bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunohistochemical Expression of Retinoblastoma Gene Product in Normal, Hyperplastic and Malignant Endometrium. Correlation with p53 Protein Expression, c‐erbB‐2, Hormone Receptors’ Status and Proliferative Activity
Authors: Ioachim E. E. , Goussia A. C. , Kitsiou E. G. , Charalabopoulos K. , Mermiga E. , Stefanaki S.
Published: June 2013

Alterations of the retinoblastoma (Rb) gene have been described in several human neoplasms and recently, it has been suggested that these alterations may play a role in the development of endometrial carcinomas. Paraffin sections from 31 cases of normal endometrium (16 proliferative, 15 secretory), 35 hyperplastic lesions and 89 endometrial carcinomas were investigated immunohistochemically for Rb protein (pRb) expression. The results were compared with p53 and c‐erbB‐2 protein expression, estrogen (ER) and progesterone (PR) receptors’ status and with clinicopathological prognostic factors. pRb was expressed in normal, hyperplastic and neoplastic epithelium. Proliferative endometrium showed more intense and extensive pRb staining than secretory endometrium. pRb reactivity was heterogeneous in the hyperplastic endometrial cells. Lack or focal (< 10% of endometrial cells) pRb immunostaining was noted in 56.2% and 27% of carcinomas, respectively. In the remaining cases (16.8%) pRb staining was heterogeneous or diffuse. The absence or presence of pRb expression was independent of grade and stage. In normal proliferative and secretory endometrium, pRb expression was correlated with PR (p = 0.006 and p = 0.001, respectively), PCNA (p = 0.04 and p = 0.01, respectively) and MIB1 (p = 0.02 and p < 0.0001, respectively) expression. In hyperplasias, pRb was related to PR (p = 0.016) and MIB1 (p < 0.0001) expression. In carcinomas, a relationship of pRb expression with p53 (p = 0.0015), ER (p = 0.0002), PR (p = 0.0004) and PCNA (p = 0.013) status was detected. We suggest that the absence or presence of pRb expression does not seem to be associated with the progression of endometrioid carcinoma. In addition, pRb seems to be normally regulated in relation to the proliferative growth fraction of the tumours.
Shotgun Proteomics and Biomarker Discovery
Authors: McDonald W. Hayes , Yates John R.
Published: June 2013

Coupling large‐scale sequencing projects with the amino acid sequence information that can be gleaned from tandem mass spectrometry (MS/MS) has made it much easier to analyze complex mixtures of proteins. The limits of this “shotgun” approach, in which the protein mixture is proteolytically digested before separation, can be further expanded by separating the resulting mixture of peptides prior to MS/MS analysis. Both single dimensional high pressure liquid chromatography (LC) and multidimensional LC (LC/LC) can be directly interfaced with the mass spectrometer to allow for automated collection of tremendous quantities of data. While there is no single technique that addresses all proteomic challenges, the shotgun approaches, especially LC/LC‐MS/MS‐based techniques such as MudPIT (multidimensional protein identification technology), show advantages over gel‐based techniques in speed, sensitivity, scope of analysis, and dynamic range. Advances in the ability to quantitate differences between samples and to detect for an array of post‐translational modifications allow for the discovery of classes of protein biomarkers that were previously unassailable.
Evaluation of Nitric Oxide (NO) Levels in Hepatitis C Virus (HCV) Infection: Relationship to Schistosomiasis and Liver Cirrhosis among Egyptian Patients
Authors: Hassan Mahmoud Ismail , Kassim Samar Kamal , Ali Hebatalla Said , Sayed El-Dieb Abd ElSattar , Khalifa Ali
Published: June 2013

