know what factors may affect the accuracy of a marker and in which subpopulations the marker may be more accurate. Such an understanding is extremely important when comparing marker accuracy in two groups of subjects.
Epigenetics—DNA‐Based Mirror of our Environment?
Authors: Cooney Craig A.
Published: June 2013

Epigenetics affects health, appearance and behavior and propagates mammalian phenotypes across generations. Nutrients, drugs and behavior can all direct changes in epigenetics. In at least some cases, these directed changes are propagated across generations. This range of influences on epigenetics suggests that epigenetics is highly interactive with the environment. Changes in the environment may regularly change epigenetics and influence our future responses to the environment. The current research challenge is to understand these influences and use them to direct epigenetics toward improved health and longevity.
Overexpression of Fascin‐1 in Advanced Colorectal Adenocarcinoma: Tissue Microarray Analysis of Immunostaining Scores with Clinicopathological Parameters
Authors: Tsai Wen-Chiuan , Chao You-Chen , Sheu Lai-Fa , Chang Junn-Liang , Nieh Shin , Jin Jong-Shiaw
Published: June 2013

Objective: Fascin‐1 is an actin‐binding protein that promotes cell proliferation, adhesion and motility. We tested the hypothesis that fascin‐1 expression correlates with clinicopathological parameters of colorectal adenocarcinomas.

Methods: Immunohistochemical analysis of fascin‐1 was performed in tissue microarrays of 91 surgical specimens, including 32 well, 33 moderately, and 26 poorly differentiated colorectal adenocarcinomas; and in 22 specimens from colorectal adenomas with dysplasia.

Results: Scattered fascin‐1 expression was demonstrated in 9 control specimens of normal colonic glandular epithelia. Higher fascin‐1 immunostaining scores were significantly associated with advanced dysplasia in colorectal adenomas (mild 4.2 ± 1.3, moderate 13.5 ± 5.3, and severe 22.5 ± 6.7) and high‐grade histopathological differentiation of colorectal adenocarcinomas (grade I 88.6 ± 9, grade II 101 ± 11, and grade III 144 ± 13). Higher immunostaining scores of fascin‐1 were also significantly associated with advanced T stage (T1: 42 ± 10, T2: 60 ± 12, T3: 108 ± 12, and T4: 142 ± 15). Higher fascin‐1 scores were related with more advanced M and N stages of colorectal carcinomas, but not significant correlation.

Conclusions: Higher expression of fascin‐1 correlates significantly with tumor grades and TNM stages in colorectal adenocarcinomas and also with levels of dysplastic change in colorectal adenomas.
Human Papillomaviruses and Non‐Melanoma Skin Cancer: Basic Virology and Clinical Manifestations
Authors: Nindl Ingo , Gottschling Marc , Stockfleth Eggert
Published: June 2013

Human papillomaviruses (HPV) infect cutaneous and mucosal epithelia and induce benign and malignant lesions. Non‐melanoma skin cancer (NMSC), encompassing basal cell carcinoma and squamous cell carcinoma (SCC), is the most frequent cancer in the Caucasian population, and the incidence has increased dramatically worldwide. Ultraviolet (UV) radiation is a major risk factor for NMSC, and cutaneous HPV is also considered to play an active role during the pathogenesis of these cancers. The first evidence for the involvement of HPV in NMSC was reported in patients with Epidermodysplasia verruciformis (EV). HPV types detected in skin tumours of these patients are referred to as EV/cutaneous HPV types belonging to the beta‐ and gamma‐papillomaviruses (PV). Epidemiological studies have shown a higher risk of several EV/cutaneous HPV types for NMSC. Furthermore, in vitro and animal models show transforming properties of some PV types. The anti‐apoptotic activities, and the delay of DNA repair mechanism caused by some EV/cutaneous HPV E6 proteins in response to UV‐induced mutations, may lead to the persistence of DNA‐damaged keratinocytes. Thus, specific EV/cutaneous HPV types as co‐factors in association with UV‐radiation and the immune system seem to be involved in the early pathogenesis of cutaneous SCC.
Low Plasma Levels of FGF‐2 and PDGF‐BB Are Associated with Cardiovascular Events in Type II Diabetes Mellitus (Diabetes Heart Study)
Authors: Yeboah Joseph , Sane David C. , Crouse John R. , Herrington David M. , Bowden Donald W.
Published: June 2013

