bluuebunny/crossref_metadata_2025 huggingface dataset:
Human GST Loci as Markers of Evolutionary Forces: GSTO1*E155del and GSTO1*E208K Polymorphisms May Be Under Natural Selection Induced by Environmental Arsenic
Authors: Polimanti Renato , Piacentini Sara , De Angelis Flavio , De Stefano Gian Franco , Fuciarelli Maria
Published: June 2013

Over the last two decades, significant data has been accumulated linking Glutatione S-Transferases (GSTs) with the development of several diseases. Contemporary studies have demonstrated the impact of ethnicity on GST allele frequencies. The aim is to verify if the variability of GST genes reflects population demographic history or rather selective pressures.

GST genes (GSTM1, GSTO1 GSTO2, GSTT1) were analysed in three Ecuadorian populations (Cayapas,n= 114; Colorados,n= 104; African-Ecuadorian,n= 77) and compared with HapMap data. GST SNPs were determined using the PCR-RFLP method while GST null phenotype was determined using a Multiplex PCR.

The population relationship achieved using GSTM1 positive/null, GSTO1*A140D, GSTO2*N142D and GSTT1 positive/null are in agreement with the data obtained using neutral polymorphisms: Amerindians are close to Asian populations and African-Ecuadorians to African populations. To what concerns GSTO1*del155 and GSTO1*K208 variants, allele frequencies never exceeded 10%, showing no significant differences in the Ecuadorian groups and in worldwide populations.

The features of GSTO1*del155 and GSTO1*K208 variants and their association with arsenic biotransformation deficiency suggest the presence of a selection mechanism towards these loci. In particular, this hypothesis is strengthened by a possible linkage between these alleles and the susceptibility of arsenic-induced male infertility.
Serum Progranulin as an Independent Marker of Liver Fibrosis in Patients with Biopsy-Proven Nonalcoholic Fatty Liver Disease
Authors: Yilmaz Yusuf , Eren Fatih , Yonal Oya , Polat Zulfikar , Bacha Mohammad , Kurt Ramazan , Ozturk Oguzhan , Avsar Erol
Published: June 2013

Background:Elevated progranulin levels are associated with visceral obesity, elevated plasma glucose, and dyslipidemia. Progranulin has not been previously investigated as a biomarker of nonalcoholic fatty liver disease (NAFLD). We sought to determine whether serum progranulin levels are altered in patients with biopsy-proven NAFLD and if they are associated with their clinical, biochemical, and histological characteristics.

Subjects and methods:We measured serum progranulin levels in 95 patients with biopsy-proven NAFLD and 80 age- and sex-matched controls. The potential associations between progranulin and the characteristics of NAFLD patients were examined by multiple linear regression analysis.

Results:Serum progranulin levels were significantly higher in NAFLD patients (34 ± 13 ng/mL) than in controls (28 ± 7 ng/mL,P< 0.001). In NAFLD patients, serum progranulin levels were associated with lipid levels and the degree of hepatic fibrosis. After adjustment for potential confounders, serum progranulin remained an independent predictor of the degree of hepatic fibrosis in NAFLD patients (β= 0.392;t=2.226,P< 0.01).

Conclusions:Compared with controls, NAFLD patients have higher serum progranulin concentrations, which are closely associated with lipid values and the extent of hepatic fibrosis.
Analysis of Sputum Markers in the Evaluation of Lung Inflammation and Functional Impairment in Symptomatic Smokers and COPD Patients
Authors: Paone Gregorino , Conti Vittoria , Vestri Annarita , Leone Alvaro , Puglisi Giovanni , Benassi Fulvio , Brunetti Giuseppe , Schmid Giovanni , Cammarella Ilio , Terzano Claudio
Published: June 2013

The pivotal role of neutrophils and macrophages in smoking-related lung inflammation and COPD development is well-established.

We aimed to assess whether sputum concentrations of Human Neutrophil Peptides (HNP), Neutrophil Elastase (NE), Interleukin-8 (IL-8), and Metalloproteinase-9 (MMP-9), major products of neutrophils and macrophages, could be used to trace airway inflammation and progression towards pulmonary functional impairment characteristic of COPD.

Forty-two symptomatic smokers and 42 COPD patients underwent pulmonary function tests; sputum samples were collected at enrolment, and 6 months after smoking cessation.

HNP, NE, IL-8, MMP-9 levels were increased in individuals with COPD (p< 0.0001). HNP and NE concentrations were higher in patients with severe airways obstruction, as compared to patients with mild-to-moderate COPD (p= 0.002). A negative correlation was observed between FEV1and HNP, NE and IL-8 levels (p< 0.01), between FVC1/FVC and HNP, NE and IL-8 levels (p< 0.01), and between NE enrolment levels and FEV1decline after 2 years (p= 0.04).

ROC analysis, to discriminate symptomatic smokers and COPD patients, showed the following AUCs: for HNP 0.92; for NE 0.81; for IL-8 0.89; for MMP-9 0.81; for HNP, IL-8 and MMP-9 considered together 0.981.