Nitric oxide (NO), a recently discovered free radical, is overproduced in liver cirrhosis. Hepatitis C virus (HCV) might increase NO levels via increased inducible NO synthase (iNOS). This work was carried out to study the effect of HCV‐induced liver cirrhosis on NO levels among Egyptian patients. The study included 46 patients with liver cirrhosis, and 30 healthy individuals of matched age and sex. NO levels determined as the stable endproduct nitrate, showed a statistically significant increase among patients compared to the control group (P < 0.001). Furthermore, NO levels increased proportionally with the severity of liver cirrhosis as assessed by Child’s classification (P < 0.05). Moreover, schistosomial infection enhanced NO levels in cirrhotic patients with HCV infection compared to non‐bilharzial patients (P < 0.001). Polymerase chain reaction (PCR) and branched DNA assays were used for detection of HCV RNA positivity, and measurement of the virus load, respectively. Both showed a positive correlation with the NO levels (P < 0.001). At a nitrate cutoff value of 70 μmol/L, the sensitivity and specificity were 83.0% and 37.0% respectively. Chi square analysis showed a significant correlation between ALT levels and both HCV RNA positivity by polymerase chain reaction (PCR) (P < 0.02), and virus load (P < 0.05). Interestingly enough, there was a significant positive correlation between HCV RNA and schistosomal antibody titer as measured by hemaglutination inhibition assay (HAI) (P < 0.05). The data presented in this report indicated an association between NO levels and the development and progression of liver cirrhosis. Furthermore, the findings obtained from this study demonstrated that schistomiasis is an important risk factor involved in enhancement of NO levels and virus replication. The latter may aggravate liver cell injury and hence the development of cirrhosis.
Animal Models for imaging
Authors: Croft Barbara Y.
Published: June 2013

Animal models can be used in the study of disease. This chapter discusses imaging animal models to elucidate the process of human disease. The mouse is used as the primary model. Though this choice simplifies many research choices, it necessitates compromises for in vivo imaging. In the future, we can expect improvements in both animal models and imaging techniques.
Bioassays Based on Molecular Nanomechanics
Authors: Majumdar Arun
Published: June 2013

Recent experiments have shown that when specific biomolecular interactions are confined to one surface of a microcantilever beam, changes in intermolecular nanomechanical forces provide sufficient differential torque to bend the cantilever beam. This has been used to detect single base pair mismatches during DNA hybridization, as well as prostate specific antigen (PSA) at concentrations and conditions that are clinically relevant for prostate cancer diagnosis. Since cantilever motion originates from free energy change induced by specific biomolecular binding, this technique is now offering a common platform for label‐free quantitative analysis of protein‐protein binding, DNA hybridization DNA‐protein interactions, and in general receptor‐ligand interactions. Current work is focused on developing “universal microarrays” of microcantilever beams for high‐throughput multiplexed bioassays.
Peroxisomal Enzymes and 8‐Hydroxydeoxyguanosine in Rat Liver Treated with Perfluorooctanoic Acid
Authors: Abdellatif Awad , Al-tonsy Ahmad H. , Awad Mahmoud E. , Roberfroid M. , Khan M. Nezam Ullah
Published: June 2013

Although peroxisome proliferators are considered non‐genotoxic agents, most of them, nevertheless, were found to promote and/or induce, hepatocellular carcinoma (HCC) in rodents. The aim of the present study is, first, to investigate whether the peroxisome proliferator perfluorooctanoic acid (PFOA) possesses inherent liver cancer promoting activity, and second, to study the possible mechanisms involved. To acheive these aims two protocols have been applied, a biphasic protocol (initiation by diethyl‐nitrozamine (DEN) 200 mg/kg i.p. followed by treatment with 0.005% or 0.02% perflourooctanoic acid (PFOA) for 14 and 25 weeks) and a triphasic initiation, selection‐promotion (IS) protocol (initiation by giving 200 mg/kg DEN i.p. followed by a selection procedure for 2 weeks consisting of giving 0.03% 2‐acetylaminofluorene (2‐AAF) in diet). In the middle of this treatment a single oral dose of carbon tetrachloride (2.0 ml/kg) was given, followed by giving diet containg 0.015% of PFOA for 25 weeks. After applying both protocols, our results showed slight increase in the catalase activity while acyl CoA oxidase activity was markedly increased. Both experiments indicated that PFOA has a liver cancer promoting activity. Other groups of rats were given either basal diet or diet containing 0.02% PFOA. Five or nine weeks later they were sacrificed and the levels of 8‐hydroxydeoxyguanosine in the isolated DNA were estimated. The data showed a slight nonetheless insignificant increase in 8‐hydroxydeoxyguanosine. From the present data, it is concluded that PFOA is a true liver cancer promoter that may not require extensive initial DNA damage for its promoting activity.
Haplotype Analysis of Norwegian and Swedish Patients with Acute Intermittent Porphyria (AIP): Extreme Haplotype Heterogeneity for the Mutation R116W
Authors: Tjensvoll Kjersti , Bruland Ove , Floderus Ylva , Skadberg Øyvind , Sandberg Sverre , Apold Jaran
Published: June 2013

