bluuebunny/crossref_metadata_2025 huggingface dataset:
Circulating Cell‐Free DNA in Plasma of Locally Advanced Rectal Cancer Patients Undergoing Preoperative Chemoradiation: A Potential Diagnostic Tool for Therapy Monitoring
Authors: Zitt Matthias , Müller Hannes M. , Rochel Marina , Schwendinger Verena , Zitt Marion , Goebel Georg , DeVries Alexander , Margreiter Raimund , Oberwalder Michael , Zeillinger Robert , Öfner Dietmar
Published: June 2013

Circulating cell‐free DNA opens up an interesting field for therapy monitoring, in particular during multimodal therapy protocols. The objective of this proof of principle study was to evaluate whether the amount of circulating plasma DNA has the potential to serve as a marker for therapy monitoring during the treatment course of locally advanced rectal cancer patients. We especially focused on kinetics of circulating DNA to assess whether variances in kinetics have the potential to discriminate between therapy responders and nonresponders.

The amount of circulating DNA in plasma of rectal cancer patients undergoing preoperative chemoradiation was determined using real‐time PCR before chemoradiation, after the end of chemoradiation and at the end of treatment. The study population was divided into responders (ypT0‐T2 stage) and nonresponders (ypT3‐T4 stage). Both groups showed comparable median plasma DNA values before and after the end of chemoradiation. At the end of treatment responders showed a further decrease in circulating DNA, whereas in nonresponders the circulating DNA manifestly increased (P = 0.006).

This study demonstrates that circulating DNA in plasma of rectal cancer patients undergoing preoperative chemoradiation might serve as a surrogate marker to discriminate between responders and nonresponders. Therefore, we hypothesize that quantification of plasma DNA could be of use as an easily accessible tool for therapy monitoring in these patients.
Polymorphisms of Toll Like Receptors in the Genetics of Severe RSV Associated Diseases
Authors: Mailaparambil Beena , Krueger Marcus , Heinze Jessica , Forster Johannes , Heinzmann Andrea
Published: June 2013

Toll like receptors (TLRs) are an essential part of the innate immune response. So far, ten different TLRs were identified in humans. They recognize a wide range of microbial and viral pathogens. Infection by respiratory syncytial virus (RSV) is still a major health problem, about 2% of all children are hospitalised due to RSV bronchiolitis during their first 2 years of live. TLR4 has already been described in association with RSV associated diseases by us and others.

Thus we were interested whether other TLRs are also involved in the genetics of severe RSV infection. We genotyped 19 polymorphisms in the autosomal TLRs, these are TLR1, 2, 3, 5, 6, 9 and 10. Association analyses by the Armitage′s Trend test revealed weak association of one TLR9 promoter polymorphism with RSV infection (p = 0.013). In addition, association was found with TLR10 haplotypes (p = 0.024).

We conclude from our data – that – although we can not rule out a minor involvement of TLR9 polymorphism and TLR10 haplotypes – TLRs other than TLR4 do not play a major role in the genetics of severe RSV associated diseases. Future studies should focus on additional genes of the innate immune response.
Detection of Membrane Fluidity in Submitochondrial Particles of Platelets and Erythrocyte Membranes from Mexican Patients with Alzheimer Disease by Intramolecular Excimer Formation of 1,3 Dipyrenylpropane
Authors: Ortiz G.G. , Pacheco-Moisés F. , El Hafidi M. , Jiménez-Delgado A. , Macías-Islas M. A. , Corral S. A. Rosales , de la Rosa A. Célis , Sánchez-González V. J. , Arias-Merino E. D. , Velázquez-Brizuela I. E.
Published: June 2013

It has been suggested that mitochondrial dysfunction and defects in membrane structure could be implied in AD pathogenesis. The aim of the present work was the study of membrane fluidity in submitochondrial platelet particles and erythrocyte membranes from Mexican patients. Blood samples were obtained from 30 patients with Alzheimer disease and 30 aged‐matched control subjects. Membrane fluidity determinations were done using a very low concentration of the fluorescent dipyrenylpropane probe incorporated in both types of membranes. This probe is able to give excimer and monomer fluorescence, therefore it can be used to monitor fluidity changes in biological membranes.

The data obtained showed that in submitochondrial particles from AD patients, the excimer to monomer fluorescent intensity ratio was lower (0.231 ± 0.008) than aged‐matched control subjects (0.363 ± 0.014). Therefore, membrane fluidity was lower in AD samples. On the other hand, we found similar membrane fluidity in erythrocytes from AD patients and aged‐matched controls: the fluorescent intensity ratios were 0.312 ± 0.03 and 0.305 ± 0.033, respectively. In addition, lipid peroxidation in submitochondrial particles and erythrocyte membranes was higher in AD samples than in aged‐matched controls. These data suggest that submitochondrial platelet particles are more sensitive to oxidative stress than erythrocyte membranes.
Mast Cells, TGF‐β1 and α‐SMA Expression in IgA Nephropathy
Authors: Silva G. E. B. , Costa R. S. , Ravinal R. C. , dos Reis M. A. , Dantas M. , Coimbra T. M.
Published: June 2013