The data suggest that the measurement of sputum markers may have an important role in clinical practice for monitoring COPD.
Immunological Functions of Oxidized Human Immunoglobulin G in Type 1 Diabetes Mellitus: Its Potential Role in Diabetic Smokers as a Biomarker of Elevated Oxidative Stress
Authors: Rasheed Zafar , Al-Shobaili Hani A. , Alzolibani Abdullateef A. , Ismail Khan Muhammad , Tariq Ayub Muhammad , Khan Mohammed Imran , Rasheed Naila
Published: June 2013

The role of oxidized immunoglobulin G in type 1 diabetic smokers has been investigated in the present study. Human immunoglobulin G (IgG) was modified by reactive oxygen species (ROS). The binding characteristics of circulating autoantibodies in type 1 diabetes patients against native and modified IgG were assessed by direct binding ELISA. High degree of specific binding by 68.5% of patients sera towards ROS-modified IgG was observed in comparison to its native analogue (p< 0.05). In addition, diabetic smokers (n= 28) were examined and the results were compared with diabetic non-smokers (n= 26). Circulating antibodies of diabetic smokers showed substantially stronger binding to modified IgG as compared with the antibodies present in diabetic non-smokers (p< 0.05). Normal human sera (n= 53) showed negligible binding with either antigen. Competitive inhibition ELISA reiterates the direct binding results. The increase in total serum protein carbonyl levels in the diabetic smokers was largely due to an increase in oxidized IgG. Diabetic smokers showed substantially higher carbonyl contents in sera as well as in purified IgG as compared with sera and IgG of diabetic non-smokers. Collectively, the oxidation of plasma proteins, especially IgG, might enhance oxidative stress in type 1 diabetes smokers.
Deregulated Expression of Superoxide Dismutase-2 Correlates with Different Stages of Cervical Neoplasia
Authors: Termini Lara , Filho Adhemar Longatto , Maciag Paulo Cesar , Etlinger Daniela , Alves Venâncio Avancini Ferreira , Nonogaki Suely , Soares Fernando Augusto , Villa Luisa Lina
Published: June 2013

Objective: Superoxide dismutase-2 (SOD2) is considered one of the most important antioxidant enzymes that regulate cellular redox state in normal and tumorigenic cells. Overexpression of this enzyme may be involved in carcinogenesis, particularly in lung, gastric, colorectal and breast cancer.

Methods: In the present study, we have evaluated SOD2 protein levels by immunohistochemistry (IHC) in 331 cervical histological samples including 31 low-grade cervical intraepithelial neoplasia (LSIL), 51 high-grade cervical intraepithelial neoplasia (HSIL), 197 squamous cervical carcinomas (SCC) and 52 cervical adenocarcinomas (ADENO).

Results: We observed that SOD2 staining increases with cervical disease severity. Intense SOD2 staining was found in 13% of LSIL, 25.5% of HSIL and 40% of SCC. Moreover, 65.4% of ADENO exhibited intense SOD2 staining.

Conclusions: Differences in the expression of SOD2 could potentially be used as a biomarker for the characterization of different stages of cervical disease.
Association of Interleukin 23 Receptor Gene with Sarcoidosis
Authors: Kim Hyun Soo , Choi Dongseok , Lim Lyndell L. , Allada Gopal , Smith Justine R. , Austin Carrie R. , Doyle Trudy M. , Goodwin Kelley A. , Rosenbaum James T. , Martin Tammy M.
Published: June 2013

Interleukin 23 receptor (IL23R) gene has been reported as a genetic factor strongly associated with inflammatory bowel disease, psoriasis, and ankylosing spondylitis. We investigated the association betweenIL23Rgene single nucleotide polymorphisms (SNPs) and susceptibility to sarcoidosis, including the clinical manifestation of uveitis.

Ninety-one sarcoidosis subjects (58 with and 33 without uveitis) and 104 healthy controls were genotyped for eleven IL23R SNPs. DNA was amplified using specific PCR primers and genotyped by denaturing HPLC and/or direct DNA sequencing. Case-control frequency comparisons were analyzed using Chi square test.

ThreeIL23RSNPs, rs7517847 (intron 6), rs11465804 (intron 8), and rs11209026 (exon 9, c.1142G>A, p.Arg381Gln) were associated with sarcoidosis in our population (p< 0.05): rs7517847 showed increased frequencies in sarcoidosis compared to controls, but rs11465804 and rs11209026 were decreased. Two of these SNPs were associated with the uveitis subgroup compared to controls: rs11465804 (0.9% vs. 7.2%, OR = 0.11, P = 0.013) and rs11209026 (1.8% vs. 7.3%, OR = 0.23, P = 0.038).

This finding indicates the association ofIL23Rpolymorphism with sarcoidosis, especially with sarcoid uveitis.IL23Rmay be a common susceptibility gene shared by several autoimmune disorders including inflammatory bowel disease, psoriasis, and ankylosing spondylitis and sarcoid uveitis.
The Role of Genes in Defining a Molecular Biology of PTSD
Authors: Yehuda Rachel , Koenen Karestan C. , Galea Sandro , Flory Janine D.
Published: June 2013

Because environmental exposure to trauma is the sine qua non for the development of Post Traumatic Stress Disorder (PTSD), the recent focus on genetic studies has been noteworthy. The main catalyst for such studies is the observation from epidemiological studies that not all trauma survivors develop this disorder. Furthermore, neuroendocrine findings suggest pre-existing hormonal alterations that confer risk for PTSD. This paper presents the rationale for examining genetic factors in PTSD and trauma exposure, but suggests that studies of genotype may only present a limited picture of the molecular biology of this disorder. We describe the type of information that can be obtained from candidate gene and genomic studies that incorporate environmental factors in the design (i.e., gene – environment interaction and gene-environment correlation studies) and studies that capitalize on the idea that environment modifies gene expression, via epigenetic or other molecular mechanisms. The examination of epigenetic mechanisms in tandem with gene expression will help refine models that explain how PTSD risk, pathophysiology, and recovery is mediated by the environment. Since inherited genetic variation may also influence the extent of epigenetic or gene expression changes resulting from the environment, such studies should optimally be followed up by studies of genotype.
High-Resolution Melting Curve Analysis for High-Throughput Genotyping of NOD2/CARD15 Mutations and Distribution of These Mutations in Slovenian Inflammatory Bowel Diseases Patients
Authors: Mitrovič Mitja , Potočnik Uroš
Published: June 2013