Objective: We tested associations of the growth factors VEGF, FGF‐2, HGF and PDGF‐BB with coronary artery calcium scores and cardiovascular events (CVD) in type 2 diabetes mellitus (T2DM).

Methods: A cross‐sectional study selected 40 frequency matched (by age, gender and race) subjects with T2DM from the first (0–111) and the third (> 1400) coronary artery calcium (CAC) score tertiles in the Diabetes Heart Study (DHS), in which 36 were with and 41 were without history of CVD events. Plasma levels of VEGF, FGF‐2, HGF and PDGF‐BB were measured in all subjects.

Results: None of the growth factors was significantly different between the first and third CAC score tertiles. Mean plasma FGF‐2 and PDGF‐BB levels were significantly higher in the group without prior CVD events compared with the group with prior CVD events [mean(95%CI): 219.20 (194.42–247.15) vs. 152.93 (135.64–172.43) pg/ml, p = 0.03] and [mean(95%CI): 106.70 (89.12–127.74) vs. 61.56 (50.91–74.44) pg/ml, p = 0.03], respectively. Subgroup analysis in the first CAC tertile showed a significantly higher PDGF‐BB levels in those without compared with those with CVD events [mean (95%CI): 208.36 (190.57–228.15) vs. 102.93 (80.64–125.21) pg/ml, p = 0.004].

Conclusion: Plasma growth factor levels were not significantly different between the extremes of CAC scores in T2DM. However, low plasma levels of PDGF‐BB and FGF‐2 are associated with prior cardiovascular events in T2DM. Studies are needed to confirm our results and also to establish temporality of this association.
The Role of DNA Methylation in the Development and Progression of Lung Adenocarcinoma
Authors: Kerr Keith M. , Galler Janice S. , Hagen Jeffrey A. , Laird Peter W. , Laird-Offringa Ite A.
Published: June 2013

Lung cancer, caused by smoking in ∼87% of cases, is the leading cause of cancer death in the United States and Western Europe. Adenocarcinoma is now the most common type of lung cancer in men and women in the United States, and the histological subtype most frequently seen in never‐smokers and former smokers. The increasing frequency of adenocarcinoma, which occurs more peripherally in the lung, is thought to be at least partially related to modifications in cigarette manufacturing that have led to a change in the depth of smoke inhalation. The rising incidence of lung adenocarcinoma and its lethal nature underline the importance of understanding the development and progression of this disease. Alterations in DNA methylation are recognized as key epigenetic changes in cancer, contributing to chromosomal instability through global hypomethylation, and aberrant gene expression through alterations in the methylation levels at promoter CpG islands. The identification of sequential changes in DNA methylation during progression and metastasis of lung adenocarcinoma, and the elucidation of their interplay with genetic changes, will broaden our molecular understanding of this disease, providing insights that may be applicable to the development of targeted drugs, as well as powerful markers for early detection and patient classification.
Analysis of Crohn’s Disease‐Related CARD15 Polymorphisms in Spanish Patients with Idiopathic Uveitis
Authors: Rodríguez-Pérez N. , Aguinaga-Barrilero A. , Gorroño-Echebarría Marina B. , Pérez-Blas Mercedes , Martín-Villa J. M.
Published: June 2013

We wished to analyse the frequency of Crohn’s disease‐linked CARD15 polymorphisms (P268S, R702W, G908R and 1007fs) in a group of Spanish patients with idiopathic uveitis. To this aim, DNA samples were obtained from 111 unrelated patients. P268S, R702W and G908R polymorphisms were detected using TaqMan Genotyping kits (Applied Biosystems), and the 1007fs variation by direct DNA sequencing. Control group consisted of 105 healthy subjects.