Acute intermittent porphyria (AIP), the most common of the acute porphyrias, is caused by mutations in the gene encoding hydroxymethylbilane synthase (HMBS) also called porphobilinogen deaminase (PBGD). The mutation spectrum in the HMBS gene is characterized by a majority of family specific mutations. Among the exceptions are R116W and W198X, with high prevalence in both the Dutch and Swedish populations. These two mutations were also detected in unrelated Norwegian patients. Thus, Norwegian and Swedish patients were haplotyped using closely linked flanking microsatellites and intragenic single nucleotide polymorphisms (SNPs) to see if the high frequency of these two mutations is due to a founder effect. Twelve intragenic SNPs were determined by a method based on fluorescent restriction enzyme fingerprinting single‐strand conformation polymorphism (F‐REF‐SSCP).

W198X occurred exclusively on one haplotype in both Norwegian and Swedish patients, showing that it has originated from a common gene source. In contrast, R116W was found on three different haplotypes in three Norwegian families, and in five Swedish families on four or five haplotypes. This extreme haplotype heterogeneity indicates that R116W is a recurrent mutation, maybe explained by the high mutability of CpG dinucleotides. This can also explain why it is the only AIP mutation reported to occur in seven different populations (Norway, Sweden, Finland, Netherlands, France, Spain and South Africa).
Role of the Pathologist in the Diagnosis of Hereditary Non‐Polyposis Colorectal Cancer
Authors: Jass Jeremy R.
Published: June 2013

The aim of this paper is to indicate how the pathologist may suspect a diagnosis of hereditary non‐polyposis colorectal cancer (HNPCC) on the basis of histological criteria and patient age alone. A single morphological feature, namely the presence of intra‐epithelial lymphocytes (tumor infiltrating lymphocytes), identifies the majority of colorectal cancers (CRC) with the DNA microsatellite instability‐high phenotype. A number of pathological criteria can help to distinguish HNPCC from sporadic MSI‐H CRC, though age below 60 years is an important pointer towards HNPCC. Immunohistochemistry to demonstrate loss of expression of DNA mismatch repair genes serves as a highly reliable test of mismatch repair deficiency if antibodies to hMLH1, hMSH2, hMSH6 and hPMS2 are employed.
Diagnostic Application of hMLH1 Methylation in Hereditary Non‐Polyposis Colorectal Cancer
Authors: Matsubara Nagahide
Published: June 2013

Colorectal cancer (CRC) due to mismatch repair (MMR) defect has distinct characteristics among unselected CRCs. These CRCs are biologically less aggressive and, thus, showing better prognosis but less sensitive to the 5FU‐based chemotherapy. CRCs with MMR defect derive from both hereditary and sporadic reasons. Germline inactivation of MMR genes (hMLH1, hMSH2, hMSH6, and hPMS2) underlies the hereditary CRC with MMR defect (Lynch syndrome) and epigenetic silencing of hMLH1 gene causes the sporadic CRC with MMR defect. Hereditary and sporadic CRC with MMR defect can be detectable by microsatellite instability (MSI) test or immunohistochemical analysis among general CRCs. Lynch syndrome can be diagnosed by the clinical criteria or by genetic test to detect pathogenic germline mutations in MMR genes. However, both clinical criteria and genetic test are inadequate for the diagnosis of Lynch syndrome. Since genetic test for the diagnosis of the Lynch syndrome is expensive and not always identify pathogenic germline mutations, effective and inexpensive screening program is desirable. Here we propose a possible application of methylation test combined with MSI or pathological analysis as an effective and a cost‐saving new strategy for screening of Lynch syndrome.
The Role of Diffuse Optical Spectroscopy in the Clinical Management of Breast Cancer
Authors: Shah Natasha , Cerussi Albert E. , Jakubowski Dorota , Hsiang David , Butler John , Tromberg Bruce J.
Published: June 2013

