bluuebunny/crossref_metadata_2025 huggingface dataset:
Urinary exosomal Wilms' tumor-1 as a potential biomarker for podocyte injury
Authors: Zhou Hua , Kajiyama Hiroshi , Tsuji Takayuki , Hu Xuzhen , Leelahavanichkul Asada , Vento Suzanne , Frank Rachel , Kopp Jeffrey B. , Trachtman Howard , Star Robert A. , Yuen Peter S. T.
Published: June 2013

Renal Wilms' tumor-1 (WT-1) staining is used to detect podocyte loss in kidney biopsies. We aimed to determine if urinary exosomal WT-1 could serve as a noninvasive biomarker of podocyte injury. We examined WT-1 by Western blot in a human podocyte-like cell line, a mouse model of podocyte injury, and human subjects with podocyte disorders. WT-1 was detected in exosomal fraction of the conditioned media from podocytes and increased 48 h after hTGF-β1 stimulation. Cellular WT-1 decreased in podocytes following hTGF-β1 incubation. In mice with induced podocyte injury, urinary exosomal WT-1 was detected 1 wk earlier than albuminuria and also tracked the effects of angiotensin receptor blocker (ARB) treatment. In addition, urinary exosomal WT-1 levels at 1 wk post-injury correlated with the severity of glomerular injury at 3 wk later. In human subjects, urinary exosomal WT-1 was significantly increased in focal segmental glomerulosclerosis (FSGS) patients compared with healthy volunteers or steroid-sensitive nephrotic syndrome (SSNS) patients. Urinary exosomal WT-1 was also significantly decreased in patients in remission for either FSGS or SSNS or following steroid treatment in six SSNS subjects. We conclude that urinary exosomal WT-1 is a promising noninvasive biomarker with apparent podocyte specificity that can detect early progression and treatment-induced regression of podocyte injury in FSGS or SSNS. These results warrant longitudinal, prospective studies in a large cohort with a range of podocyte diseases.
Myosin 1e is a component of the glomerular slit diaphragm complex that regulates actin reorganization during cell-cell contact formation in podocytes
Authors: Bi J. , Chase S. E. , Pellenz C. D. , Kurihara H. , Fanning A. S. , Krendel M.
Published: June 2013

Glomerular visceral epithelial cells, also known as podocytes, are critical to both normal kidney function and the development of kidney disease. Podocyte actin cytoskeleton and their highly specialized cell-cell junctions (also called slit diaphragm complexes) play key roles in controlling glomerular filtration. Myosin 1e (myo1e) is an actin-based molecular motor that is expressed in renal glomeruli. Disruption of the Myo1e gene in mice and humans promotes podocyte injury and results in the loss of the integrity of the glomerular filtration barrier. Here, we have used biochemical and microscopic approaches to determine whether myo1e is associated with the slit diaphragm complexes in glomerular podocytes. Myo1e was consistently enriched in the slit diaphragm fraction during subcellular fractionation of renal glomeruli and colocalized with the slit diaphragm markers in mouse kidney. Live cell imaging studies showed that myo1e was recruited to the newly formed cell-cell junctions in cultured podocytes, where it colocalized with the actin filament cables aligned with the nascent contacts. Myo1e-null podocytes expressing FSGS-associated myo1e mutant (A159P) did not efficiently assemble actin cables along new cell-cell junctions. We have mapped domains in myo1e that were critical for its localization to cell-cell junctions and determined that the SH3 domain of myo1e tail interacts with ZO-1, a component of the slit diaphragm complex and tight junctions. These findings suggest that myo1e represents a component of the slit diaphragm complex and may contribute to regulating junctional integrity in kidney podocytes.
Neuromodulation of motor-evoked potentials during stepping in spinal rats
Authors: Gad Parag , Lavrov Igor , Shah Prithvi , Zhong Hui , Roy Roland R. , Edgerton V. Reggie , Gerasimenko Yury
Published: June 2013

The rat spinal cord isolated from supraspinal control via a complete low- to midthoracic spinal cord transection produces locomotor-like patterns in the hindlimbs when facilitated pharmacologically and/or by epidural electrical stimulation. To evaluate the role of epidural electrical stimulation in enabling motor control (eEmc) for locomotion and posture, we recorded potentials evoked by epidural spinal cord stimulation in selected hindlimb muscles during stepping and standing in adult spinal rats. We hypothesized that the temporal details of the phase-dependent modulation of these evoked potentials in selected hindlimb muscles while performing a motor task in the unanesthetized state would be predictive of the potential of the spinal circuitries to generate stepping. To test this hypothesis, we characterized soleus and tibialis anterior (TA) muscle responses as middle response (MR; 4–6 ms) or late responses (LRs; >7 ms) during stepping with eEmc. We then compared these responses to the stepping parameters with and without a serotoninergic agonist (quipazine) or a glycinergic blocker (strychnine). Quipazine inhibited the MRs induced by eEmc during nonweight-bearing standing but facilitated locomotion and increased the amplitude and number of LRs induced by eEmc during stepping. Strychnine facilitated stepping and reorganized the LRs pattern in the soleus. The LRs in the TA remained relatively stable at varying loads and speeds during locomotion, whereas the LRs in the soleus were strongly modulated by both of these variables. These data suggest that LRs facilitated electrically and/or pharmacologically are not time-locked to the stimulation pulse but are highly correlated to the stepping patterns of spinal rats.
Temporal changes of neocortical high-frequency oscillations in epilepsy
Authors: Pearce Allison , Wulsin Drausin , Blanco Justin A. , Krieger Abba , Litt Brian , Stacey William C.
Published: June 2013