Inflammatory bowel diseases (IBD) are usually classified into Crohn's disease (CD) and ulcerative colitis (UC). NOD2/CARD15 was the first identified CD-susceptibility gene and was confirmed as the most potent disease gene in CD pathogenesis. ThreeNOD2/CARD15variants, namely two missense polymorphisms R702W (rs2066844) and G908R (rs2066845), and a frame shift polymorphism L1007fs (rs2066847), were associated with CD in Caucasian populations. High resolution melting analysis (HRMA) with saturation LCGreen dyes was previously reported as a simple, inexpensive, accurate and sensitive method for genotyping and/or scanning of rare variants. For this reasons we used qPCR-HRMA for genotypingNOD2/CARD15variants in 588 Slovenian IBD patients and 256 healthy controls. PCR-RFLP was used as a reference method for genotyping of clinical samples. The optimization of an HRM experiment required careful design and adjustment of main parameters, such as primer concentration, MgCl2concentration, probe design and template DNA concentration. Different HRMA approaches were tested and used to develop a reliable and low-cost SNP genotyping assays for polymorphisms inNOD2/CARD15gene. Direct HRMA was the fastest and cheapest HRMA approach for L1007fs and R702W polymorphisms, yet for G908R polymorphism sufficient reliability was achieved after introduction of unlabeled probe. In association analysis, we found statistically significant association of L1007fs (p = 0.001, OR = 3.011, CI95%=1.494–6.071) and G908R (p = 2.62 × 10-4, OR = 14.117, CI95% = 1.884–105.799) polymorphisms with CD patients. At least one ofNOD2/CARD15polymorphisms was found in 78/354 (22.03%) in CD patients, 25/197 (12.69%) in UC patients and in 26/256 (10.15%) in healthy controls. We have successfully implementedNOD2/CARD15HRMA assays, which may contribute to the development of genetic profiles for risk prediction of developing CD and for differential diagnosis of CD vs. UC.
Immunosenescent CD57+CD4+T-cells Accumulate and Contribute to Interferon-γResponses in HIV Patients Responding Stably to ART
Authors: Fernandez Sonia , French Martyn A. , Price Patricia
Published: June 2013

HIV-infected individuals responding to antiretroviral therapy (ART) after severe CD4+T-cell depletion may retain low responses to recall antigens [eg: cytomegalovirus (CMV)] and altered expression of T-cell co-stimulatory molecules consistent with immunosenescence. We investigated the capacity of phenotypically senescent cells to generate cytokines in HIV patients receiving long-term ART (n= 18) and in healthy controls (n= 10). Memory T-cells were assessed by interferon (IFN)-γELISpot assay and flow cytometrically via IFN-γor IL-2. Proportions of CD57brightCD28nullCD4+T-cells correlated with IFN-γresponses to CMV (p= 0.009) and anti-CD3 (p= 0.002) in HIV patients only. Proportions of CD57brightCD28nullCD8+T-cells and CD8+T-cell IFN-γresponses to CMV peptides correlated in controls but not HIV patients. IL-2 was predominantly produced by CD28+T-cells from all donors, whereas IFN-γwas mostly produced by CD57+T-cells. The findings provide evidence of an accumulation of immunosenescent T-cells able to make IFN-γ. This may influence the pathogenesis of secondary viral infections in HIV patients receiving ART.
HLA and Bronchopulmonary Dysplasia Susceptibility: A Pilot Study
Authors: Rocha Gustavo , Proença Elisa , Areias Augusta , Freitas Fátima , Lima Bruno , Rodrigues Teresa , Alves Helena , Guimarães Hercília
Published: June 2013

There is little data on the association between Human Leucocyte Antigen (HLA) alleles and Bronchopulmonary Dysplasia (BPD) of the preterm newborn. Our aim was to assess associations between HLA alleles and BPD susceptibility. We studied 156 preterm neonates (82 M/74 F) < 32 weeks gestational age, alive at 36 weeks gestational age. Detailed clinical data were collected. HLA typing was performed by PCR-SSO. HLA allele frequencies where determined by direct counting for BPD and no-BPD groups. Comparison between BPD and no BPD groups was performed usingt-test, χ2test or Fisher exact test and logistic regression as appropriate. Relative risks (RR) and their 95% confidence intervals (95% CI) were also calculated as association measures. We diagnosed 56 (35.9%) neonates with mild BPD and 27 (17%) with moderate/severe BPD. We found a significant association between HLA-DRB1*01 and mild BPD (OR=3.48[1.23–10.2]). The alleles HLA-A*24, -A*68, -B*51,-Cw*07, -Cw*14, -Cw*15 and -DRB1*01 presented a significant association with moderate/severe BPD. When adjusted to gestational age and birth weight HLA-A*68 (OR=5.41[1.46; 20.05]), -B*51 (OR=3.09[1.11; 8.63]) and -Cw*14 (OR=4.94[1.15; 21.25]) were significantly associated with moderate/severe BPD. Conclusion – Our findings suggest an association between HLA-A*68, -B*51 and -C*14 and BPD susceptibility, and that an autoimmune mechanism may be implicated in the pathogenesis of the disease.
Maternal Exposure to the Holocaust and Health Complaints in Offspring
Authors: Flory Janine D. , Bierer Linda M. , Yehuda Rachel
Published: June 2013

Although the link between chronic stress and the development of cardiovascular and metabolic diseases of adulthood has been known for some time, there is growing recognition that early environmental influences may result in developmental programming via epigenetic mechanisms, thereby affecting the developmental trajectory of disease progression. Previous studies support the idea that offspring of Holocaust survivors may have been subjected to early developmental programming. We evaluated the relationship between parental exposure to the Holocaust and self-reported health ratings and disorders made by their adult offspring (i.e., second generation Holocaust survivors). A total of 137 subjects were evaluated. Regression analyses demonstrated that maternal but not paternal exposure to the Holocaust was related to poorer subjective impressions of emotional and physical health. This relationship was diminished when the offspring’s own level of trait anxiety was considered. Offspring with maternal, but not paternal, Holocaust exposure also reported greater use of psychotropic and other medications, including medications for the treatment of hypertension and lipid disorders. The mechanism linking these health outcomes and maternal exposure deserves further investigation, including the possibility that fetal or early developmental programming is involved.
Polymorphism in theKCNA3Gene Is Associated with Susceptibility to Autoimmune Pancreatitis in the Japanese Population
Authors: Ota Masao , Ito Tetsuya , Umemura Takeji , Katsuyama Yoshihiko , Yoshizawa Kaname , Hamano Hideaki , Kawa Shigeyuki
Published: June 2013