Diffuse optical spectroscopy (DOS) of breast tissue provides quantitative, functional information based on optical absorption and scattering properties that cannot be obtained with other radiographic methods. DOS‐measured absorption spectra are used to determine the tissue concentrations of deoxyhemoglobin (Hb‐R), oxyhemoglobin (Hb‐O2), lipid, and water (H2O), as well as to provide an index of tissue hemoglobin oxygen saturation (StO2). Tissue‐scattering spectra provide insight into epithelial, collagen, and lipid contributions to breast density. Clinical studies of women with malignant tumors show that DOS is sensitive to processes such as increased tissue vascularization, hypoxia, and edema. In studies of healthy women, DOS detects variations in breast physiology associated with menopausal status, menstrual cycle changes, and hormone replacement. Current research involves using DOS to monitor tumor response to therapy and the co‐registration of DOS with magnetic resonance imaging. By correlating DOS‐derived parameters with lesion pathology and specific molecular markers, we anticipate that composite “tissue optical indices” can be developed that non‐invasively characterize both tumor and normal breast‐tissue function.
The Future of Medical Imaging in the Detection of Early Markers of Disease
Authors: Croft Barbara Y.
Published: June 2013

Imaging techniques are a combination of a contrast mechanism, exogenous or endogenous, and an instrument to exploit that contrast. This final chapter of these two special issues of this journal points to possible ways to improve the ability of imaging systems to exploit markers of cancer in the early detection of that disease. The aim not only is to find cancer at an earlier, more treatable stage, but to determine whether the disease discovered is dangerous and to indicate the possibilities for successful treatment. These topics are explored for each imaging system, with an emphasis on directions for future improvements.
Promise and Challenge: Markers of Prostate Cancer Detection, Diagnosis and Prognosis
Authors: Troyer D. A. , Mubiru J. , Leach R. J. , Naylor S. L.
Published: June 2013