High-frequency (100–500 Hz) oscillations (HFOs) recorded from intracranial electrodes are a potential biomarker for epileptogenic brain. HFOs are commonly categorized as ripples (100–250 Hz) or fast ripples (250–500 Hz), and a third class of mixed frequency events has also been identified. We hypothesize that temporal changes in HFOs may identify periods of increased the likelihood of seizure onset. HFOs (86,151) from five patients with neocortical epilepsy implanted with hybrid (micro + macro) intracranial electrodes were detected using a previously validated automated algorithm run over all channels of each patient's entire recording. HFOs were characterized by extracting quantitative morphologic features and divided into four time epochs (interictal, preictal, ictal, and postictal) and three HFO clusters (ripples, fast ripples, and mixed events). We used supervised classification and nonparametric statistical tests to explore quantitative changes in HFO features before, during, and after seizures. We also analyzed temporal changes in the rates and proportions of events from each HFO cluster during these periods. We observed patient-specific changes in HFO morphology linked to fluctuation in the relative rates of ripples, fast ripples, and mixed frequency events. These changes in relative rate occurred in pre- and postictal periods up to thirty min before and after seizures. We also found evidence that the distribution of HFOs during these different time periods varied greatly between individual patients. These results suggest that temporal analysis of HFO features has potential for designing custom seizure prediction algorithms and for exploring the relationship between HFOs and seizure generation.
Isolation and high-throughput sequencing of two closely linked epistatic hypertension susceptibility loci with a panel of bicongenic strains
Authors: Pillai Resmi , Waghulde Harshal , Nie Ying , Gopalakrishnan Kathirvel , Kumarasamy Sivarajan , Farms Phyllis , Garrett Michael R. , Atanur Santosh S. , Maratou Klio , Aitman Timothy J. , Joe Bina
Published: June 2013

Interactions or epistasis between genetic factors may contribute to “missing heritability.” While linkage analyses detect epistasis, defining the limits of the interacting segments poses a significant challenge especially when the interactions are between loci in close proximity. The goal of the present study was to isolate two such epistatic blood pressure (BP) loci on rat chromosome 5. A panel of S.LEW bicongenic strains along with the corresponding monocongenic strains was constructed. BP of each set comprising of one bicongenic and two corresponding monocongenic strains were determined along with the parental Salt-sensitive (S) strain. Epistasis was observed in one out of four sets of congenic strains, wherein systolic blood pressures (SBP) of the two monocongenic strains S.LEW(5)x6Bx9x5a and S.LEW(5)x6Bx9x5b were comparable to that of S, but the SBP of the bicongenic strain S.LEW(5)x6Bx9x5 (157 ± 4.3 mmHg) was significantly lower than that of S (196 ± 6.8 mmHg, P < 0.001). A two-way ANOVA indicated significant interactions between the LEW alleles at the two loci. The interacting loci were 2.02 Mb apart and located within genomic segments spanning 7.77 and 4.18 Mb containing 7,360 and 2,753 candidate variants, respectively. The current study demonstrates definitive evidence for epistasis and provides genetic tools for further dissection of the isolated epistatic BP loci.
Transcriptional Activity of HTLV‐I Tax Influences the Expression of Marker Genes Associated with Cellular Transformation
Authors: Lemoine Francene J. , Wycuff Diane R. , Marriott Susan J.
Published: June 2013

Human T cell leukemia virus type I (HTLV‐I) has been identified as the etiologic agent of adult T cell leukemia (ATL). HTLV‐I encodes a transcriptional regulatory protein, Tax, which also functions as the viral transforming protein. Through interactions with a number of cellular transcription factors Tax can modulate cellular gene expression. Since the majority of Tax‐responsive cellular genes are important regulators of cellular proliferation, the transactivating functions of Tax appear to be necessary for cellular transformation by HTLV‐I. Gaining a complete understanding of the broad range of genes regulated by Tax, the temporal pattern of their expression, and their effects on cell function may identify early markers of disease progression mediated by this virus.
Evaluation of Blood Units with Isolated Anti HBC for the Presence of HBV DNA
Authors: Zekri Abdel-Rahman N. , Awlia Azza A. , Mahalawi Hadeer El , Ismail Emad F. , Mabrouk Gamal M.
Published: June 2013

We screened blood donors in one center in Saudi Arabia for a safety transfusion. We found that among 5043 blood donors negative for HCV and HIV, the incidence of HBsAg positivity was 2.97%. When antiHBc antibody was measured (HBcIg) in HBsAg negative donors, we observed that 21.47% were positive indicating previous exposure to hepatitis B virus. The HBcIg positive blood was further screened for HBsAb and the specimens were found to be reactive in 81.54%. Based on these data blood transfusion was permissible from donors who showed HBsAg negativity, HBcIg positive and HBsAb reactive blood. In order to ensure safety transfusion an aliquot of specimens (n = 80) was further analyzed for HBV DNA by PCR. We found only one specimen positive with incidence of 1.25%. So we recommended restricting transfusion from the previously mentioned donors to emergencies.
Surfactant Protein B Intron 4 Variation in German Patients with COPD and Acute Respiratory Failure
Authors: Seifart Carola , Plagens Alexandra , Brödje Dörte , Müller Bernd , von Wichert Peter , Floros Joanna
Published: June 2013