Autoimmune pancreatitis (AIP), characterized by irregular narrowing of the main pancreatic duct, swelling of the pancreas, and histological evidence of lymphoplasmacytic inflammation by high serum immunoglobulin G4, is distinct from ordinary pancreatitis. However, genetic factors involved in the etiology and pathophysiology of AIP remain unclear. Sixty-four patients with autoimmune pancreatitis (53 men, 11 women; mean age, 62.4 years) and 104 healthy Japanese controls were enrolled in this study. We performed an association analysis using 400 microsatellite markers with an average spacing of 10.8 cM in the genome. We also evaluated the association of AIP with seven single nucleotide polymorphisms (SNPs) within the 20-kb region around the potassium voltage-gated channel, shaker-related subfamily, member 3 gene (KCNA3). We identified six statistically significant markers (D1S2726, D5S410, D6S460, D10S548, D15S128, and D20S186;P< 0.05) related to susceptibility. The surrounding region showing the strong association (P= 7.4 × 10−7, Pc = 0.0015) contained theKCNA3gene. Further analysis by SNP genotyping inKCNA3gene revealed that four SNPs (rs2840381, rs1058184, rs2640480, rs1319782) were significantly associated with the AIP susceptibility (P< 0.007).KCNA3is known to be involved in immunomodulation of autoreactive effector and memory T cell–mediated autoimmune diseases. Our findings provide the first evidence thatKCNA3is associated with AIP and suggest thatKCNA3may influence the risk for AIP.
α-Defensin 1-3 And α-Defensin 4 as Predictive Markers of Imatinib Resistance and Relapse in CML Patients
Authors: Etienne Gabriel , Dupouy Maryse , Costaglioli Patricia , Chollet Claudine , Lagarde Valérie , Pasquet Jean-Max , Reiffers Josy , Garbay Bertrand , Mahon François-Xavier , Turcq Béatrice
Published: June 2013

Objective: Imatinib mesylate is a tyrosine kinase inhibitor used as first line treatment in chronic myeloid leukaemia. Despite a remarkable effectiveness, treatment failure cases have been reported in 20 percent of CML patients. The identification of biomarkers which can predict the response to imatinib is our point of interest.

Methods: Gene expression profiling microarray was carried out on secondary imatinib resistant patients. Longitudinal studies were performed on imatinib treated responder/resistant patients. Then, Q-RT/PCR studies were realized on patients prior imatinib initiation.

Results: For imatinib responder patients, we observed a strong and lasting decrease of α-defensin 1-3 and α-defensin 4 expression. For relapse patients, we observed a dramatic increase of α-defensin 1-3 and α-defensin 4 expression before BCR-ABL transcript increase. Moreover, before imatinib initiation, α-defensin 1-3 and α-defensin 4 expression was significantly lower in the resistant group than in the responder group.

ConclusionThe variation of expression of α-defensin 1-3 and α-defensin 4 in peripheral blood is associated with imatinib resistance and may reflect an adequate immune control of the disease. Monitoring of α-defensin 1-3 and α-defensin 4 could be helpful to predict the patients who are not going to respond to the treatment.
Elevated Levels of miR-146a and miR-155 in Kidney Biopsy and Urine from Patients with IgA Nephropathy
Authors: Wang Gang , Kwan Bonnie Ching-Ha , Lai Fernand Mac-Moune , Chow Kai-Ming , Li Philip Kam-Tao , Szeto Cheuk-Chun
Published: June 2013

Background: Previous studies suggested miR-146a and miR-155 play important roles in innate and adaptive immune responses. We studied intra-renal and urinary levels of miR-146a and miR-155 in patients with immunoglobulin A nephropathy (IgAN).

Methods:Intra-renal and urinary levels of miR-146a and miR-155 are quantified in 43 patients with IgAN; the result was compared to 20 nephrectomy specimens and urine sediment of 13 healthy volunteers.

Results: The levels of intra-renal and urinary levels of miR-146a and miR-155 of IgAN are significantly higher than controls. Estimated glomerular filtration rate inversely correlates with intra-renal level of miR-146a and miR-155; proteinuria positively correlates with intra-renal level of miR-146a and miR-155, as well as urinary level of miR-146a and miR-155. Intra-renal level of miR-155 significantly correlates with tubulointerstitial scarring. Urinary level of miR-146a inversely correlates with urinary expression of interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α and positively correlates with urinary expression of regulated upon activation, normal T-cell expressed, and secreted (RANTES). Urinary level of miR-155 inversely correlates with urinary expression of IL-1βand TNF-α and positively correlates with urinary expression of forkhead box P3 (FOXP3) and RANTES.