IgA nephropathy (IgAN) is a kidney disease with a varying renal prognosis. Recently, many studies have demonstrated that renal α‐smooth muscle actin (α‐SMA) and transforming growth factor (TGF‐β1) expression, as well interstitial mast cell infiltrates could represent a prognostic marker in several renal diseases. The aim of our study was to analyze the prognostic value of mast cell, TGF‐β1 and α‐SMA expression in IgAN. A survey of the medical records and renal biopsy reports of 62 patients with a diagnosis of IgAN followed‐up from 1987 to 2003 was performed. The mean follow‐up time was 74.7 ± 50.0 months. The immunohistochemical studies were performed using a monoclonal antibody anti‐human mast cell tryptase, a polyclonal antibody anti‐human TGF‐β1, and a monoclonal antibody anti‐human α‐SMA. An unfavorable clinical course of IgAN was related to interstitial mast cell infiltrates and α‐SMA expression in the tubulointerstitial area. Expression of glomerular TGF‐β1 and α‐SMA, and interstitial TGF‐β1 is not correlated with clinical course in IgAN. In conclusion, the increased number of mast cells and higher α‐SMA expression in the tubulointerstitial area may be predictive factors for the poor prognosis of patients with IgAN.
Candidate Gene Discovery Procedure after Follow‐Up Confirmatory Analyses of Candidate Regions of Interests for Alzheimer’s Disease in the NIMH Sibling Dataset
Authors: Baye Tesfaye M. , Perry Rodney T. , Wiener Howard W. , Chen Zuomin , Harrell Lindy E. , Go Rodney C. P.
Published: June 2013

The objective of this research was to develop a procedure to identify candidate genes under linkage peaks confirmed in a follow‐up of candidate regions of interests (CRIs) identified in our original genome scan in the NIMH Alzheimer’s diseases (AD) Initiative families (Blacker et al. [1]). There were six CRIs identified that met the threshold of multipoint lod score (MLS) of ≥ 2.0 from the original scan. The most significant peak (MLS = 7.7) was at 19q13, which was attributed to APOE. The remaining CRIs with ‘suggestive’ evidence for linkage were identified at 9q22, 6q27, 14q22, 11q25, and 3p26. We have followed up and narrowed the 9q22 CRI signal using simple tandem repeat (STR) markers (Perry et al. [2]). In this confirmatory project, we have followed up the 6q27, 14q22, 11q25, and 3p26 CRIs with a total of 24 additional flanking STRs, reducing the mean interval marker distance (MID) in each CRI, and substantially increase in the information content (IC). The linkage signals at 6q27, 14q22 and 11q25 remain ‘suggestive’, indicating that these CRIs are promising and worthy of detailed fine mapping and assessment of candidate genes associated with AD.

We have developed a bioinformatics approach for identifying candidate genes in these confirmed regions based on the Gene Ontology terms that are annotated and enriched among the systematic meta‐analyzed genes, confirmed by at least three case‐control samples, and cataloged in the “AlzGene database” as potential Alzheimer disease susceptibility genes (http://www.alzgene.org).
Analysis of Highly Conserved Regions of the 3’UTR of MECP2 Gene in Patients with Clinical Diagnosis of Rett Syndrome and Other Disorders Associated with Mental Retardation
Authors: Santos Mónica , Yan Jin , Temudo Teresa , Oliveira Guiomar , Vieira José Pedro , Fen Jinong , Sommer Steve , Maciel Patrícia
Published: June 2013

In this work we explored the role of the 3’UTR of the MECP2 gene in patients with clinical diagnosis of RTT and mental retardation; focusing on regions of the 3’UTR with almost 100% conservation at the nucleotide level among mouse and human. By mutation scanning (DOVAM‐S technique) the MECP2 3’UTR of a total of 66 affected females were studied. Five 3’UTR variants in the MECP2 were found (c.1461+9G>A, c.1461+98insA, c.2595G>A, c.9961C>G and c.9964delC) in our group of patients. None of the variants found is located in putative protein‐binding sites nor predicted to have a pathogenic role. Our data suggest that mutations in this region do not account for a large proportion of the RTT cases without a genetic explanation.
CYP3A5*3, CYP3A4*1B and MDR1 C3435T Genotype Distributions in Ecuadorians
Authors: Sinués Blanca , Vicente Jorge , Fanlo Ana , Mayayo-Sinués Esteban , González-Andrade Fabricio , Sánchez-Q Dora , Martínez-Jarreta Begoña
Published: June 2013

Polymorphisms in CYP3A genes, such as CYP3A5 and CYP3A4, as well as in the MDR1 gene, which encodes for P‐glycoprotein, have been implicated as genetic markers in several disorders. Differences in the frequency distribution of the allelic variants CYP3A5*3, CYP3A4*1B, and MDR1 3435T have been demonstrated between distinct ethnic groups. In this study we examined the frequency of these allelic variants in 317 healthy Mestizo individuals from Ecuador and made comparisons with results reported in the literature. The genotypes were determined by PCR‐RFLP. Allele and genotype differences were studied by chi‐square test. The MDR1 T allele frequency was similar to that of Spaniard or Asian populations, which is consistent with the ethnic origin of Ecuadorian Mestizo individuals (Amerindian and Spaniard Caucasians). By contrast, the CYP3A5*3 allele frequency was significantly lower in Ecuadorians than in Spaniards and other white populations and higher than in Central Americans, Asians and blacks. CYP3A4*1B was more common in Ecuadorians than in Caucasian or Asian populations but less present than in blacks. The differences in the polymorphism found in this work should be considered in allele‐disease association studies.
Ischemia‐Modified albumin level in type 2 diabetes mellitus—Preliminary report
Authors: Piwowar Agnieszka , Knapik-Kordecka Maria , Warwas Maria
Published: June 2013

Aim: The main goal of the present study was the evaluation of ischemia‐modified albumin (IMA) in patients with type 2 diabetes mellitus and estimation of its connection with vascular complications, glycemic control, hypertension, dyslipidemia and obesity.

Methods: In 76 diabetic patients and 25 control subjects, a plasma level of IMA by manually performed, spectrophotometric Co(II)‐albumin binding assay was determined. Other parameters such as glucose, fructosamine, HbA1c, total cholesterol and its fractions (HDL, LDL), triglicerydes were estimated by routine methods.