None of the polymorphisms studied revealed a significant increase in the groups of patients, when compared to the control group. Thus, P268S is found in 50% of patients (gene frequency 0.284) vs 44% of control individuals (gene frequency 0.245); R702W in 7% of patients (0.036) vs 7% (0.033); G908R in 2% of patients (0.009) vs 4% (0.019) and, finally, 1007fs in 2% of uveitis patients (0.008) vs 4% (0.021). Moreover, DNA sequencing has allowed us to define two new intronic polymorphisms in phase, in the 5′ and 3′ boundaries of the exon 11 (GenBank accession number #DQ 869189).

Altogether, our results suggest that the Crohn’s disease‐linked CARD15 polymorphisms do not seem to predispose to idiopathic uveitis in the Spanish population.
Stem Cell Biomarkers in Chronic Myeloid Leukemia
Authors: Jiang Xiaoyan , Zhao Yun , Forrest Donna , Smith Clayton , Eaves Allen , Eaves Connie
Published: June 2013

Chronic myeloid leukemia (CML) is a clonal multi‐step myeloproliferative disease that is initially produced and ultimately sustained by a rare subpopulation of BCR-ABL+ cells with multi‐lineage stem cell properties. These BCR-ABL+ CML stem cells are phenotypically similar to normal hematopoietic stem cells which are also maintained throughout the course of the disease at varying levels in different patients. Defining the unique properties of the leukemic stem cells that produce the chronic phase of CML has therefore had to rely heavily on access to samples from rare patients in which the stem cell compartment is dominated by leukemic elements. Here we review past and ongoing approaches using such samples to identify biologically and clinically relevant biomarkers of BCR-ABL+ stem cells that explain their unusual biology and that may help to design, or at least predict, improved treatment responses in CML patients. These studies are of particular interest in light of recent evidence that chronic phase CML stem cells are not only innately resistant to imatinib mesylate and other drugs that target the BCR‐ABL oncoprotein, but are also genetically unstable.
The C677T Polymorphism in MTHFR Is Not Associated with Migraine in Portugal
Authors: Ferro Anabela , Castro Maria-José , Lemos Carolina , Santos Mónica , Sousa Alda , Pereira-Monteiro José , Sequeiros Jorge , Maciel Patrícia
Published: June 2013

Migraine is a debilitating disorder affecting a large proportion of the population. The effect of methylenetrathydrofolate reductase (GeneID: 4524) polymorphisms in migraine etiology and development has been a theme of great interest. Several populations were evaluated with contradictory results. In this case‐control study, we investigated the effect of the C677T polymorphism in MTHFR, as a genetic risk factor for migraine, in the Portuguese population. We observed that, overall, there was no significant difference in the frequencies of MTHFR C677T genotypes or of the T‐allele among the Portuguese migraineurs when compared to controls. There was also no association of migraine with aura with MTHFR genotypes or with the T‐allele, in contrast with previous studies. Regarding the risk of the T‐allele homozygote carriers, there was an equal probability to develop migraine with aura over migraine without aura in our patients. Thus, we conclude that the C677T MTHFR polymorphism, responsible for a reduction of the MTHFR activity in folate metabolism, is not a major genetic susceptibility factor for migraine in the Portuguese population.
Erythrocyte Aggregation As an Early Biomarker in Patients with Asymptomatic Carotid Stenosis
Authors: Assayag Einor Ben , Bova Irena , Kesler Anat , Berliner Shlomo , Shapira Itzhak , Bornstein Natan M.
Published: June 2013

Background: Atherosclerosis is a chronic inflammatory disease.

Design: We have evaluated the degree of erythrocyte aggregation (EA) as a microinflammatory biomarker in a cohort of hospital‐based, neurologically asymptomatic outpatients.

Methods: The degree of EA and carotid artery stenosis was evaluated in 510 individuals by using a simple slide test and image analysis.