Approximately 1 man in 6 will be diagnosed with prostate cancer during his life lifetime, and over 200,000 men in the U.S. are diagnosed with prostate cancer annually. Since the widespread adoption of PSA testing, about 60–70% of men at risk in the U.S. have had a blood test for prostate cancer. With this, prostate cancer death rates have decreased, yet only slightly. Thirty thousand men still die each year from this disease. PSA testing fails to identify a small but significant proportion of aggressive cancers, and only about 30% of men with a “positive” PSA have a positive biopsy. Additionally, of men who are treated for prostate cancer, about 25% require additional treatment, presumably due to disease recurrence. Also of concern is the growing evidence that there are some prostate cancers for which treatment may not be necessary. Very long‐term studies from the U.S. and Europe, following men with prostate cancer have found that some tumors do not progress over time. In these individuals, prostate cancer treatment is unnecessary and harmful as these men do not benefit from treatment but will be at risk of treatment‐related side effects and complications. They suggest a fundamental problem with prostate cancer: it is not possible, at this time, to predict the natural history of the disease. It is for these reasons that the most important challenge in prostate cancer today is the inability to predict the behavior of an individual tumor in an individual patient. Here we review issues related to performance and validation of biomarkers with a focus on “doing no harm”, and bearing in mind that it is the ultimate goal of early detection to save lives. Improved diagnostic and prognostic biomarkers are needed for prostate cancer, and the use of these markers should ultimately translate into increased life span and quality of life. The ultimate goal would be to not only have accurate biomarkers suitable for early diagnosis, but also biomarkers that identify men at greatest risk of developing aggressive disease. Technology has been brought to bear on this problem, and the major approaches are genomics, expression analysis, and proteomics. Proteomics and DNA methylation assays may soon be used in sensitive and specific diagnostic testing of serum and tissues for cancer. Expression arrays may be used to establish both a more specific diagnosis and prognosis for a particular tumor. The proteome is only beginning to be understood, and alternative splicing and post‐translational modifications of proteins such as glycosylation and phosphorylation are challenging areas of study. Finally, risk assessment and prognosis are being pursued through analysis of genomic polymorphisms (single nucleotide polymorphisms, SNPs). This huge task is only beginning, and requires the combined expertise of molecular epidemiologists, oncologists, surgeons, pathologists, and basic scientists.
Serum Protein Expression Profiling for Cancer Detection: Validation of a SELDI‐Based Approach for Prostate Cancer
Authors: Grizzle William E. , Adam Bao-Ling , Bigbee William L. , Conrads Thomas P. , Carroll Christopher , Feng Ziding , Izbicka Elzbieta , Jendoubi Moncef , Johnsey Donald , Kagan Jacob , Leach Robin J. , McCarthy Diane B. , Semmes O. John , Srivastava Shiv , Srivastava Sudhir , Thompson Ian M. , Thornquist Mark D. , Verma Mukesh , Zhang Zhen , Zou Zhiqiang
Published: June 2013

Multiple studies have reported that analysis of serum and other bodily fluids using surface enhanced laser desorption/ionization time of flight mass spectroscopy (SELDI‐TOF‐MS) can identify a “fingerprint” or “signature” of spectral peaks that can separate patients with a specific disease from normal control patients. Ultimately, classification by SELDI‐TOF‐MS relies on spectral differences in position and amplitude of resolved peaks. Since the reproducibility of quantitation, resolution and mass accuracy of the SELDI‐TOF‐MS, or any high throughput mass spectrometric technique, has never been determined this method has come under some skepticism as to its clinical usefulness. This manuscript describes a detailed design of a three‐phase study to validate the clinical usefulness of SELDI‐TOF‐MS in the identification of patients with prostatic adenocarcinoma (PCA). At the end of this validation study, the usefulness of the general SELDI‐TOF‐MS approach to identifying patients with PCA will be demonstrated and how it compares with PCA diagnosis by measuring prostate specific antigen.
Mutation Rate at Commonly Used Forensic STR Loci: Paternity Testing Experience
Authors: Aşıcıoğlua Faruk , Oguz-Savran Fatma , Ozbek Ugur
Published: June 2013

Paternity tests are carried out by the analysis of hypervariable short tandem repeat DNA loci. These microsatellite sequences mutate at a higher rate than that of bulk DNA. The occurrence of germline mutations at STR loci posses problems in interpretation of resulting genetic profiles. We recently analyzed 59–159 parent/child allele transfers at 13 microsatellite loci. We identified 12 mutations in 7 microsatellite loci. No mutations were occurred in other 6 loci. The highest mutation rate was observed with 5 mutations at D8S1179 locus at different alleles. The event was always single repeat related. The mutation rate was between 0 and 1.5 x 10-2 per locus per gamete per generation. The mutation event is very crucial for forensic DNA testing and accumulation of STR mutation data is extremely important for genetic profile interpretation.
Proteomics Approaches to Identify Tumor Antigen Directed Autoantibodies as Cancer Biomarkers
Authors: Imafuku Yuji , Omenn Gilbert S. , Hanash Samir
Published: June 2013