Results: Diabetic patients had significantly higher level of IMA in comparison with control subjects. There were not significant differences between groups with various states of vascular complications although the lowest concentration of IMA was observed in patients with microangiopathy. Patients with poor glycemic control had higher IMA level in comparison with these with good glycemic control. Significant correlation was observed between IMA and HbA1c. Among the risk factors, only blood pressure and LDL showed a weak relationship with IMA level.

Conclusions: Our results revealed, for the first time, higher level of IMA in diabetic patients which confirms that it may be of non‐cardiac origin. We can suggest that the albumin molecule in plasma of diabetic patients is modified in the chronic hypoxia conditions provoked mainly by hyperglycemia and oxidative stress in diabetes.
Altered Glycosylation in Tumours Focused to Cancer Diagnosis
Authors: Peracaula Rosa , Barrabés Sílvia , Sarrats Ariadna , Rudd Pauline M. , de Llorens Rafael
Published: June 2013

The lack of specific and sensitive tumour markers for early detection of cancer is driving a search for new approaches that could identify biomarkers. Markers are needed to alert clinicians at the early stages of tumourogenesis, before the cancer has metastasized, when the therapeutic drugs are more effective. Most tumour markers currently used in clinics are serum glycoproteins, frequently highly glycosylated mucins. Typically, the disease marker is the protein and not the glycan moiety of the corresponding glycoprotein or mucin. The increasing knowledge of the role of glycans in cancer suggests that further studies may assist both in determining their role in every step of tumour progression, and in the design of new therapeutic and diagnosic approaches. Detection of the altered glycans in serum tumour glycoproteins could be a way to achieve specificity in tumour detection. In this review, we focus on the glycan changes of two serum glycoproteins, prostate specific antigen ‐ currently used as a tumour marker of prostate cancer ‐ and human pancreatic ribonuclease in pancreatic adenocarcinoma. The detection of glycan changes, associated with subsets of glycoforms in serum glycoproteins that are specific to the tumour situation, could be the basis for developing more specific biomarkers.
Homogeneous Assay of rs4343, an ACE I/D Proxy, and an Analysis in the British Women’s Heart and Health Study (BWHHS)
Authors: Abdollahi Mohammad Reza , Huang Shuwen , Rodriguez Santiago , Guthrie Philip Alexander Isles , Smith George Davey , Ebrahim Shah , Lawlor Debbie A. , Day Ian N. M. , Gaunt Tom R.
Published: June 2013

Current literature suggests that ACE SNP rs4343, ACE 2350A>G in exon 17, T202T, may be the best proxy for the ACE Alu I/D whereas rs4363 and rs4362 may be slightly stronger predictors of ACE levels. Considering reported difficulties in genotyping ACE I/D and stronger associations of rs4343 than ACE I/D with plasma ACE levels in Africans, and suitability of rs4343 for allelic mRNA (cDNA) studies, we developed and validated a liquid phase assay for rs4343, which has advantage on both functional and technical grounds. We confirmed that rs4343, is in near perfect linkage disequilibrium (D’=1, r2 =0.88, n=64) with ACE I/D in Europeans (A and G alleles of rs4343 marking insertion and deletion alleles of ACE I/D respectively).

We then studied its association with metabolic and cardiovascular traits in 3253 British women (60–79 years old).

Apart from a nominal trend of association with diastolic blood pressure (p anova=0.08; p trend=0.05), no other associations were observed. A post‐hoc vascular and general phenome scan revealed no further associations.

We conclude that ACE I/D is not a major determinant of metabolic and cardiovascular traits in this population. Liquid phase genotyping of SNP rs4343 may be preferable to gel based ACE I/D genotyping both for technical and functional reasons.
Molecular Network Analysis of T‐Cell Transcriptome Suggests Aberrant Regulation of Gene Expression by NF‐κB As a Biomarker for Relapse of Multiple Sclerosis
Authors: Satoh Jun-ichi , Misawa Tamako , Tabunoki Hiroko , Yamamura Takashi
Published: June 2013

Molecular mechanisms responsible for acute relapse of multiple sclerosis (MS) remain currently unknown. The aim of this study is to identify the relapse‐specific biomarker genes in T lymphocytes of relapsing‐remitting MS (RRMS). Total RNA of CD3+ T cells isolated from six RRMS patients taken at the peak of acute relapse and at the point of complete remission was processed for DNA microarray analysis. We identified a set of 43 differentially expressed genes (DEG) between acute relapse and complete remission. By using 43 DEG as a discriminator, hierarchical clustering separated the cluster of relapse from that of remission. The molecular network of 43 DEG investigated by KeyMolnet, a bioinformatics tool for analyzing molecular interaction on the curated knowledge database, showed the most significant relationship with aberrant regulation of gene expression by the nuclear factor‐kappa B (NF‐κB) in T cells during MS relapse. These results support the logical hypothesis that NF‐κB plays a central role in triggering molecular events in T cells responsible for induction of acute relapse of MS, and suggest that aberrant gene regulation by NF‐κB on T‐cell transcriptome might serve as a molecular biomarker for monitoring the clinical disease activity of MS.
Cannabinoid Type‐1 Receptor Gene Polymorphisms Are Associated with Central Obesity in a Southern Brazilian Population
Authors: Jaeger Janaína P. , Mattevi Vanessa S. , Callegari-Jacques Sidia M. , Hutz Mara H.
Published: June 2013