Conclusion:We conclude that intra-renal and urinary levels of miR-146a and miR-155 were significantly elevated in IgAN, and the degree of upregulation correlates with clinical and histological severity of the disease. Our results suggested miR-146a and miR-155 might play an important role in the pathophysiology of IgAN.
A Simple Oligonucleotide Biochip Capable of Rapidly Detecting Known Mitochondrial DNA Mutations in Chinese Patients with Leber’S Hereditary Optic Neuropathy (LHON)
Authors: Du Wei-Dong , Chen Gang , Cao Hui-Min , Jin Qing-Hui , Liao Rong-Feng , He Xiang-Cheng , Chen Da-Ben , Huang Shu-Ren , Zhao Hui , Lv Yong-Mei , Tang Hua-Yang , Tang Xian-Fa , Wang Yong-Qing , Sun Song , Zhao Jian-Long , Zhang Xue-Jun
Published: June 2013

Leber's hereditary optic neuropathy (LHON) is a maternally transmitted disease. Clinically, no efficient assay protocols have been available. In this study, we aimed to develop an oligonucleotide biochip specialized for detection of known base substitution mutations in mitochondrial DNA causing LHON and to investigate frequencies of LHON relevant variants in Anhui region of China. Thirty-two pairs of oligonucleotide probes matched with the mutations potentially linked to LHON were covalently immobilized. Cy5-lablled targets were amplified from blood DNA samples by a multiplex PCR method. Two kinds of primary mutations 11778 G > A and 14484 T > C from six confirmed LHON patients were interrogated to validate this biochip format. Further, fourteen Chinese LHON pedigrees and twenty-five unrelated healthy individuals were investigated by the LHON biochip, direct sequencing and pyrosequencing, respectively. The biochip was found to be able efficiently to discriminate homoplasmic and heteroplasmic mtDNA mutations in LHON. Biochip analysis revealed that twelve of eighteen LHON symptomatic cases from the 14 Chinese pedigree harbored the mutations either 11778G > A, 14484T > C or 3460G > A, respectively, accounting for 66.7%. The mutation 11778G > A in these patients was homoplasmic and prevalent (55.5%, 10 of 18 cases). The mutations 3460G > A and 3394T > C were found to co-exist in one LHON case. The mutation 13708G > A appeared in one LHON pedigree. Smaller amount of sampling and reaction volume, easier target preparation, fast and high-throughput were the main advantages of the biochip over direct DNA sequencing and pyrosequencing. Our findings suggested that primary mutations of 11778G > A, 14484T > C or 3460G > A are main variants of mtDNA gene leading to LHON in China. The biochip would easily be implemented in clinical diagnosis.
Potential Serological Biomarkers of Cerebral Malaria
Authors: Lucchi Naomi W. , Jain Vidhan , Wilson Nana O. , Singh Neeru , Udhayakumar Venkatachalam , Stiles Jonathan K.
Published: June 2013

Biomarkers have been used to diagnose and prognosticate the progress and outcome of many chronic diseases such as neoplastic and non communicable diseases. However, only recently did the field of malaria research move in the direction of actively identifying biomarkers that can accurately discriminate the severe forms of malaria. Malaria continues to be a deadly disease, killing close to a million people (mostly children) every year. One life-threatening complication of malaria is cerebral malaria (CM). Studies carried out in Africa have demonstrated that even with the best treatment, as high as 15–30% of CM patients die and about 10–24% of CM survivors suffer short-or long-term neurological impairment. The transition from mild malaria to CM can be sudden and requires immediate intervention. Currently, there is no biological test available to confirm the diagnosis of CM and its complications. It is hoped that development of biomarkers to identify CM patients and potential risk for adverse outcomes would greatly enhance better intervention and clinical management to improve the outcomes. We review here what is currently known regarding biomarkers for CM outcomes.

A Pub Med literature search was performed using the following search terms: “malaria,” “cerebral malaria,” “biomarkers,” “mortality” and “neurological sequelae.” This search revealed a paucity of usable biomarkers for CM management. We propose three main areas in which researchers can attempt to identify CM biomarkers: 1) early biomarkers, 2) diagnostic biomarkers and 3) prognostic biomarkers.
Combined Effect of ADH1B RS1229984, RS2066702 and ADH1C RS1693482/ RS698 Alleles on Alcoholism and Chronic Liver Diseases
Authors: Tóth Réka , Fiatal Szilvia , Petrovski Beáta , McKee Martin , Ádány Róza
Published: June 2013

The aim of this study was to analyze the combined effect of the most frequent alcohol dehydrogenase polymorphisms (Arg48His and Arg370Cys in ADH1B, Arg272Gln and Ile350Val in ADH1C) on the alcohol use habits, alcohol dependence and chronic liver diseases in Hungary.

The study included men, aged 45–64 years. Altogether, 241 cases with chronic liver disease (CLD) and 666 randomly selected controls without CLD were analysed for all four polymorphisms. Associations between the polymorphisms, individually, and in combination, and excessive and problem drinking and CLD, were assessed using logistic regression.

In this study we have identified a novel mutation, called ADH1B Arg370His. The ADH1C Arg272Gln and Ile350Val showed almost complete linkage. The 272Gln/350Val allele increased the risk of excessive and problem drinking in homozygous form (OR = 1.582,p= 0.035, CI = 1.034–2.421, OR = 1.780,p= 0.016, CI = 1.113–2.848, respectively). The joint analysis showed that when combined with the wild type ADH1C Arg272/Ile350 allele, the ADH1B 48His is protective against CLD (OR = 0.368,p= 0.019, CI = 0.159–0.851).

The results obtained in the study help not only to clarify the effects of different ADH SNPs but to better understand how these polymorphisms modify each other’s effects in the development of alcoholism and related diseases.
Detection of Deafness-Causing Mutations in the Greek Mitochondrial Genome
Authors: Kokotas Haris , Grigoriadou Maria , Korres George S. , Ferekidou Elisabeth , Kandiloros Dimitrios , Korres Stavros , Petersen Michael B.
Published: June 2013