Chronic obstructive pulmonary disease (COPD) is a major health problem. Genetic factors that contribute to the disease have been postulated. The pulmonary surfactant protein B (SP‐B), which is essential for normal lung function, is considered as a candidate gene for COPD in this case‐control study. We studied the SP‐B intron 4 size variants in 346 individuals. This group consisted of 118 patients with chronic bronchitis or COPD, including 24 patients with acute respiratory failure (ARF) in COPD, 118 matched controls without pulmonary disease and 110 healthy individuals (population control). The frequency of intron 4 variants was similar in either control group (10.9%, 14.4% respectively), with a small increase in the COPD group (18.6%). This increase was due to a high increase of intron 4 variants in the ARF subgroup (37.5%, p = 0.003, OR 4.9, 95% CI: 1.76–13.6). The data indicate that SP‐B intron 4 variants may associate with increased risk of ARF in COPD and may be used as a marker of susceptibility in this disease subgroup.
The Stromelysin‐1 5A/6A Promoter Polymorphism Is a Disease Marker for the Extent of Coronary Heart Disease
Authors: Schwarz Adelheid , Haberbosch Werner , Tillmanns Harald , Gardemann Andreas
Published: June 2013

Background. Matrix metalloproteinases, such as stromelysin‐1, are implicated in the pathogenesis of coronary artery disease (CAD) and acute myocardial infarction (MI). A 5A/6A promoter polymorphism can regulate the transcription of the stromelysin‐1 gene in an allele‐specific manner. Evidence has been presented that the 6A allele is associated with the progression of coronary heart disease (CHD). In contrast, the 5A allele may be linked to the risk of MI.

Results. To analyse the relation of the 5A/6A polymorphism with the risk and severity of CHD and the risk of MI, a case‐control study of 515 healthy controls and 1848 participants who underwent coronary angiography for diagnostic purposes was conducted. In the total sample, the mean CHD scores—according to Gensini—were different between 5A/6A genotypes: 5A5A homozygotes had the lowest, 6A6A genotypes the highest and 5A6A heterozygotes intermediate scores. These differences were even more pronounced when the participants were restricted to individuals with a high coronary risk profile (high apoB levels, high Lp(a) levels, high glucose levels, combinations of either high apoB and Lp(a) levels or high apoB, Lp(a) and glucose plasma levels). Mean values were used as cut points for high‐risk populations, respectively. In contrast, the 5A allele was not associated with the risk of CHD or MI. Even when angiographically controlled individuals without MI were compared with MI patients in subpopulations of participants with no, single, double and triple vessel disease, the frequencies of the 5A/6A and/or the 5A5A genotypes were not higher in each subgroup, respectively.

Conclusions. The present results do not confirm an association of the 5A allele with the risk of MI, observed in another investigation, but strengthen the hypothesis of earlier studies that the 6A allele is a disease marker for progression of coronary heart disease. Further investigations should evaluate whether 6A allele carriers and especially 6A homozygotes might benefit from a more aggressive therapy against CHD progression.
Assessment of Cancer‐Associated Biomarkers by Positron Emission Tomography: Advances and Challenges
Authors: Collier T. Lee , Lecomte Roger , McCarthy Timothy J. , Meikle Steve , Ruth Thomas J. , Scopinaro Francesco , Signore Alberto , Van Brocklin Henry , Van de Wiele Christophe , Waterhouse Rikki N.
Published: June 2013

Positron emission tomography (PET) provides a powerful means to non‐invasively image and quantify protein expression and biochemical changes in living subjects at nano‐ and picomolar levels. As the field of molecular imaging develops, and as advances in the biochemistry, pharmacology, therapeutics, and molecular biology of disease are made, there is a corresponding increase in the number of clinically relevant, novel disease‐associated biomarkers that are brought to the attention of those developing imaging probes for PET. In addition, due to the high specificity of the PET radiotracers being developed, there is a demand for PET cameras with higher sensitivity and resolution. This manuscript reviews advances over the past five years in clinical and pre‐clinical PET instrumentation and in new PET probes and imaging methods associated with the latest trends in the molecular imaging of cancer. Included in the PET tracer review is a description of new radioligands for steroid receptors, growth factor receptors, receptor tyrosine kinases, sigma receptors, tumor‐associated enzymes, gene reporter probes, markers for tumor hypoxia and metabolism, and sites associated with angiogenesis and cellular proliferation. The use of PET imaging in drug development, including the monitoring of cancer chemotherapy, also is discussed.
Overview of Biomarkers and Surrogate Endpoints in Drug Development
Authors: Wagner John A.
Published: June 2013

There are numerous factors that recommend the use of biomarkers in drug development including the ability to provide a rational basis for selection of lead compounds, as an aid in determining or refining mechanism of action or pathophysiology, and the ability to work towards qualification and use of a biomarker as a surrogate endpoint. Examples of biomarkers come from many different means of clinical and laboratory measurement. Total cholesterol is an example of a clinically useful biomarker that was successfully qualified for use as a surrogate endpoint. Biomarkers require validation in most circumstances. Validation of biomarker assays is a necessary component to delivery of high‐quality research data necessary for effective use of biomarkers. Qualification is necessary for use of a biomarker as a surrogate endpoint. Putative biomarkers are typically identified because of a relationship to known or hypothetical steps in a pathophysiologic cascade. Biomarker discovery can also be effected by expression profiling experiment using a variety of array technologies and related methods. For example, expression profiling experiments enabled the discovery of adipocyte related complement protein of 30 kD (Acrp30 or adiponectin) as a biomarker for in vivo activation of peroxisome proliferator‐activated receptors (PPAR) γ activity.
Biomarker Amplification by Serum Carrier Protein Binding
Authors: Mehta Arpita I. , Ross Sally , Lowenthal Mark S. , Fusaro Vincent , Fishman David A. , Petricoin Emanuel F. , Liotta Lance A.
Published: June 2013