The CB1 cannabinoid receptor and its endogenous ligands, the endocannabinoids, are involved in energy balance control, stimulating appetite and increasing body weight in wasting syndromes. Different studies have investigated the relationship between polymorphisms of the cannabinoid receptor 1 (CNR1) gene and obesity with conflicting results. In the present study, we investigated the 1359G/A (rs1049353), 3813A/G (rs12720071) and 4895A/G (rs806368) polymorphisms in the CNR1 gene in a Brazilian population of European descent. To verify the association between these variants and obesity‐related traits in this population, 756 individuals were genotyped by PCR‐RFLP methods. The 4895G allele was associated with waist to hip ratio (WHR) (P = 0.014; P = 0.042 after Bonferroni correction). An additive effect with the GAA haplotype was associated with WHR (P = 0.028), although this statistical significance disappeared after Bonferroni correction (P = 0.084). No significant association was observed between the genotypes of the 1359G/A and 3813A/G polymorphisms and any of the quantitative variables investigated. Our findings suggest that CNR1 gene polymorphism is associated with central obesity in this Brazilian population of European ancestry.
Androgen‐Forming Stem Leydig cells: Identification, Function and Therapeutic Potential
Authors: Zhang Yunhui , Ge Renshan , Hardy Matthew P.
Published: June 2013

Leydig cells are the primary source of testosterone in the male, and differentiation of Leydig cells in the testes is one of the primary events in the development of the male body and fertility. Stem Leydig cells (SLCs) exist in the testis throughout postnatal life, but a lack of cell surface markers previously hindered attempts to obtain purified SLC fractions. Once isolated, the properties of SLCs provide interesting clues for the ontogeny of these cells within the embryo. Moreover, the clinical potential of SLCs might be used to reverse age‐related declines in testosterone levels in aging men, and stimulate reproductive function in hypogonadal males. This review focuses on the source, identification and outlook for therapeutic applications of SLCs. Separate pools of SLCs may give rise to fetal and adult generations of Leydig cell, which may account for their observed functional differences. These differences should in turn be taken into account when assessing the consequences of environmental pollutants such as the phthalate ester, diethylhexylphthalate (DEHP).
Matriptase and Survivin Expression Associated with Tumor Progression and Malignant Potential in Breast Cancer of Chinese Women: Tissue Microarray Analysis of Immunostaining Scores with Clinicopathological Parameters
Authors: Tsai Wen-Chiuan , Chu Chi-Hong , Yu Cheng-Pin , Sheu Lai-Fa , Chen Ann , Chiang Hung , Jin Jong-Shiaw
Published: June 2013

Objective: The aim of this study was to examine the expression of matriptase and survivin in breast carcinoma and correlate with clinicopathological parameters.

Methods: Immunohistochemical analysis of matriptase and survivin were performed in tissue microarray slides of 290 cases, including 11 normal breast tissue; 27 fibrocystic disease; 17 fibroadenoma; 6 atypical ductal hyperplasia; 39 ductal carcinoma in situ, low grade (DCIS, low grade); 39 ductal carcinoma in situ, high grade (DCIS, high grade); 27 invasive ductal carcinoma, grade I (IDC, grade I); 78 invasive ductal carcinoma, grade II (IDC, grade II); and 46 invasive ductal carcinoma, grade III (IDC, grade III).

Results: The average immunostaining scores of matriptase were 44.1 in normal breast tissue, 52.7 in fibrocystic disease, 76.5 in fibroadenoma, 81.7 in atypical ductal hyperplasia, 133.7 in low‐grade DCIS, and 155.8 in high‐grade DCIS. Among 151 breast IDC cases, the average immunostaining scores of matriptase were 172.7 in grade I, 211.7 in grade II, and 221.2 in grade III. Additionally, the average immunostaining scores of surviving also correlate with tumor grades and stages.

Conclusions: Higher expressions of matriptase and survivin correlate significantly with clinicopathological parameters in breast cancer and the malignant potential in premalignant lesions. In addition, higher survivin expression had poorer prognosis of breast IDC cases.
Determination of Glutamic Acid Decarboxylase (GAD65) in Pancreatic Islets and Its In Vitro and In Vivo Degradation Kinetics in Serum Using a Highly Sensitive Enzyme Immunoassay
Authors: Schlosser Michael , Walschus Uwe , Klöting Ingrid , Walther Reinhard
Published: June 2013

Glutamic acid decarboxylase GAD65 autoantibodies (GADA) are an established marker for autoimmune diabetes. Recently, the autoantigen GAD65 itself was proposed as biomarker of beta‐cell loss for prediction of autoimmune diabetes and graft rejection after islet transplantation. Therefore, the GAD65 content in pancreatic islets of different species and its serum degradation kinetics were examined in this study using a sensitive immunoassay. GAD65 was found in quantities of 78 (human), 43.7 (LEW.1A rat) and 37.4 (BB/OK rat) ng per 1,000 islets, respectively, but not in mouse islets. The in vitro half‐life of porcine GAD65 and human recombinant GAD65 ranged from 1.27 to 2.35 hours at 37°C in human serum, plasma and blood, and was unaffected by presence of GAD65 autoantibodies. After injecting 2,000 ng recombinant human GAD65 into LEW.1A rats, the in vivo half‐life was 2.77 hours. GAD65 was undetectable after 24 hours in these animals, and for up to 48 hours following diabetes induction by streptozotocin in LEW.1A rats. Estimated from these data, at least 13 islets in rat and 1,875 in human must be simultaneously destroyed to detect GAD65 in circulation. These results should be taken into consideration in further studies aimed at examining the diagnostic relevance of GAD65.
Racial Differences in Serum Cotinine Levels of Smokers
Authors: Signorello Lisa B. , Cai Qiuyin , Tarone Robert E. , McLaughlin Joseph K. , Blot William J.
Published: June 2013