Results: Four hundred and sixteen individuals had minimal carotid stenosis (< 30%); 47 had mild to moderate stenosis (30–69%)
and 47 had severe stenosis (>70%). A significant correlation was noted between the degree of carotid stenosis and the erythrocyte sedimentation rate (ESR), white blood cell count (WBCC) and fibrinogen (r=0.160, p=0.005;r=0.191, p=0.001 andr=0.126, p=0.026, respectively). The significant correlation was noted between the degree of carotid stenosis and EA (r=0.209, p < 0.001). The subjects with severe stenosis differed significantly from the other groups in their ESR, WBCC and EA.
High sensitivity C‐reactive protein (hs‐CRP) concentrations did not discriminate between the presence and absence of significant
carotid atherosclerotic disease.

Conclusions: Inflammatory biomarkers such as ESR and the EA test are more sensitive than hs‐CRP to the presence of a significant atherosclerotic carotid burden. These biomarkers might aid in the detection and quantification of microinflammation in individuals with carotid atherosclerosis.
Association of Interleukin‐10 Gene Promoter Polymorphisms in Saudi Patients with Vitiligo
Authors: Abanmi Abdullah , Al Harthi Fahad , Zouman Abdulrahman , Kudwah Aida , Jamal Mohammed Al , Arfin Misbahul , Tariq Mohammad
Published: June 2013

The promoter region of human Interleukin −10 gene is highly polymorphic and has been associated with numerous autoimmune diseases. Recent studies have linked vitiligo with defective autoimmune system. This study is aimed to explore a possible association between IL‐10 gene polymorphism and vitiligo in Saudi population. This case control study consisted of 184 Saudi subjects including 83 vitiligo patients (40 males, 43 females mean age 27.85 ± 12.43 years) and 101 matched controls. Genomic DNA was extracted from the blood samples of healthy controls and Vitiligo patients visiting out patient clinic of Department of Dermatology, Riyadh Armed Forces Hospital, using QIA ampR DNA mini kit (Qiagen CA, USA). Interleukin‐10 gene was amplified by polymerase chain reaction (PCR) using Arms primers to detect any polymorphism involved at positions −592, −819 and −1082.

The frequencies of GG genotype at −1082, and CC genotype at positions −592 and 819 were significantly higher in vitiligo patients compared to healthy subjects suggesting that GG and CC genotypes might be susceptible to vitiligo in Saudis. On the other hand genotypes −1082 GA, −819 CT, and −592 CA of IL‐10 were more prevalent in healthy controls suggesting protective effects of GA, CT and CA genotypes against vitiligo. This study indicates that the IL‐10 gene may play a significant role in the etiology of vitiligo among Saudis.
Fibronectin Pattern in Benign Hyperplasia and Cancer of the Prostate
Authors: Janković Miroslava M. , Kosanović Maja M.
Published: June 2013

Fibronectin (FN) is a multifunctional glycoprotein involved in cell‐matrix interactions. It exhibits a complex pattern of forms differing in respect to aminoacid and oligosaccharide composition. In this study we examined glycobiochemical and functional properties of the FN in benign prostatic hyperplasia (BPH) and prostatic cancer (PCa), attempting to resolve disease‐related differences. Two BPH sera pools and three PCa sera pools were used as the FN source. The affinity‐purified molecule was characterized by SDS‐PAGE, immuno‐ and lectin blot, lectin‐affinity chromatography and adhesion assay. BPH FN existed as intact molecule, giving the main immunoreactive band at 220 kDa. In contrast, PCa FN comprised three main immunoreactive fragments of 140, 110 and 90 kDa. As for glycosylation the ratio of altogether lectin‐reactive PCa FN was different from that of BPH FN manifested as a decrease of Con A‐ and an increase of LCA‐reactive moieties. Fibroblasts adhered to both FN preparations in a concentration dependent manner, but with a significantly lower efficiency to PCa FN. The results obtained showing distinct structural characteristics of PCa FN compared to BPH FN could be important for modulation of its ligand and recognition properties expressed as gain or loss of functions or as specific markers of its origin.
Stability and Reliability of Plasma Level of Lipid Biomarkers and Their Correlation with Dietary Fat Intake
Authors: Lee Sang-Ah , Wen Wanqing , Xiang Yong-Bing , Fazio Sergio , Linton MacRae F. , Cai Qiuyin , Liu Dake , Zheng Wei , Shu Xiao-Ou
Published: June 2013