Mitochondrion harbors its own DNA, known as mtDNA, encoding certain essential components of the mitochondrial respiratory chain and protein synthesis apparatus. mtDNA mutations have an impact on cellular ATP production and many of them are undoubtedly a factor that contributes to sensorineural deafness, including both syndromic and non-syndromic forms. Hot spot regions for deafness mutations are theMTRNR1gene, encoding the 12S rRNA, theMTTS1gene, encoding the tRNA for Ser(UCN), and theMTTL1gene, encoding the tRNA for Leu(UUR). We investigated the impact of mtDNA mutations in the Greek hearing impaired population, by testing a cohort of 513 patients suffering from childhood onset prelingual or postlingual, bilateral, sensorineural, syndromic or non-syndromic hearing loss of any degree for six mitochondrial variants previously associated with deafness. Screening involved theMTRNR1961delT/insC and A1555G mutations, theMTTL1A3243G mutation, and theMTTS1A7445G, 7472insC and T7510C mutations. Although two patients were tested positive for the A1555G mutation, we failed to identify any subject carrying the 961delT/insC, A3243G, A7445G, 7472insC, or T7510C mutations. Our findings strongly support our previously raised conclusion that mtDNA mutations are not a major risk factor for sensorineural deafness in the Greek population.
Tumor Necrosis Factor-Alpha Gene Promoter −308G/A and −238G/A Polymorphisms in Mexican Patients with Type 2 Diabetes Mellitus
Authors: Guzmán-Flores Juan Manuel , Muñoz-Valle José Francisco , Sánchez-Corona José , Cobián José G. , Medina-Carrillo Leopoldo , García-Zapién Alejandra G. , Cruz-Quevedo Edhit G. , Flores-Martínez Silvia Esperanza
Published: June 2013

The association between some Tumor necrosis factor-alpha (TNF-α) promoter polymorphisms and Type 2 diabetes mellitus (T2DM) remains controversial. Ethnic differences may play a role in these conflicting results. The aim of this study was to investigate the association between −308G/A and −238G/A polymorphisms located in the promoter region of the TNF-αgene and T2DM in Mexican mestizo patients. Nine hundred four individuals (259 patients with T2DM and 645 controls) were genotyped for the −308G/A and −238G/A polymorphisms by PCR—RFLP. We found that the −238A allele increased the risk of developing T2DM in Mexican patients (OR = 1.57, 95% CI: 1.07–2.29;p= 0.018). Moreover, we found that the frequency of the GA haplotype (created by the −308G and −238A alleles) was significantly increased in patients with T2DM when compared with controls (OR = 1.56, 95% CI: 1.05–2.31;p= 0.026). Our results suggest that the −238G/A polymorphism and a specific haplotype (GA) are genetic risk factors for the development of T2DM in Mexican population.
Gene Expression and Methylation Signatures ofMAN2C1are Associated with PTSD
Authors: Uddin Monica , Galea Sandro , Chang Shun-Chiao , Aiello Allison E. , Wildman Derek E. , de los Santos Regina , Koenen Karestan C.
Published: June 2013

As potential regulators of DNA accessibility and activity, epigenetic modifications offer a mechanism by which the environment can moderate the effects of genes. To date, however, there have been relatively few studies assessing epigenetic modifications associated with post-traumatic stress disorder (PTSD). Here we investigate PTSD-associated methylation differences in 33 genes previously shown to differ in whole blood-derived gene expression levels between those with vs. without the disorder. Drawing on DNA samples similarly obtained from whole blood in 100 individuals, 23 with and 77 without lifetime PTSD, we used methylation microarray data to assess whether these 33 candidate genes showed epigenetic signatures indicative of increased risk for, or resilience to, PTSD. Logistic regression analyses were performed to assess the main and interacting effects of candidate genes’ methylation values and number of potentially traumatic events (PTEs), adjusting for age and other covariates. Results revealed that only one candidate gene–MAN2C1–showed a significant methylation x PTE interaction, such that those with both higher MAN2C1 methylation and greater exposure to PTEs showed a marked increase in risk of lifetime PTSD (OR 4.35, 95% CI: 1.07, 17.77,p= 0.04). These results indicate that MAN2C1 methylation levels modify cumulative traumatic burden on risk of PTSD, and suggest that both gene expression and epigenetic changes at specific loci are associated with this disorder.
Platelet-Stored Angiogenesis Factors: Clinical Monitoring Is Prone to Artifacts
Authors: Starlinger Patrick , Alidzanovic Lejla , Schauer Dominic , Brugger Philipp , Sommerfeldt Silvia , Kuehrer Irene , Schoppmann Sebastian F. , Gnant Michael , Brostjan Christine
Published: June 2013

Background:The analysis of angiogenesis factors in the blood of tumor patients has given diverse results on their prognostic or predictive value. Since mediators of angiogenesis are stored in platelets, their measurement in plasma is sensitive to inadvertent platelet activation during blood processing.

Methods:Variants of blood withdrawal and plasma preparation were evaluated by ELISA for the detection of TSP-1, PF-4, VEGF and PD-ECGF. A total of 22 pancreatic cancer patients and 29 healthy volunteers were evaluated.

Results:Plasma preparation with the anticoagulant mix of citrate, theophylline, adenosine, dipyridamole (CTAD) and immediate blood processing at 4°C was required for reproducible measurements of TSP-1, PF-4 and VEGF. Blood collection by venflon or inadvertent hemolysis during blood withdrawal caused significantly elevated TSP-1 and PF4 values. When optimized plasma preparation was applied, a significant increase of TSP-1 and VEGF in cancer patients was detected (P = 0.006; P < 0.001).