Mass spectroscopic analysis of the low molecular mass (LMM) range of the serum/plasma proteome is a rapidly emerging frontier for biomarker discovery. This study examined the proportion of LMM biomarkers, which are bound to circulating carrier proteins. Mass spectroscopic analysis of human serum following molecular mass fractionation, demonstrated that the majority of LMM biomarkers exist bound to carrier proteins. Moreover, the pattern of LMM biomarkers bound specifically to albumin is distinct from those bound to non‐albumin carriers. Prominent SELDI‐TOF ionic species (m/z 6631.7043) identified to correlate with the presence of ovarian cancer were amplified by albumin capture. Several insights emerged: a) Accumulation of LMM biomarkers on circulating carrier proteins greatly amplifies the total serum/plasma concentration of the measurable biomarker, b) The total serum/plasma biomarker concentration is largely determined by the carrier protein clearance rate, not the unbound biomarker clearance rate itself, and c) Examination of the LMM species bound to a specific carrier protein may contain important diagnostic information. These findings shift the focus of biomarker detection to the carrier protein and its biomarker content.
Development of a Fluorescent Multiplex Assay for Detection of MSI‐High Tumors
Authors: Bacher Jeffery W. , Flanagan Laura A. , Smalley Regenia L. , Nassif Nadine A. , Burgart Lawrence J. , Halberg Richard B. , Megid Wael M. Abdel , Thibodeau Stephen N.
Published: June 2013

Determining whether a tumor exhibits microsatellite instability (MSI) is useful in identifying patients with hereditary non‐polyposis colorectal cancer and sporadic gastrointestinal cancers with defective DNA mismatch repair (MMR). The assessment of MSI status aids in establishing a clinical prognosis and may be predictive of tumor response to chemotherapy. A reference panel of five markers was suggested for MSI analysis by a National Cancer Institute (NCI) workshop in 1997 that has helped to standardize testing. But this panel of markers has limitations resulting from the inclusion of dinucleotide markers, which are less sensitive and specific for detection of tumors with MMR deficiencies compared to other types of markers that are currently available. This study demonstrates that mononucleotides are the most sensitive and specific markers for detection of tumors with defects in MMR and identifies an optimal panel of markers for detection of MSI‐H tumors. A set of 266 mono‐, di‐, tetra‐ and penta‐nucleotide repeat microsatellite markers were used to screen for MSI in colorectal tumors. The best markers for detection of MSI‐H tumors were selected for a MSI Multiplex System, which included five mononucleotide markers: BAT-25, BAT-26, NR-21, NR-24 and MONO-27. In addition, two pentanucleotide markers were added to identify sample mix‐ups and/or contamination. We classified 153 colorectal tumors using the new MSI Multiplex System and compared the results to those obtained with a panel of 10 microsatellite markers combined with immunohistochemical (IHC) analysis. We observed 99% concordance between the two methods with nearly 100% accuracy in detection of MSI‐H tumors. Approximately 5% of the MSI‐H tumors had normal levels of four MMR proteins and as a result would have been misclassified based solely on IHC analysis, emphasizing the importance of performing MSI testing. The new MSI Multiplex System offers several distinct advantages over other methods of MSI testing in that it is both extremely sensitive and specific and amenable to high‐throughput analysis. The MSI Multiplex System meets the new recommendations proposed at the recent 2002 NCI workshop on HNPCC and MSI testing and overcomes problems inherent to the original five‐marker panel. The use of a single multiplex fluorescent MSI assay reduces the time and costs involved in MSI testing with increased reliability and accuracy and thus should facilitate widespread screening for microsatellite instability in tumors of patients with gastrointestinal cancers.
Development Tracks for Cancer Prevention Markers
Authors: Baker Stuart G. , Kramer Barnett S. , Prorok Philip C.
Published: June 2013

We provide a general framework for describing various roles for biomarkers in cancer prevention research (early detection, surrogate endpoint, and cohort identification for primary prevention) and the phases in their evaluation.
Colorectal Carcinogenesis: MSI‐H Versus MSI‐L
Authors: Pawlik Timothy M. , Raut Chandrajit P. , Rodriguez-Bigas Miguel A.
Published: June 2013

Microsatellite instability (MSI) is a well‐recognized phenomenon that is classically a feature of tumors in the hereditary non‐polyposis colorectal syndrome. Ten to 15% of sporadic colorectal cancers, however, will have MSI. Microsatellite unstable tumors can be divided into two distinct MSI phenotypes: MSI‐high (MSI‐H) and MSI‐low (MSI‐L). MSI sporadic colorectal cancers with a high level of MSI (MSI‐H) form a well defined group with distinct clinicopathologic features characterized by an overall better long‐term prognosis. These sporadic MSI‐H colorectal tumors most often arise from the epigenetic silencing of the mismatch repair gene MLH1. In contrast, MSI‐L colorectal tumors have not been shown to differ in their clinicopathologic features or in most molecular features from microsatellite stable (MSS) tumors. Unlike MSI‐H tumors, MSI‐L tumors appear to arise through the chromosomal instability carcinogenesis pathway, similar to MSS tumors. Some groups have reported more frequent mutations in K‐ras and in the methylation of methylguanine transferase in MSI‐L tumors, but others have questioned these findings. Therefore, although the use of the MSI‐L category is widespread, there continues to be some debate as to whether a discrete MSI‐L group truly exists. Rather, it has been suggested that MSI‐L tumors differ quantitatively from MSS tumors but do not differ qualitatively. Future studies will need to evaluate the specific mutations in non‐MSI‐H tumors in an attempt to sub‐classify MSI‐L tumors with regard to MSS tumors so that subtle differences between these two sub‐groups can be identified.
Update on Diabetes Mellitus
Authors: Korc Murray
Published: June 2013