The reliability and stability of plasma lipid biomarkers and their association with dietary fat intake were evaluated among 48 subjects who were randomly chosen from the participants of a validation study of the population‐based cohort, the Shanghai Men′s Health Study (SMHS). Four spot blood samples, one taken each season, were measured for total cholesterol, triglyceride, HDL‐cholesterol, and LDL‐cholesterol levels. The reliability and stability of these measurements were assessed by intraclass correlation coefficients (ICC) and by the correlations between a randomly chosen measurement with the mean of measurements across seasons using a bootstrap approach. The median levels for total cholesterol, triglycerides, HDL‐cholesterol, and LDL‐cholesterol were 177.5, 164.5, 41.0, and 102.5 (mg/dl), respectively. The ICCs of the biomarkers ranged from 0.58 (LDL‐cholesterol) to 0.83 (HDL‐cholesterol). The correlation between randomly chosen spot measurements and the mean measurement were 0.91, 0.86, 0.93, and 0.83 for total cholesterol, triglycerides, HDL‐cholesterol, and LDL‐cholesterol, respectively. The correlations of lipid biomarkers with dietary fat intake and other lifestyle factors were comparable to other previous reports. In conclusion, this study suggests that measurements of lipid biomarkers from a single spot blood sample are a good representation of the average blood levels of these biomarkers in the study population and could be a useful tool for epidemiological studies.
The oxidation ratio of LDL: A Predictor for Coronary Artery Disease
Authors: Huang Huiling , Mai Weiyi , Liu Dan , Hao Yuantao , Tao Jun , Dong Yugang
Published: June 2013

Objective: Oxidized LDL cholesterol (ox‐LDL‐C) is considered to be a key factor of initiating and accelerating atherosclerosis (AS). The purpose of this study is to elucidate the sensitivity and specificity of ox‐LDL and oxidation ratio of LDL in the diagnosis of coronary artery disease (CAD). For the first time, we investigated the ratio of ox‐LDL to ALB(ox‐LDL/ALB).

Methods and results: Blood ox‐LDL, total cholesterol (TC), high density lipoprotein cholesterol (HDL‐C), low density lipoprotein cholesterol (LDL‐C), triglyceride (TG) and albumin (ALB) were measured in patients with acute myocardial infarction (AMI, n = 80), unstable angina pectoris (UAP, n = 80), stable angina pectoris (SAP, n = 80), normal control (n = 60), and dyslipidemia control (n = 60). Ox‐LDL was measured by competitive ELISA. The level of ox‐LDL and oxidation ratio of LDL(ox‐LDL/TC, ox‐LDL/HDL‐C, ox‐LDL/ LDL‐C and ox‐LDL/ALB) were significantly higher in each diseased group than controls (P < 0.001). In CAD group, ox‐LDL and oxidation ratio of LDL in subjects complicated with hypertension (HT) and/or diabetes mellitus (DM) increased further (P < 0.001). Ox‐LDL/ALB in the AMI group was 7 times higher than normal control group (0.068 ± 0.017 vs 0.009 ± 0.007, P < 0.001). The area under the curve (AUC) of receiver operating characteristic curve (ROC curve) is a criterium to evaluate the accuracy of diagnosing a disease. The AUC of ROC curve of ox‐LDL/TC, ox‐LDL/HDL‐C, ox‐LDL, ox‐LDL/ALB and ox‐LDL/ LDL‐C for diagnosing CAD were 0.975, 0.975, 0.966, 0.966, 0.957 respectively (P < 0.001). When ox‐LDL/TC = 0.175, the sensitivity and specificity of diagnosing CAD were 0.917 and 0.925, which were almost equal to each other, indicating that the rates of missed diagnosis and misdiagnosis for CAD were the lowest.