The purpose of this study was to estimate black/white differences in cotinine levels for current smokers of both sexes, and to explore the potential contribution of mentholated cigarettes to these differences. Sera from 255 current smokers sampled from Southern Community Cohort Study participants (65 black men, 65 black women, 63 white men, 62 white women) were analyzed for cotinine, and linear regression was used to model the effect of race on cotinine level, adjusting for the number of cigarettes smoked within the last 24 hours, use of menthol vs. non-menthol cigarettes, exposure to environmental tobacco smoke, and age. Black smokers smoked fewer cigarettes than white smokers, yet had crude mean cotinine levels nearly as high or higher than white smokers. After multivariate adjustment, cotinine levels were an average of 50 ng/ml higher among black than white women (p=0.008) and non-significantly 12 ng/ml higher among black than white men (p=0.52). We observed no increase in cotinine levels associated with menthol cigarette use. We conclude that differences in cotinine levels among smokers suggest racial variation in exposure to and/or metabolism of tobacco smoke constituents, but our findings do not support a role for menthol preference in this disparity.
Genetic Polymorphisms Modifying Oxidative Stress Are Associated with Disease Activity in Rheumatoid Arthritis Patients
Authors: Bohanec Grabar Petra , Logar Dušan , Tomšič Matija , Rozman Blaž , Dolžan Vita
Published: June 2013

Reactive oxygen and nitrogen species are involved in the pathology of rheumatoid arthritis (RA). Polymorphisms in genes coding for superoxide dismutases (SOD2 and SOD3), catalase (CAT), tumor necrosis factor-alpha (TNFA) and inducible NO synthase (NOS2A) may influence RA activity. We determined SOD2 Ala-9Val, SOD3 Arg213Gly, CAT C-262T, TNFA G-308A, TNFA C-857T and NOS2A (CCTTT)npolymorphisms in 327 RA patients. Carriers of CAT -262T and TNFA -308A allele had lower mean disease activity score of 28 joint count (DAS28) values than patients with CAT -262CC and TNFA -308GG genotypes (p= 0.014 andp= 0.046, respectively). Patients with the combination of CAT -262T and TNFA -308A allele had lower mean DAS28 values and a higher probability for low disease activity than non-carriers (p= 0.003, OR = 3.585, 95% CI = 1.538–8.357). Our results suggest that CAT and TNFA polymorphisms alone and in combination influence the activity of RA.
Variability of Thiobarbituric Acid Reacting Substances in Saliva
Authors: Behuliak Michal , Pálffy Roland , Gardlík Roman , Hodosy Július , Halčák Lukáč , Celec Peter
Published: June 2013

Introduction: Salivary TBARS are a potential marker of oxidative stress in the oral cavity. Previous studies have found increased levels of salivary TBARS in various diseases. The aim of this study was to assess the variability of salivary TBARS in both genders.

Subjects & Methods: Saliva samples from thirty-eight healthy volunteers (18F & 20M) were collected every day during 30 day period. TBARS levels were measured spectrophotometrically using a high-throughput 96-well plate method. Time series analysis was performed using standard statistical methods.

Results: Repeated measures ANOVA showed a significant variation of salivary TBARS within day and subjects (p< 0.001). The dynamics did not differ between genders. Intraindividual variability was very high in both genders with coefficients of variation of more than 60%. Interindividual variability was higher in men than in women (73% vs. 46%;p< 0.01).

Discussion: The relatively high intraindividual variability indicates that the use of salivary TBARS will be limited to research on a population level, although some informative value might be gained by repeated samplings. Factors influencing the biological variability of salivary TBARS should be identified in further studies.
Peptide Bound Hypohydroxyprolinuria in Handigodu Disease: A Familial Syndrome of Spondylo Epi(meta)physeal Dysplasia
Authors: Badadani Mallikarjun , Babu S. V. Suresh , Shetty K. T. , Agarwal S. S.
Published: June 2013

Handigodu Disease (HD) is disorder of the osteoarticular system prevalent in few villages of two districts of the state Karnataka in southern India. 24 hrs urinary excretions of proline (Pro) and 4-hydroxyproline (Hyp) were analyzed by HPLC. Decreased peptide bound Hyp excretions (μmole/24 hrs) were found in patient group when compared with controls (Nonaffected; 113.02 ± 67.96, Type-I; 36.22 ± 20.76, Type-II; 45.74 ± 14.95, Type-III; 40.46 ± 22.68) and without significant difference in Pro excretions. Significant increased peptide bound Pro to Hyp ratio were found in patient group compared to control (Nonaffectedn= 63: 2.02 ± 1.65, Type-In= 18: 3.144 ± 1.42, Type-IIn= 28: 4.21 ± 1.95, Type-IIIn= 8: 8.60 ± 6.55). 24 hrs urinary excretions of deoxypyridinoline (DPD) crosslinks were found without significant difference among affected and control, hence HD ruled out from general bone reduction. These results suggest hypohydroxyprolinuria may be because of reduced bone turnover or defective hydroxylation of prolyl residues during post translational modification of collagen biosynthesis.
Frequency of the Hemochromatosis Gene (HFE) Variants in a Jordanian Arab Population and in Diabetics from the Same Region
Authors: Alkhateeb Asem , Uzrail Amal , Bodoor Khaldon
Published: June 2013