Diabetes mellitus is a complex multi‐system disorder that may be classified as autoimmune mediated type 1 diabetes, or as insulin resistance associated type 2 diabetes. In type 1 diabetes, there is selective loss of the beta cells within the endocrine islets, as a consequence of T‐cell and cytokine mediated destruction of these cells, perhaps in conjunction with destruction of the peri‐islet Schwann cells. In type 2 diabetes, the etiology of the resistance ranges from post‐receptor defects in the insulin signaling pathway to excessive production of adipocyte derived cytokines that antagonize insulin action to mitochondrial defects that interfere with glucose disposal. Proteome based technologies are providing new insights into these defects.
Non Classical HLA Genes and Non‐HLA Genes in a Population of Infants at Familial Risk of Atopy
Authors: De Silvestri A. , Belloni C. , De Amici M. , Mazzola P. , Zorzetto M. , Martinetti M. , Salvaneschi L. , Cuccia M.
Published: June 2013

Aim: We investigated on parental history and IgE serum level in 2588 consecutive newborns to individuate babies “at risk” of atopy at birth and we analysed the polymorphisms of class III region to evaluate the association with immunogenetic markers of HLA: C4A, C4B, LTA, RAGE and TNFA genes; we performed TNF and IgE receptor (FCERB1) physiologically related gene polymorphisms.

Result: 791 babies/2588 (30.6%) were considered “at risk” for atopy and followed‐up: 400 had familial history of atopy (at least one parent or sibling), 256 had IgE > 0.35 kUA/l at birth and during the follow‐up and 135 were positive for both conditions.

The allele C4B2 was significantly more frequent in the sample of babies at risk (22.1% vs 10%, p < 0.001). Furthermore, the mean value of IgE at birth in babies carrying the allele C4B2 was 2.26 KUA/l versus 0.74 KUA/l in those not carrying this allele (p = 0.01). No signiﬁcant association emerged for RAGE at the centromeric end of class III region and for LTA, TNFA at the telomeric one. TNFRI, TNFRII and FCERB1 gene polymorphisms also seemed not implicated.

Conclusion: Our study confirms that HLA class III region seems involved in familial predisposition to atopy, and C4B gene probably acts as a marker of a more restricted subregion.
Relationship of the 1793G‐A and 677C‐T Polymorphisms of the 5,10‐Methylenetetrahydrofolate Reductase Gene to Coronary Artery Disease
Authors: Kebert Cory B. , Eichner June E. , Moore William E. , Schechter Eliot , Yaoi Takuro , Vogel Steve , Allen Richard A. , Dunn S. Terence
Published: June 2013

Numerous studies have investigated the relationship between polymorphisms, in particular 677C‐T and 1298A‐C, of the methylene‐tetrahydrofolate reductase (MTHFR) gene and coronary artery disease (CAD) with conflicting results. This study investigates the potential association of two point mutations in MTHFR, 677C‐T and 1793G‐A, along with other risk factors, with CAD. This is the first hospital‐based study to investigate 1793G‐A in this context. Genotype analysis was performed on 729 Caucasians and 66 African Americans undergoing coronary angiography using a novel PCR‐based assay involving formation of Holliday junctions. Allelic frequencies for 677C‐T were 66.2% C and 33.8% T for Caucasians and 90.9% C and 9.1% T for African Americans. With respect to the 1793G‐A polymorphism, allelic frequencies were 94.7% G and 5.3% A for Caucasians and 99.2% G and 0.8% A for African Americans. Disease associations were examined in the Caucasian patients due to their greater genotype variability and larger number in the patient cohort. Results suggest that neither 677CT heterozygotes (OR‐1.36; 95% CI 0.95 to 1.96) nor mutant homozygotes (OR‐0.73; 95% CI 0.44 to 1.20) have either an increased or decreased risk for CAD compared to the 677CC genotype. Likewise, the 1793GA genotype did not demonstrate a statistically significant association with CAD compared to 1793GG patients (OR‐0.79; 95% CI 0.47 to 1.33). Mean homocysteine levels (μmol/L) increased from normal to mutant for 677C‐T (677CC: 10.2; 677CT: 11.0; 677TT: 11.6) and normal to heterozygous in 1793G‐A (1793GG: 10.7; 1793GA: 11.5). These MTHFR polymorphisms did not contribute to the prediction of clinically defined CAD in Caucasians.
Higher Expression of Epidermal Growth Factor Receptor Is Associated with Extracellular Matrix Metalloprotease Inducer in Colorectal Adenocarcinoma: Tissue Microarray Analysis of Immunostaining Score with Clinicopathological Parameters
Authors: Jin Jong-Shiaw , Wu Chi-Ying , Lin Yeh-Feng , Wang Jia-Yi , Yu Cheng-Ping , Sheu Lai-Fa , Chiang Hung , Tsai Wen-Chiuan , Lee Wei-Hwa
Published: June 2013

Aim: Extracellular matrix metalloprotease inducer (EMMPRIN) expression was demonstrated in several cancers, but its expression profile in colorectal cancers remains unclear. Epidermal growth factor receptor (EGFR) was reported to regulate EMMPRIN expression in human epithelial cancers. Our purpose was to determine EMMPRIN expression and its relationship with EGFR in colorectal cancers.

Methods: Immunohistochemical analysis of EMMPRIN and EGFR was performed in tissue microarray slides of 90 surgical specimens including 32 well differentiated, 35 moderately differentiated, and 23 poorly differentiated colorectal adenocarcinomas.