Conclusions: The level of ox‐LDL and the ratio of ox‐LDL/TC, ox‐LDL/LDL‐C, ox‐LDL/HDL‐C and ox‐LDL/ALB are better biomarkers than TC, TG, HDL‐C and LDL‐C for discriminating between patients with coronary artery disease and healthy subjects. And patients who have a high ratio of ox‐LDL /TC may have a higher risk for CAD.
Lung Cancer Stem Cells
Authors: Pine Sharon R. , Marshall Blair , Varticovski Lyuba
Published: June 2013

Lung cancer remains a major cause of cancer‐related lethality because of high incidence and recurrence in spite of significant advances in staging and therapies. Recent data indicates that stem cells situated throughout the airways may initiate cancer formation. These putative stem cells maintain protumorigenic characteristics including high proliferative capacity, multipotent differentiation, drug resistance and long lifespan relative to other cells. Stem cell signaling and differentiation pathways are maintained within distinct cancer types, and destabilization of this machinery may participate in maintenance of cancer stem cells. Characterization of lung cancer stem cells is an area of active research and is critical for developing novel therapies. This review summarizes the current knowledge on stem cell signaling pathways and cell markers used to identify the lung cancer stem cells.
CYP17 MspA1 Polymorphism and Age at Menarche: A Meta‐Analysis
Authors: Pei Yu-Fang , Zhang Lei , Deng Hong-Wen , Dvornyk Volodymyr
Published: June 2013

Objective: Literature data on the effects of CYP17 MspA1 polymorphism on age at menarche (AAM) are inconsistent. To reexamine this controversy, we performed a meta‐analysis.

Study design: In total 16 studies containing more than 11000 individuals of various ethnicities were selected for the analyses. For 11 case‐control studies, odds ratio (OR) was employed to evaluate the risk of late AAM for each study, using homozygote at the wild‐type allele as a control group. For the 5 studies with continuous outcomes, the effect size was estimated using the Hedges’ adjusted g, which is calculated based on the standardized mean difference between groups of subjects with early and late AAM.

Results: We did not find evidence for association of the MspA1 polymorphism with AAM in the combined case‐control sample with mixed ethnic background (OR = 1.03, 95% CI: 0.90–1.18, P = 0.66), in the monoethnic case‐control sample of Caucasian females (OR = 1.09, 95% CI: 0.99–1.20, P = 0.08) and in the combined sample with continuous traits (Hedges’ g = 0.33 and −0.041, 95% CI: −0.14–0.80 and −0.18–0.10, P values 0.17 and 0.56 for the pooled population sample and monoethnic sample of Caucasian females, respectively).

Conclusion: Our study showed that CYP17 MspA1 polymorphism was not a significant independent risk factor of AAM. Further studies are needed to clarify the effects of the interaction between this gene and other genetic and/or environment factors on AAM.
The impact of Folate Pathway Polymorphisms Combined to Nutritional Deficiency As a Maternal Predisposition Factor for Down Syndrome
Authors: Santos-Rebouças C. B. , Corrêa J. C. , Bonomo A. , Fintelman-Rodrigues N. , Moura K. C. V. , Rodrigues C. S. C. , Santos J. M. , Pimentel M. M. G.
Published: June 2013