Conclusion:The reliable plasma analysis of circulating platelet-stored angiogenesis factors requires preparation with CTAD at 4°C and blood collection by butterfly needle. Suboptimal procedures of plasma preparation are commonly applied in clinical monitoring of angiogenesis parameters which may account for the differences in reported plasma values and may have masked their predictive or prognostic marker potential.
Transcriptional Control of Monocyte Gene Expression in Post-Traumatic Stress Disorder
Authors: O’Donovan Aoife , Sun Bing , Cole Steve , Rempel Hans , Lenoci Maryann , Pulliam Lynn , Neylan Thomas
Published: June 2013

Post-traumatic stress disorder (PTSD) confers an increased risk for disorders with an inflammatory etiology. PTSD-related dysregulation of the sympathetic nervous system (SNS) and hypothalamic-pituitary adrenal (HPA) axis and associated alterations in inflammatory activity may contribute to this increased risk. However, little is known about convergent SNS, HPA and inflammatory signaling at the level of the immune cell transcriptome in PTSD. To explore such signaling, we examined the prevalence of specific transcription factor binding motifs in the promoter regions of differentially expressed genes in monocytes from individuals with PTSD and matched controls. Participants included 49 men (24 PTSD+ and 25 trauma-exposed controls) and 18 women (10 PTSD+ and 8 controls). Men with PTSD showed up-regulation of target genes for the NF-κB/Rel family of transcription factors, which convey inflammatory signals, up-regulation of target genes for CREB/ATF transcription factors, which convey adrenergic signals from the SNS, and down-regulation of target genes for the glucocorticoid receptor, which conveys glucocorticoid signals from the HPA axis. Women with PTSD also showed significant up-regulation of target genes for NF-κB and non-significant down-regulation of target genes for GR, but significant down-regulation of target genes for CREB/ATF. Altered transcriptional control of monocyte gene expression could contribute to exaggerated inflammatory activity in PTSD.
HFE Gene Polymorphisms and the Risk for Autism in Egyptian Children and Impact on the Effect of Oxidative Stress
Authors: Gebril Ola H. , Meguid Nagwa A.
Published: June 2013

Background: Autism is among the commonest neurodevelopmental childhood disorders worldwide; its aetiology is still unknown. Iron metabolism alteration in the central nervous system is recently implicated as a risk factor for several neurodegenerative disorders.

Haemochromatosis HFE gene polymorphisms (p.H63D and p.C282Y) have shown significant association with several neurological diseases. Some evidences show altered iron related proteins in serum of autistic children. The aim of this work is to conduct a preliminary pilot study for the association of HFE polymorphisms and autism.

Methods: All cases were referred from the clinic of special needs, National Research Centre, Cairo. Clinical diagnosis was based on the criteria for autistic disorder as defined in the Diagnostic and Statistical Manual of Mental Disorders Fourth Edition, Text Revision (DSM-IV-TR).

Whole genome DNA was extracted; p.H63D and p.C282Y genotyping was studied using specific sequence amplification followed by restriction enzyme digestion on a sample of autism patients (25 cases) and twenty controls.

Results: The p.H63D is more abundant than the C282Y among both autism and control samples. No significant association of p.H63D nor p.C282Y polymorphism and autism was revealed.

Conclusion: We here report on the first pilot study of the possible genetic association between autism and HFE gene polymorphisms among Egyptians. Although our results do not prove the role of HFE polymorphisms as risk factors for autism, yet this does not exclude the role of iron in this prevalent disorder. Further extended studies are recommended to include other iron metabolism genes.
Use of Host- and Bacteria-Derived Salivary Markers in Detection of Periodontitis: A Cumulative Approach
Authors: Gursoy Ulvi Kahraman , Könönen Eija , Pussinen Pirkko J. , Tervahartiala Taina , Hyvärinen Kati , Suominen Anna L. , Uitto Veli-Jukka , Paju Susanna , Sorsa Timo
Published: June 2013

In the present study, we propose a novel diagnostic approach, using 3 different salivary markers, representing periodontal pathogen burden, inflammation, and tissue degradation, for detecting periodontitis. The salivary concentrations ofPorphyromonas gingivalis, interleukin-1β, and matrix metalloproteinase-8, available from salivary specimens of 165 subjects (84 subjects with advanced periodontitis and 81 controls), were calculated together to obtain a cumulative risk score (CRS). In the calculation of CRS, the concentrations of each marker were divided into tertiles, and cumulative sub-score per each subject were calculated by the multiplication of the tertile values. Three CRS groups, indicating the lowest, medium, or highest risk, were formed with the cumulative sub-scores. Logistic regression analysis and ROC curves were performed to study the association of CRS with periodontitis. The results indicate that CRS, calculated from the 3 salivary biomarkers, is associated with advanced periodontitis more strongly than any of the markers individually. CRS offers a novel, non-invasive model for advanced periodontitis risk categorization that is especially useful in large population surveys where a periodontal examination is not feasible.
Association ofGSTM1andGSTT1Polymorphism with Lipid Peroxidation in Benign Prostate Hyperplasia and Prostate Cancer: A Pilot Study
Authors: Kumar Vivek , Yadav Chandra Shekhar , Datta Sudip Kumar , Singh Satyender , Ahmed Rafat S. , Goel Sanjay , Gupta Sanjay , Mustafa Md. , Grover Rajesh Kumar , Banerjee Basu Dev
Published: June 2013

Association of glutathione S-transferase (GST)M1andT1deletions with benign prostate hyperplasia (BPH) and prostate cancer is well reported. These enzymes metabolize numerous toxins thus protecting from oxidative injury. Oxidative stress has been associated with development of BPH and prostate cancer. The present study was designed to analyze role ofGSTdeletions in development of oxidative stress in these subjects. GSTs are responsible for metabolism of toxins present in tobacco therefore effect of tobacco usage in study groups was also studied. Three groups of subjects: BPH (57 patients), prostate cancer (53 patients) and controls (46 subjects) were recruited. Genotyping was done using a multiplex polymerase chain reaction (PCR) method. Malondialdehyde (MDA) levels as marker of oxidative stress were estimated by measuring thiobarbituric acid reactive substance (TBARS) in plasma. Based on genotyping, subjects were categorized into:GSTM1+/GSTT1+, GSTM1-/GSTT1+, GSTM1+/GSTT1- and GSTM1-/GSTT1-. Significantly higher plasma MDA levels were noticed inGSTM1-/GSTT1- as compared toGSTM1+/GSTT1+in all study groups. Double deletion (GSTM1-/GSTT1-) is associated with higher oxidative stress which might play a role in the pathogenesis of BPH and prostate cancer. However, other markers of oxidative stress should be analyzed before any firm conclusion.
Plasma Oxidized Low-Density Lipoprotein Is an Independent Risk Factor in Young Patients with Coronary Artery Disease
Authors: Huang Yuli , Hu Yunzhao , Mai Weiyi , Cai Xiaoyan , Song Yuanbin , Wu Yanxian , Dong Yugang , Huang Huiling , He Zhongyun , Li Wensheng , Yang You , Rao Shaoqi
Published: June 2013

Objectives:Oxidized low-density lipoprotein (ox-LDL) is considered to be a key factor of initiating and accelerating atherosclerosis. The objective of this study was to investigate the role of ox-LDL in young patients with coronary artery disease (CAD).