Hereditary HFE-linked hemochromatosis is a frequent recessive disorder among individuals of northern European ancestry. The clinical characteristic of this disease is the gradual accumulation of iron in internal organs, which ultimately may lead to organ damage and death. Three allelic variants of HFE gene have been correlated with hereditary hemochromatosis: C282Y is significantly associated with hereditary hemochromatosis in populations of Celtic origin, H63D and S65C are associated with milder form of iron overload. In this study we performed mutation analysis to identify allele frequency of the three variants of HFE gene in Jordanian Arab population, to assess deviations of these frequencies from those detected elsewhere, and to determine if there is an increased frequency of these variants in a diabetic population (Type 2 diabetes) from the same area. DNA was extracted from blood samples of 440 individuals attending King Abdullah University Hospital for ambulatory services. We used polymerase chain reaction (PCR) to amplify exons 2 and 4 of the HFE gene then restriction fragment length polymorphism (RFLP) method to detect the variants. There were neither homozygous nor heterozygous for C282Y variant. For the H63D variant, 0.68% were homozygous and 21.1% were heterozygous. For the S65C variant, there were no homozygous and 0.23% were heterozygous. Allelic frequencies were, 0%, 11.25%, and 0.11% for C282Y, H63D, and S65C, respectively. Our samples were subdivided into two categories of type 2 diabetic (89 cases) and controls (blood donors, 204 cases) and compared with regard to the H63D variant. Both groups did not have homozygous H63D variant. H63D heterozygous in diabetics were 23.60% and in blood donor controls 22.55%. Allelic frequency of the mutant H63D allele was 11.80% in diabetics and 11.27% for the blood donor controls. This is the first study to show the frequency of the three hemochromatosis gene variants in Jordan with the interesting finding of no C282Y allele detected in 440 samples. Additionally, no significant difference was observed in H63D variant frequency in type 2 diabetics as compared to controls.
A Systematic Review of Studies Examining Inflammation Associated Cytokines in Human Abdominal Aortic Aneurysm Samples
Authors: Golledge Alexandra L. V. , Walker Philip , Norman Paul E. , Golledge Jonathan
Published: June 2013

Objectives: Inflammation is critical in abdominal aortic aneurysm (AAA) but there is no current consensus on which inflammation related cytokines are important. The aim of this review was to systemically assess previous studies investigating the relative expression of inflammation associated cytokines within human AAA samples.

Methods: The MEDLINE database was searched for studies which simultaneously examined an array of different inflammation associated cytokines in aortic samples in order to identify those associated with AAA. Focused searches were then conducted for further studies assessing relative concentrations of these cytokines in aortic samples in relation to AAA. Appropriate studies were assessed by two reviewers independently.

Results: Eighteen studies were included. A number of different cytokines have been consistently found to be upregulated within AAA by comparison to aortic samples removed from patients without cardiovascular disease, however findings relative to samples of aortic athero-thrombosis were less consistent. TNFA and INFG appear to be the most consistently associated with AAA in studies using both normal and atherosclerotic controls. Cautious interpretation of these data is recommended due to a number of methodological problems.

Conclusions: This systematic review suggests that TNFA and INFG are the most consistently upregulated cytokines in large AAAs. Further studies utilizing larger populations, new proteomic techniques and better patient matching are required.
A Splice Variant ofGNB3and Peripheral Polyneuropathy in Type 1 Diabetes
Authors: Chistiakov Dimitry A. , Spitsina Ekaterina V. , Nikitin Alexey G. , Strokov Igor A. , Nosikov Valery V.
Published: June 2013

Abnormalities in G protein-mediated signal transduction could be involved in the pathogenesis of diabetic polyneuropathy (DPN). Here we test whether the GNB3 C825T variant confers susceptibility to DPN in type 1 diabetes (T1D) mellitus. The C825T marker of GNB3 was genotyped in genomic DNA from blood isolated from a total of 213 Russian T1D patients 100 of whom had DPN. Compared to carriers of the wild-type genotypeC/C, diabetic subjects with genotypesT/Thad significantly increased risk to develop DPN (Odds Ratio (OR) of 4.4 (p= 0.001). The adjustment for confounders (age, sex, body mass index, cigarette smoking, and level of reduced glutathione) resulted in increase of the OR value up to 4.72 (p= 8.9 × 10-3). The further adjustment for hypertension abolished the association between the GNB3 C825T variant and DPN (OR = 1.95,p= 0.18). Non-complicated subjects homozygous forT/Tshowed decreased levels of reduced glutathione (T/T: 69 ± 19 vs.C/T: 74 ± 19 vs.C/C: 77 ± 17 μmol/l,p= 0.009). Compared to other GNB3 variants, carriers of theT/Tgenotype had elevated systolic blood pressure (SBP) in complicated (T/T: 115.8 ± 9.1 vs.C/T: 113.3 ± 8.2 vs.C/C: 109.5 ± 8.7 mm/Hg,p= 0.036) and non-complicated T1D patients (T/T: 118.1 ± 8.4 vs.C/T: 116.9 ± 7.9 vs.C/C: 112.1 ± 7.2 mm/Hg,p= 0.02). However, the significance of association between the C825T polymorphism was lost after adjustment for confounding risk factors. In conclusion, the 825T allele of GNB3 is likely to accelerate the development of DPN through primary effects to SBP and hypertension in subgroups of diabetic patients with impaired neurovascular function and advanced oxidative stress.
Genetic Factors Associated with Rheumatoid Arthritis and Systemic Vasculitis: Evaluation of a Panel of Polymorphisms
Authors: Menegatti Elisa , Davit Annalisa , Francica Simona , Berardi Daniela , Rossi Daniela , Baldovino Simone , Tovo Pier Angelo , Sena Luigi M. , Roccatello Dario
Published: June 2013

Immune and inflammatory response activation is a common feature of connective tissue diseases and systemic vasculitis. The aim of our study was to evaluate the possible involvement of TNFα c.-308A > G, IL-10 c.-1082A > G, uteroglobin c.38A > G, TGFβ 1 c.869C > T and NFκB2 c.-1837T > C gene polymorphisms in susceptibility to connective tissue diseases. Our study cohort included 68 unrelated patients affected by rheumatoid arthritis (RA) (37 patients) and ANCA-positive [micropolyangiitis (mPA) 17 patients] or ANCA-negative systemic vasculitis [including 8 patients with Henoch-Schönlein purpura (HSP) and 6 patients with mixed cryoglobulinaemia (MC)] as well as 98 control subjects. Allele frequency analysis of uteroglobin c.38G > A polymorphism showed a significant increase in the c.38A allele in patients (p= 0.002). Genotype frequency analysis of uteroglobin and NF-κB2 gene polymorphisms in patients showed an increase in c.38GA and c.38AA genotypes in the uteroglobin gene (p=0.02) coupled with an increase in homozygous c.-1837CC in the NF-κB2 gene (p=0.02). Our data suggest that genetic variation in UG and NF-κB2 pathways could have effects in connective tissue disease susceptibility.
Clinical Significance of Inflammatory Markers in Polycystic Ovary Syndrome: Their Relationship to Insulin Resistance and Body Mass Index
Authors: Samy Nervana , Hashim Maha , Sayed Magda , Said Mohamed
Published: June 2013