Polymorphisms in genes encoding folate metabolizing enzymes have been linked to an increased risk of maternal chromosomal nondisjunction in several populations. With the purpose of evaluating this relationship, we compared the frequencies of 677C>T and 1298A>C polymorphisms in the methylenetetrahydrofolate reductase gene (MTHFR) and 66A>G in the methionine synthase reductase gene (MTRR) between 103 young mothers of Down syndrome (DS) individuals and 108 control mothers, whose offspring was karyotypically normal, correlating it with an estimative of folate and – related micronutrients levels intake. Maternal and paternal transmission frequencies of MTHFR 677T allele were also examined to access potential parent‐of‐origin effects. PCR‐RFLP for genomic DNA was accomplished and allele/genotype frequencies differences were determined using the x2 test, whereas pattern of transmission of the MTHFR 677 allele was analyzed by transmission disequilibrium test. None of the polymorphisms seemed to be more frequent in case mothers than in controls, either individually or combined. The estimative of nutritional intake revealed that folate consumption median was inadequate in both groups, whereas methionine and zinc consumption medians were significantly greater in control mothers. It suggests that such interaction between genetic profile and environment could predispose this sub group of women to have a DS child. Additional studies focusing the interaction between nutritional intakes, biochemical data and folate pathway polymorphisms are needed to confirm the present results. The possibility of neutralize the biochemical negative effects of folate‐related polymorphisms through oral supplementation could provide new targets for DS prevention.
Glycosylation of Serum Proteins in Inflammatory Diseases
Authors: Gornik Olga , Lauc Gordan
Published: June 2013

Inflammatory diseases are accompanied by numerous changes at the site of inflammation as well as many systemic physiological and biochemical changes. In the past two decades more and more attention is being paid to changes in glycosylation and in this review we describe some of the changes found on main serum proteins (α1‐acid glycoprotein, immunoglobulin G, immunoglobulin A, transferrin, haptoglobin, α2‐macroglobulin, C‐reactive protein, and others). Molecular background and physiological importance of most of these changes are yet to be discovered, but it is evident that glycosylation plays an important role in the inflammatory response. Maybe the greatest value of these changes currently lays in their potential diagnostic and prognostic usage, either in combination with current diagnostic markers or on their own. However, determining glycan structures is still technically too complex for most clinical laboratories and further efforts have to be made to develop simple analytical tools to study changes in glycosylation.
Glycomic Approach for Potential Biomarkers on Prostate Cancer: Profiling of N‐Linked Glycans in Human Sera and pRNS Cell Lines
Authors: de Leoz Maria Lorna A. , An Hyun Joo , Kronewitter Scott , Kim Jaehan , Beecroft Sean , Vinall Ruth , Miyamoto Suzanne , de Vere White Ralph , Lam Kit S. , Lebrilla Carlito
Published: June 2013

Prostate cancer is a leading cause of cancer death among men. Currently available screening test measures prostate‐specific antigen (PSA) to detect prostate cancer. However, this test produces false positive values that often lead to negative biopsies. Therefore, a more reliable diagnostic tool is needed. Glycans in serum are of particular interest as around half of all proteins are glycosylated. In this study, N‐linked glycans were enzymatically released by PNGase F from prostate epithelial cell lines (pRNS) expressing wild type or mutant androgen receptors and a small set of human serum samples. Released glycans were purified and partitioned into neutral and acidic components by solid phase extraction (SPE) using graphitized carbon cartridges. The SPE fractions were analyzed by matrix‐assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry (MALDI FT‐ICR MS). Significant changes in some high‐mannose and fucosylated biantennary complex N‐linked glycans were observed in the serum of prostate cancer patients.
Sensitivity and Frequencies of Dystrophin Gene Mutations in Thai DMD/BMD Patients As Detected by Multiplex PCR
Authors: Sura Thanyachai , Eu-ahsunthornwattana Jakris , Pingsuthiwong Sarinee , Busabaratana Manisa
Published: June 2013

Background: Duchenne muscular dystrophy (DMD), a lethal X‐linked disease affecting 1 in 3500 male births, and its more benign variant, Becker muscular dystrophy (BMD), are caused by mutations in the dystrophin gene. Because of its large size, analysing the whole gene is impractical. Methods have been developed to detect the commonest mutations i.e. the deletions of the exons. Although these tests are highly specific, their sensitivity is inherently limited by the prevalence of deletions, which differs among different populations.