Results: All colorectal adenocarcinomas showed significant immunohistochemical expression of EMMPRIN. The EMMPRIN scores in poorly differentiated (303 ± 21) and moderately differentiated (326 ± 17) colorectal adenocarcinoma were significantly higher than in well differentiated (166 ± 20) colorectal adenocarcinoma. EGFR expression was mainly on the cell surface of tumor cells and the immunostaining scores of EGFR were significantly associated with the advanced clinical T and N stages. A significantly positive relationship between EMMPRIN and EGFR immunostaining scores was also noted.

Conclusions: Increased expression of EMMPRIN and EGFR in colorectal adenocarcinomas is associated with clinicopathological parameters of advanced colorectal adenocarcinoma stages. In addition, the data from this study support the notion that EGFR expression may up-regulate EMMPRIN expression.
The Catalase –262C/T Promoter Polymorphism and Diabetic Complications in Caucasians with Type 2 Diabetes
Authors: dos Santos Kátia Gonçalves , Canani Luís Henrique , Gross Jorge Luiz , Tschiedel Balduíno , Souto Kátia Elisabete Pires , Roisenberg Israel
Published: June 2013

Catalase is a central antioxidant enzyme constituting the primary defense against oxidative stress. In this study, we investigated whether the functional –262C/T polymorphism in the promoter of catalase gene is associated with the presence of diabetic retinopathy (DR), diabetic nephropathy (DN) and ischemic heart disease (IHD) in 520 Caucasian‐Brazilians with type 2 diabetes. The –262C/T polymorphism was also examined in 100 Caucasian blood donors. Patients underwent a clinical and laboratory evaluation consisting of a questionnaire, physical examination, assessment of diabetic complications and laboratory tests. Genotype analysis was performed using the polymerase chain reaction followed by digestion with restriction enzyme. The genotype and allele frequencies of the –262C/T polymorphism in patients with type 2 diabetes were very similar to those of blood donors (T allele frequency = 0.20 and 0.18, respectively). Likewise, there were no differences in either genotype or allele frequencies between type 2 diabetic patients with or without DR, DN or IHD. Thus, our results do not support the hypothesis that the –262C/T polymorphism is related to the development of DR, DN or IHD in patients with type 2 diabetes. Further studies are necessary to elucidate the role of catalase gene polymorphisms in the pathogenesis of diabetic complications.
International Standard Reagents for HPV Detection
Authors: Pagliusi Sonia R. , Garland Suzanne M.
Published: June 2013

Humam papillomavirus is the commonest genital viral infection in healthy sexually active subjects, and the presence of chronic or persistent HPV types in genital cells may constitute a prognostic marker of underlying, or predict future HPV‐associated diseases. A variety of novel tests for detecting the presence of oncogenic HPV types in biological specimens have been reported. These are based on the various stages of infection and viral life cycle. HPV infects squamous epithelium with expression of various gene products intimately linked to epithelial cell differentiation. Hence, there are basically three classes of detectable markers directly derived from HPVs: molecular markers based on detection of nucleic acid sequences, serological markers based on detection of antibodies against viral proteins, and cellular markers based on detection of proteins expressed intracellularly, upon either infection or carcinogenesis. The nature of various assays and the development of international standard reagents for qualitative and quantitative assessment of assay performance are outlined. There is an increasing demand to develop standard tools to assess the quality of HPV detection systems, for regulatory and clinical management purposes. International standard reagents for HPV will help defining the analytical sensitivity and specificity of various detection methods, and will allow assuring that laboratory services used to evaluate disease burden, HPV vaccines, and cancer prevention strategies are accurate and comparable worldwide. The advancement of prophylactic vaccine candidates against HPV infections and related diseases stresses the increasing importance of HPV assays in monitoring the impact of HPV vaccination on disease burden.
Carbohydrate Markers in Colon Carcinoma
Authors: Szajda Sławomir Dariusz , Jankowska Anna , Zwierz Krzysztof
Published: June 2013

Spontaneously mutated multiple oncogenes and/or tumor suppressor genes in colon epithelial cell and its progeny, may cause proliferation out of control and create benign colon neoplasm (colon polyp). If additional mutations involve genes responsible for cell adhesion and movement, aberrant epithelial cells may become malignant (colon cancer) and invade surrounding and remote tissues, creating secondary tumors called metastases.

Incidence of colorectal cancer dramatically increases at 50–65 year of age. In Europe in 2006 colorectal cancer consisted 12.9% of all cancers and caused 207 400 deaths.

To laboratory detection and monitoring of colon cancer are used tumor markers. Tumor markers are substances produced by the body in response to cancer, or by cancer tissue itself. Glycoconjugate markers for colon cancer include aberrant: mucins covering the surface of the colon epithelial cells, cadherins, selectins and Ig –like adhesion molecules mediating cell‐cell adhesion, integrins and integral membrane proteoglycans responsible for adhesion of colon epithelial cells to extracellular matrix, glycoconjugate components of ECM, as well as lysosomal membrane glycoproteins and exoglycosidases.