Background: Women with polycystic ovary syndrome (PCOS) have an increased prevalence of insulin resistance (IR) and related disorders. Elevated serum levels of high sensitivity CRP (hs-CRP), interleukin-6 (IL-6) and tumor necrosis factorα(TNF-α) reflect low-grade chronic inflammation and have been associated with several insulin-resistant states; they are useful cardiovascular risk markers. The objective of this study was to investigate whether soluble inflammatory markers are altered in PCOS focusing on its relationship with obesity and indexes of insulin resistance.

Patients and methods: One hundred and eight women with PCOS and 75 healthy women were recruited. Patients were divided according to body mass index (BMI) into two groups; group I (BMI < 27 kg/m2) and group II (BMI ≥ 27 Kg/m2). Serum levels of hs-CRP, IL-6, and TNF-α, lipid and hormone profiles were measured.

Results: PCOS patients had increased levels of testosterone, luteinizing hormone (LH), androstendione, insulin level and HOMA index compared to healthy BMI matched controls. High-density lipoprotein (HDL) concentrations were significantly reduced in both patient groups compared to their controls, while triglyceride levels were significantly increased in obese group compared to controls. There were no significant difference in serum inflammatory markers hs-CRP, IL-6 and TNF-αbetween group I and their matched controls. On the other hand, there were significant increase in these markers between group II and their matched controls. There were highly significant positive correlation between hs-CRP and IL-6 (r= 0.702,P< 0.001) and between hs-CRP and TNF-α(r= 0.621,P<0.001), also between IL-6 and TNF-α(r= 0.543,P< 0.001). These inflammatory markers correlated significantly with BMI and HOMA index. Multiple regression analysis revealed that BMI and HOMA were predictors of IL-6 levels (b= 11.173,P< 0.001,b= 13.564,P< 0.001 respectively) and BMI was the only predictor of hs-CRP levels (b= 12.578,P< 0.001) and TNF-αlevels (b= 0.134,P< 0.001).

Conclusion: PCOS and obesity induce an increase in serum inflammatory cardiovascular risk markers. The precise mechanisms underlying these associations require additional studies to clarify the state of the cardiovascular system in women with PCOS compared with controls in large numbers of patients to determine the relative contribution of different factors including insulin resistance, androgen status and BMI.
suPAR: The Molecular Crystal Ball
Authors: Thunø Maria , Macho Betina , Eugen-Olsen Jesper
Published: June 2013

soluble urokinase Plasminogen Activator Receptor (suPAR) levels reflect inflammation and elevated suPAR levels are found in several infectious diseases and cancer. suPAR exists in three forms; suPARI-III, suPARII-IIIand suPARIwhich show different properties due to structural differences. Studies suggest that full-length suPAR is a regulator of uPAR/uPA by acting as uPA-scavenger, whereas the cleaved suPARII-IIIact as a chemotactic agent promoting the immune response via the SRSRY sequence in the linker-region. This review focus on the various suPAR fragments and their involvement in inflammation and pathogenic processes. We focus on the molecular mechanisms of the suPAR fragments and the link to the inflammatory process, as this could lead to medical applications in infectious and pathological conditions.
Variations in Serum Transforming Growth Factor-β1 Levels with Gender, Age and Lifestyle Factors of Healthy Japanese Adults
Authors: Lin Yingsong , Nakachi Kei , Ito Yoshinori , Kikuchi Shogo , Tamakoshi Akiko , Yagyu Kiyoko , Watanabe Yoshiyuki , Inaba Yutaka , Tajima Kazuo , JACC Study Group
Published: June 2013

Elevated serum or plasma Transforming Growth Factor-β1 (TGF-β1) levels have been linked to cancer and other diseases in numerous studies; however, very few studies have reported an association between circulating TGF-β1 and lifestyle factors in healthy people. We examined the association between serum TGF-β1 levels and gender, age, body mass index (BMI), smoking, and drinking in a large population-based cohort study (N = 9,142). Serum TGF-β1 levels were detected by the Quantikine enzyme-linked immunoassay kit (R&D Systems). The data indicated highly significant (p<0.0001) difference in serum TGF-β1 levels between men (mean value: 37.6 ± 0.12 ng/mL, N = 4888) and women (mean value: 35.1 ± 0.12 ng/ml, N = 4254). Serum TGF-β1 levels decreased with age (trend p < 0.0001) and were positively associated with obesity (trend p < 0.0001) in both men and women. We observed a significant trend with increased serum TGF-β1 levels corresponding to increased amount of tobacco and alcohol consumption in men (trend p < 0.0001). These findings suggest that serum TGF-β1 levels appear to be modulated by gender, age and lifestyle factors such as obesity, cigarette smoking, and alcohol drinking in healthy Japanese adults.
Estrogen Receptor-αgene (T/C) Pvu II Polymorphism in Endometriosis and Uterine Fibroids
Authors: Govindan Sujatha , Shaik Noor Ahmad , Vedicherla Bhavani , Kodati Vijayalakshmi , Rao Kaipa Prabhakar , Hasan Qurratulain
Published: June 2013