Methods: We reviewed our database for the detection of Dystrophin gene mutation by means of 31‐exon multiplex PCR in Thai males, diagnosed clinically and biochemically with DMD or BMD from July 1994 to November 2006. One index patient was chosen from each family for statistical analysis. The overall sensitivity of the test, the number of fragment deleted, and the deletion frequency of each fragment were calculated, along with their 95% confidence intervals (C.I.).

Results: We found deletions in 99 out of the 202 index patients (49%; Bayesian 95% C.I. = 42%–56%). 51% of these had deletion in only one of the 31 exons tested, while the patient with the most extensive deletions had 14 exons deleted. The mean number of deleted exons were 2.84 (BCa bootstrap 95% C.I. = 2.37–3.48), or 5.02 (3.81–6.85) if all the untested exons adjacent to the confirmed deleted exons were assumed to be deleted. The region spanning exons 44‐52 was the most frequently deleted. These were similar to those reported in the Japanese.

Conclusion: The multiplex PCR detected deletions only in about half of the Thai patients. The diseases therefore should not be excluded solely on the negative result if DMD/BMD is strongly suspected.
In Situ Nuclear Morphology Measurements Using Light Scattering as Biomarkers of Neoplastic Change in Animal Models of Carcinogenesis
Authors: Wax Adam , Pyhtila John W.
Published: June 2013

Light scattering spectroscopy measurements can be used to determine the structure of tissue samples. Through refined data acquisition and signal processing techniques, quantitative nuclear morphology measurements may be obtained from light scattering data. These data have been used primarily as a biomarker of neoplastic change in a wide range of settings. Here, we review the application of light scattering to assessing the health status of tissues drawn from animal models of carcinogenesis, in particular, the rat esophagus and the golden Syrian hamster trachea carcinogenesis models. In addition, we present results from ex vivo human tissues to demonstrate the relevance of the use of animal models which are excellent surrogates for several human cancers. These models provide the opportunity to develop biomarkers and test chemopreventive and therapy strategies before application in humans.
Inflammation, Adenoma and Cancer: Objective Classification of Colon Biopsy Specimens with Gene Expression Signature
Authors: Galamb Orsolya , Györffy Balázs , Sipos Ferenc , Spisák Sándor , Németh Anna Mária , Miheller Pál , Tulassay Zsolt , Dinya Elek , Molnár Béla
Published: June 2013

Gene expression analysis of colon biopsies using high‐density oligonucleotide microarrays can contribute to the understanding of local pathophysiological alterations and to functional classification of adenoma (15 samples), colorectal carcinomas (CRC) (15) and inflammatory bowel diseases (IBD) (14). Total RNA was extracted, amplified and biotinylated from frozen colonic biopsies. Genome‐wide gene expression profile was evaluated by HGU133plus2 microarrays and verified by RT‐PCR. We applied two independent methods for data normalization and used PAM for feature selection. Leave one‐out stepwise discriminant analysis was performed. Top validated genes included collagenIVα1, lipocalin‐2, calumenin, aquaporin‐8 genes in CRC; CD44, met proto‐oncogene, chemokine ligand‐12, ADAM‐like decysin‐1 and ATP‐binding casette‐A8 genes in adenoma; and lipocalin‐2, ubiquitin D and IFITM2 genes in IBD. Best differentiating markers between Ulcerative colitis and Crohn′s disease were cyclin‐G2; tripartite motif‐containing‐31; TNFR shedding aminopeptidase regulator‐1 and AMICA. The discriminant analysis was able to classify the samples in overall 96.2% using 7 discriminatory genes (indoleamine‐pyrrole‐2,3‐dioxygenase, ectodermal‐neural cortex, TIMP3, fucosyltransferase‐8, collectin sub‐family member 12, carboxypeptidase D, and transglutaminase‐2). Using routine biopsy samples we successfully performed whole genomic microarray analysis to identify discriminative signatures. Our results provide further insight into the pathophysiological background of colonic diseases. The results set up data warehouse which can be mined further.