Detection of colon cancer at early non malignant stage is crucial in its prevention and eradication. As colon cancer is the effect of accumulation many somatic mutations in oncogens, supressors, mismatch repair genes and many genes responsible for posttranslational modifications of proteins, multidirectional approach should be applied for its detection. A glycobiological approach to diagnosis and treatment of colorectal cancer should be directed to detection changes in glycosylation accompanying every step of colon cancer progression, and correlation between changes in glycosylation and tumor progression.
TGF‐β Signaling in Neuronal Stem Cells
Authors: Yun Chohee , Mendelson Jonathan , Blake Tiffany , Mishra Lopa , Mishra Bibhuti
Published: June 2013

Transforming growth factor beta (TGF‐β) signaling has diverse and complex roles in various biological phenomena such as cell growth, differentiation, embryogenesis and morphogenesis. ES cells provide an essential model for understanding the role of TGF‐β signaling in lineage specification and differentiation. Recent studies have suggested significant role of TGF‐β in stem/progenitor cell biology. Here in this review, we focus on the role of the TGF‐β superfamily in neuronal development.
GPR87 Is an Overexpressed G‐Protein Coupled Receptor in Squamous Cell Carcinoma of the Lung
Authors: Gugger Mathias , White Richard , Song Susan , Waser Bea , Cescato Renzo , Rivière Pierre , Reubi Jean Claude
Published: June 2013

Lung cancer is the leading cause of cancer death worldwide. The overall 5‐year survival after therapy is about 16% and there is a clear need for better treatment options, such as therapies targeting specific molecular structures. G‐protein coupled receptors (GPCRs), as the largest family of cell surface receptors, represent an important group of potential targets for diagnostics and therapy. We therefore used laser capture microdissection and GPCR‐focused Affymetrix microarrays to examine the expression of 929 GPCR transcripts in tissue samples of 10 patients with squamous cell carcinoma and 7 with adenocarcinoma in order to identify novel targets in non‐small cell lung carcinoma (NSCLC). The relative gene expression levels were calculated in tumour samples compared to samples of the neighbouring alveolar tissue in every patient. Based on this unique study design, we identified 5 significantly overexpressed GPCRs in squamous cell carcinoma, in the following decreasing order of expression: GPR87 > CMKOR1 > FZD10 > LGR4 > P2RY11. All are non‐olfactory and GRAFS (glutamate, rhodopsin, adhesion, frizzled/taste2, secretin family) classified. GPR87, LGR4 and CMKOR1 are orphan receptors. GPR87 stands out as a candidate for further target validation due to its marked overexpression and correlation on a mutation‐based level to squamous cell carcinoma.
An Increase in N‐Ras Expression is Associated with Development of Hormone Refractory Prostate Cancer in a Subset of Patients
Authors: Traynor Pamela , McGlynn Liane M. , Mukhergee Rono , Grimsley Samuel J. S. , Bartlett John M. S. , Edwards Joanne
Published: June 2013

Protein expression of H, K and N‐Ras was assessed in hormone sensitive and hormone refractory prostate tumour pairs from 61 patients by immunohistochemistry. Expression of H‐Ras and K‐ Ras was not associated with any known clinical parameters. In contrast an increase in N‐Ras membrane expression in the transition from hormone sensitive to hormone refractory prostate cancer was associated with shorter time to relapse (p = 0.01) and shorter disease specific survival (p = 0.008). In addition, patients with an increase in N‐Ras membrane expression had lower levels of PSA at relapse (p = 0.02) and expression correlated with phosphorylated MAP kinase (p = 0.010) and proliferation index (Ki67, p = 0.02). These results suggest that in a subgroup patients N‐Ras expression is associated with development of hormone refractory prostate cancer via activation of the MAP kinase cascade.
Elevation in Peripheral Blood Circulating Tumor Cell Number Correlates with Macroscopic Progression in UICC Stage IV Colorectal Cancer Patients
Authors: Molnar Bela , Floro Lajos , Sipos Ferenc , Toth Bernadett , Sreter Lydia , Tulassay Zsolt
Published: June 2013

Aims: Cytokeratin(CK) based real‐time RT‐PCR assays (QRT‐PCR) are now available for peripheral blood circulating tumor cell (CTC) evaluations in colorectal cancer(CRC) patients. Results are non‐existent for the application of these techniques to the determination of progression and therapy response in Dukes stage D CRC patients.

Patients and methods: Each month 30 ml peripheral blood of 30 Dukes D patients (17 with progression) were drawn. CEA, CA19‐9, CA72‐A and TPA‐M determinations were made. CK20, thymidilate synthase(TS) QRT‐PCR was performed, as well. Buffy coat was used for immunmagnetic cancer cell isolation and CTC counting. Correlation between elevated CTC and macroscopic progression within 3 months was determined by Chi2 test.

Results: Microscopic CTC single cell, doublet, cluster number were found in correlation with CK20 QRT‐PCR results (p <
0.01). There was a significant increase in microscopic CTC number, CK20 and decrease in TS QRT‐PCR levels (p < 0.05) in
the peripheral blood of the non‐responder as compared to responder patients. Elevation of the CTC was in significant correlation
with macroscopic progression of the disease in 3 months (p < 0.01).

Conclusions: CTC number reflects the chemotherapeutic sensitivity of CRC patients. Elevation of circulating tumor cell number in peripheral blood is in correlation with macroscopic progression.
Multimarker Approach in Cardiovascular Risk Prediction
Authors: Ikonomidis Ignatios , Michalakeas Christos A. , Lekakis John , Paraskevaidis Ioannis , Th. Kremastinos Dimitrios
Published: June 2013

Various biomarkers express different pathways and pathophysiologic mechanisms of cardiovascular disease, such as inflammation, oxidative stress, myocardial injury, activation of the neurohormonal pathways, myocardial stress and renal function. Current thinking supports the notion that the combination of these biomarkers could increase their diagnostic and prognostic value. The multimarker approach offers benefits since it increases the diagnostic and prognostic information and may help in the design of a strategy for prevention or management of cardiovascular diseases. The purpose of the current review is to describe the characteristics of promising biomarkers which have shown an important additive value in the assessment of cardiovascular risk. Also, an extended reference is made regarding studies that address the prognostic value of multimarker models in the settings of primary prevention of cardiovascular disease and secondary prevention for patients with acute coronary syndromes, chronic coronary artery disease and heart failure.
Investigation of CBS, MTR, RFC-1 and TC Polymorphisms as Maternal Risk Factors for Down Syndrome
Authors: Fintelman-Rodrigues N. , Corrêa J. C. , Santos J. M. , Pimentel M. M. G. , Santos-Rebouças C. B.
Published: June 2013