Methods:128 consecutive angiographically proven young CAD patients (aged ≤ 55 years) were enrolled, and 132 age-matched non-CAD individuals (coronary angiography normal or negative finding by coronary ultrafast CT) were set as control group. Conventional risk factors (hypertension, dyslipidemia, diabetes mellitus, obesity, smoking) were evaluated in the two groups. Ox-LDL was measured by competitive ELISA. Framingham risk score (FRS) and absolute 10-year CAD events risk were calculated for each individual.

Results:Male sex was more prevalent in group CAD than in control (87.5% vs. 62.1%;P< 0.01). There were significant differences in smoking history (P< 0.01) and triglyeride (TG) and ratio of apolipoprotein B/apolipoprotein A1 (ApoB/ApoA1) (bothP< 0.05) but no remarkable difference in other conventional risk factors (allP> 0.05) between group CAD and control. Level of ox-LDL was significantly higher in group CAD than in control (P < 0.01). Multivariate logistic regression showed that male sex (OR, 4.54; 95%CI, 1.76–9.77), smoking quantity (OR, 2.78; 95%CI, 1.34–4.25), TG (OR, 1.42; 95%CI, 1.18–2.83), ApoB/ApoA1 (OR, 1.73; 95%CI, 1.32–4.23), and ox-LDL (OR, 2.15; 95%CI, 1.37–6.95) were independently correlated with CAD in young patients. Area under the curve (AUC) of receiver operating characteristic (ROC) curve of TG, ApoB/ApoA1, and ox-LDL was 0.831, 0.866, and 0.935, respectively (P< 0.001).

Conclusions:Ox-LDL is an important independent risk factor for CAD in young patients after adjusting other risk factors such as smoking, TG, and ApoB/ApoA1.
MMP-9, Homocysteine and CRP Circulating Levels Are Associated with Intraluminal Thrombus Thickness of Abdominal Aortic Aneurysms–New Implication of the Old Biomarkers
Authors: Wiernicki Ireneusz , Millo Barbara , Safranow Krzysztof , Gorecka-Szyld Barbara , Gutowski Piotr
Published: June 2013

Background:Abdominal aortic aneurysms (AAAs) are characterized by presence of high proteolytic activity, atherosclerotic lesions, extensive transmural inflammation and the presence of variably sized and shaped intraluminal thrombus (ILT). Therefore, we evaluated a possible association between plasma matrix metalloproteinase-9 (MMP-9), homocysteine (Hcy), high-sensitivity C-reactive protein (hsCRP) levels and ILT thickness in patients with AAA.

Methods:Plasma concentrations of MMP-9, Hcy and hsCRP were determined and ILT thickness was measured in 71 patients with AAA. They were divided into 2 groups according to ILT thickness: 34 patients with ILT mean thickness ≥ 9 mm and 37 patients with ILT < 9 mm.

Results:Plasma MMP-9 and CRP concentrations in patients with thin ILT were significantly higher than in group with thick ILT (medians 610 vs. 485 ng/mL,p= 0.00003, and 7.7 vs. 3.3 mg/L,p< 0.00001, respectively) In contrast, plasma Hcy concentrations in patients with thin ILT were significantly lower than in the group with thick ILT (medians 14.3 vs. 19.2 μmol/L,p< 0.00001). Multiple regression models adjusted for age and AAA diameter showed that thin ILT is an independent predictor of high MMP-9 and CRP concentrations, while thick ILT predicts high Hcy concentrations.

Conclusions:Association of higher plasma levels of MMP-9 and CRP with thin ILT may be related to two phenomena: thin thrombi convey more elastolysis-stimulating factors from blood to the AAA wall and thin thrombi convey more factors involved in proteolysis and inflammation from AAA wall to blood. The association of thin ILT with lower plasma Hcy concentrations may be related to the role of Hcy as a prothrombotic marker and needs further research.
Mcl-1 Expression Is Up-Regulated in Malignancies of the Parotid Gland
Authors: Heiduschka Gregor , Erovic Boban M. , Pammer Johannes , Kotowski Ulana , Kaider Alexandra , Ch. Grasl Matthaeus , Thurnher Dietmar
Published: June 2013

The anti-apoptotic protein Mcl-1 is highly expressed in various types of malignant tumors. Overexpression is reported to correlate with poor prognosis and disease progression. We report the expression levels of Mcl-1 in tumor samples of the parotid gland. A retrospective study containing 108 patients was performed. A tissue microarray of six malignancies of the parotid gland and pleomorphic adenoma as control was constructed. Parotid gland tumor samples were immunohistochemically stained for Mcl-1 and expression intensities were assessed. Statistical analysis included correlation to patients' clinical data and comparison of malignancies to the adenoma.

All malignancies had significantly higher expression of Mcl-1 than the pleomorphic adenomas. The intensity, however, had no significant correlation to overall survival.

Our immunohistochemical findings indicate that parotid gland malignancies produce high levels of Mcl-1 protein. Therefore, Mcl-1 might serve as a predictive co-marker in tumors of the parotid gland.