Endometriosis and fibroids are estrogen-dependent benign pathologies of the uterus, which account for infertility and pelvic pain along with dysmenorrhea in women. Suppression of the disease and recurrence after discontinuing hormone therapy strongly suggests that these are responsive to hormones, especially estrogen, which acts via its receptor. A T/C SNP in intron 1 and exon 2 boundary of estrogen receptor (ER)αgene recognized by PvuII enzyme has been associated with several female pathologies like breast cancer, osteoporosis, endometriosis and fibroids in various ethnic groups. The aim of the present study was to assess this ERαT/C polymorphism in endometriosis and fibroid patients from Asian Indian population. Genomic DNA was isolated from 367 women, who included 110 cases of endometriosis, 142 cases of uterine fibroids and 115 healthy age matched women volunteers. PCR was carried out to amplify ERαgene followed by restriction digestion with Pvu II. Results indicate a significant association of C allele with both endometriosis [OR = 2.6667, 95% CI = 1.4166 to 5.0199;p< 0.05] and fibroids [2.0833, 95% CI = 1.1327 to 3.8319;p< 0.05]. Further studies are needed in larger population to establish ERαC allele as a risk marker for endometriosis and fibroids in Asian Indian women. Ethnicity, race, diet etc may play a role in susceptibility to endometriosis and fibroids and further studies are warranted in this area.
TPMT and DPD Polymorphisms: Efficient Screening Method for Indian Patients Considering Taking Thiopurine and 5-FU Drugs
Authors: Ashavaid T. F. , Raghavan Rani , Shah Swarup , Kapadia Asha , Almel Sachin , Desai Devendra
Published: June 2013

Introduction: Development of DNA-based tests for TPMT/DPD polymorphisms can help clinicians and patients to make important decisions about cancer treatment. Also, due to lack of Indian data, we aimed at the development and validation of these tests in Indian patients.

Materials and Methods: Molecular assays were used for identifying TPMT/DPD variations; validated by DNA sequencing.

Results: Molecular assays have been used for screening TPMT∗2,∗3A,∗3B,∗3C alleles and IVS14+1(G→A) in DPD gene. A patient, exhibiting neutropenia on 6-MP was observed to be G460A-homozygote, while, two Acute Lymphoblastic Leukemia (ALL) patients with side-effects exhibited wild-type alleles. Two patients showing 6-MP side-effects and responding well to the same drug at later stage also carried wild-type alleles.

Discussion: G460A homozygosity in a patient allowed clinicians to stop 6-MP treatment, improving patient's health status. Two ALL patients showing side-effects were wild-type, indicating presence of unidentified rare variations. Two patients with wild-type allele showed side-effects during 6-MP treatment, but responded well to same drug at later stage, suggesting side-effects to be attributable to multiple biological and environmental processes. Absence of IVS14+1(G→A) in DPD gene will not exclude possibility of another mutation.

Conclusion: Molecular assays for determining common TPMT/DPD variations, can provide accurate diagnosis and efficient therapies in future clinical studies.
MicroRNA Microarray Expression Profiling in Human Myocardial Infarction
Authors: Boštjančič Emanuela , Zidar Nina , Glavač Damjan
Published: June 2013

MicroRNAs (miRNAs), small non-coding RNA molecules, are negative regulators of gene expression. Recent studies have indicated their role in various forms of cardiovascular disease. In spite of the number of miRNA microarray analyses performed, little is known about the genome-wide miRNA expression pattern in human myocardial infarction (MI). Using miRNA microarrays and bioinformatic analysis, miRNA expression was analyzed on human MI and foetal hearts compared to healthy adult hearts, to determine whether there is any similar expression pattern between MI and foetal hearts, and to identified miRNAs that have not previously been described as dysregulated in cardiovascular diseases. Of 719 miRNAs analyzed, ∼ 50% were expressed in human hearts, 77 miRNAs were absent from all tested tissues and 57 were confidently dysregulated in at least one tested group. Some expression patterns appeared to be similar in MI and foetal hearts. Bioinformatic analysis revealed 10 miRNAs as dysregulated in MI not yet related to cardiovascular disease, and 5 miRNAs previously described only in animal models of cardiovascular diseases. Finally, qRT-PCR analysis confirmed dysregulation of 7 miRNAs,miR-150, miR-186, miR-210, miR-451, and muscle-specific, miR-1 and miR-133a/b; all of these are believed to be involved in various physiological and pathological processes.
Emerging Role of Endothelial and Inflammatory Markers in Preeclampsia
Authors: Swellam Menha , Samy Nervana , Abdl Wahab Susan , Ibrahim Mohamed Saeed
Published: June 2013

Objectives:Endothelial disturbance and excess inflammatory response are pathogenic mechanisms in pre-eclampsia (PE). Authors determine the clinical diagnostic role for thrombomodulin (TM), plasminogen activator inhibitor-1 (PAI-1) as endothelial markers and C-reactive protein (CRP), and interlukin-6 (IL-6) as inflammatory markers when tested independently or in combinations.

Materials and methods:We conducted a retrospective study in a cohort of 185 women grouped as 80 women with PE, 55 normotensive pregnant and 50 healthy non-pregnant. Plasma levels of TM, PAI-1, CRP and IL-6 were examined using enzyme linked immunosorbent assays.

Results:Median levels and the positivity rates for the investigated markers were higher in PE as compared to the other groups (P< 0.0001). Using linear regression analysis, the investigated markers were significantly correlated regarding healthy nonpregnantvsPE or normotensive pregnantvsPE. The sensitivity of PAI-1 was the highest (98%) among the tested biomarkers. Combination between the investigated markers revealed absolute sensitivity (100%) and reliable specificity especially when PAI-1 was combined with CRP at 83% specificity.

Conclusions:Investigated endothelial and inflammatory markers revealed sensitive diagnostic test for PE. However, coupled combination between PAI-1 with CRP showed superior both sensitivity and specificity which represent a promising new approach for detection of PE.