Recent evidence shows that almost 92% of the DS children are born from young mothers, suggesting that other risk factors than advanced maternal age must be involved. In this context, some studies demonstrated a possible link between DS and maternal polymorphisms in genes involved in folate metabolism. These polymorphisms, as well as low intake of folate could generate genomic instability, DNA hypomethylation and abnormal segregation, leading to trisomy 21. We compared the frequency ofCBS844ins68,MTR2756A>G,RFC-1 80G> A andTC776C>G polymorphisms among 114 case mothers and 110 matched controls, in order to observe whether these variants act as risk factors for DS. The genotype distributions revealed that there were not significant differences between both samples. However, when we proceed the multiplicative interaction analyses between the four polymorphisms described above together with the previously studiedMTHFR677C>T,MTHFR1298A>C andMTRR66A>G polymorphisms, our results show that the combined genotypeTC776CC /MTHFR677TT andTC776CC /MTR2756AG were significantly higher in the control sample. Nevertheless, there was no significant association after Bonferroni correction. Our results suggest that maternal folate-related polymorphisms studied here have no influence on trisomy 21 susceptibility in subjects of Brazilian population.
A novel Multiple-Marker Method for the Early Diagnosis of Oral Squamous Cell Carcinoma
Authors: Ries Jutta , Mollaoglu Nur , Toyoshima Takeshi , Vairaktaris Eleftherios , Neukam Friedrich W. , Ponader Sabine , Nkenke Emeka
Published: June 2013

Objective: Melanoma associated antigens-A (MAGE-A) expression is highly specific to cancer cells. Thus, they can be the most suitable targets for the diagnosis of malignancy. The aim of this study was to evaluate the sensitivity of multiple MAGE-A expression analysis for the diagnosis of oral squamous cell carcinoma (OSCC).

Methods: Total of 70 OSSC and 20 normal oral mucosal (NOM) samples of otherwise healthy volunteers were examined for the expression of 10 different single antigens out of 12 different MAGE-A subtypes by highly sensitive reverse transcriptase polymerase chain reaction (RT-PCR) methods. The results were correlated to clinicopathological parameters of tumor samples.

Results: Expression of MAGE-A was restricted to OSCC. The expression frequency of single antigen was between 10% and 55%. However, expression rate was increased up to 93% by the elevated number of genes examined. A significant correlation was found between the expression of MAGE-A and malignancy (p = 0.0001). In addition, multiple MAGE-A detection has also correlated to the incidence of lymph node metastasis, grading and advanced clinical stages.

Conclusions: Analysis of multiple MAGE-A expression is more sensitive than the analysis of a single MAGE-A for the diagnostic evaluation of OSCC. Multiple MAGE-A expression analysis may be a very sensitive method to be used for the diagnosis even in the early stage of OSCC.
GSTM1,GSTM3andGSTT1Gene Variants and Risk of Benign Prostate Hyperplasia in North India
Authors: Mittal Rama Devi , Kesarwani Pravin , Singh Ranjana , Ahirwar Dinesh , Mandhani Anil
Published: June 2013

Glutathione S-transferases (GSTs) play an important role in detoxification of various toxic compounds like carcinogens in cigarette smoke and tobacco by conjugating to toxic compounds and inactivating their hazardous effect. Variation in Glutathione S-Transferases (GSTs) genes may alter the catalytic efficiency of GST isoenzymes leading to potential increase in cancer susceptibility due to various carcinogens. We therefore, investigated association of GSTM1, GSTM3 and GSTT1 variants with susceptibility to benign prostate hyperplasia (BPH) and cigarette, tobacco chewing and alcohol consumption as confounding factors in 141 BPH and 184 healthy controls. Results showed increased risk for BPH susceptibility in patients with GSTM1 null genotype (OR-2.03,p= 0.013) and smoking (OR-3.12,p= 0.028), tobacco chewing (OR-2.54,p= 0.039) and alcohol habits (OR-3.39,p= 0.010). Null genotype of GSTM1 with cigarette, tobacco and alcohol habits predisposed increased risk for BPH.
A Homogeneous HLA-B*27 Genotyping Assay Using Dried Reagent Mixtures
Authors: Kiviniemi Minna , Ilonen Jorma , Lövgren Timo
Published: June 2013

The presence of HLA-B*27 allele with patients suspected with ankylosing spondylitis can be used in the diagnostic process. We have developed an assay for typing for the HLA-B*27 in whole blood dried on sample collection cards using pre-dried reagent wells and homogeneous time-resolved fluorescence based PCR approach. Essentially only the sample needs to be added to the dry ready-to-use reaction well in order to start the homogenous amplification assay. The method was validated with 229 samples also typed with an existing DELFIA-based method and results of both assays were 100% concordant. The dried reagents were shown to be stable at least up to eight weeks at room temperature without any decline in their performance.
Helicobacter pylori: Determinant and Markers of Virulence
Authors: Mascellino M. T. , Margani M. , Oliva A.
Published: June 2013

In this review, we examine the main virulence determinants inHelicobacter pylori(Hp) strains and the correlation between these and the different diseases followingHpinfections. We also discuss the host response toHpand the implications and advantages of the development of non-invasive pre-endoscopy screening of molecular and serological markers associated withHpvirulence and disease progression putting an emphasis on new screening techniques such as the Luminex – X multi analytes profiling technology and the multiplex PCR.