The identification of autoantibodies to tumor cell proteins by proteomics approaches has great potential impact on cancer biomarker discovery. The humoral immune response represents a form of biological amplification of signals that are otherwise weak due to very low concentrations of antigen, especially in the early stages of cancers. In addition, proteomics can detect immunoreactivity directed against protein post‐translational modifications. Two‐dimensional gel based Western blots, protein antigen microarrays, and multiplex ELISA reactions have been applied by our group to antigen based biomarker detection and validation. The latter two are based on liquid‐phase separations that are suitable for automation. This work has resulted in the identification of numerous cancer biomarker candidates. Large clinical studies are currently planned to establish their value in early cancer diagnosis.
Clinical Interest of the Combined Use of Serum CD26 and Alpha‐L‐Fucosidase in the Early Diagnosis of Colorectal Cancer
Authors: Ayude Daniel , de la Cadena María Páez , Cordero Oscar Javier , Nogueira Montserrat , Ayude José , Fernández-Briera Almudena , Rodríguez-Berrocal Francisco Javier
Published: June 2013

The purpose of this study was to assess if the combination of CD26 and alpha‐L‐fucosidase has a role in the diagnosis of colorectal cancer, paying particular attention to the stages in which the tumour is not yet disseminated. CD26 concentration and alpha‐L‐fucosidase activity were determined in sera from 110 colorectal cancer patients and 46 donors. The combination of CD26 and alpha‐L‐fucosidase showed a specificity of 100% with a sensitivity of 64% in the diagnosis of colorectal cancer. Interestingly, the combination of both markers had a sensitivity of 75% in the stage I at the highest specificity (100%), providing also high sensitivity levels for the other non‐disseminated stages (66% for stages II and III). In conclusion, the combined use of CD26 and alpha‐L‐fucosidase offers high sensitivity with high specificity in the diagnosis of colorectal cancer, especially at the earliest stage (TNM I).
Cervical Acid Phosphatase: A Biomarker of Cervical Dysplasia and a Potential Surrogate Endpoint for Colposcopy
Authors: Markovic Olivera , Markovic Nenad
Published: June 2013

Background: In 2000, cervical acid phosphatase (CAP) has been recently described as a biomarker labeling abnormal squamous cells on Pap smears (USPTO #6,143,512). The enzyme activity is presented as a red, granular deposit on a modified Papanicolaou background. This unique property was utilized for development of MarkPap® technology intended for cervical cancer screening.

Material/patients & methods: We conduct a multicenter, random assignment, assessor blinded, 2‐group (test and control), split‐sample designed clinical trial on 1,500 subject/specimens to assess safety and efficacy of the new test, in comparison with the control, for cervical cancer screening in standard Pap test environment. Safety is measured with frequency, severity and relation of adverse events. Efficacy is measured with primary endpoints (portion of positive/abnormal specimens detected, and the false negative rate). At the end of the follow‐up period (two years) when the study will be completed, other efficacy endpoints such as accuracy (sensitivity/specificity) and predictive values will be added to the method evaluation. Here we present in interim analysis.

Results: In April 2003, the recruitment was completed and the first twelve hundred cases have been evaluated. There was no serious or related adverse event in both groups. Minor, unrelated adverse events were rare and insignificantly distributed in both groups.

Primary endpoints: A: Portion of positive/abnormal specimens detected: Pe (new test) = 0.166, Ps (Pap control): 0.082; Ps’ (ACS reported value for US in year 2000): 0.07. Pe ≥ Ps + δ, for δ; = 0.5Ps. B. False negative rate: Pe = 0.05, Ps’ = 0.10. Confidence intervals: 95% CI: Test [0.148–0.193], Pap control [0.068–0.098]. OR = 2.26. χ2 = 40.69101 is greater than the critical value of 3.841 (P < 0.01).

Conclusion: We concluded that CAP had added to visibility of Pap test and has enabled cytoscreeners to significantly improve the detection of positive/abnormal specimens and reduce false negative rate. We discuss this unique property of CAP with emphasis on using it as a surrogate endpoint for colposcopy and eventual removal of a cervical lesion that, if untreated, could progress into cancer.
Receiver Operating Characteristic (ROC) to Determine Cut‐Off Points of Biomarkers in Lung Cancer Patients
Authors: Weiss Heidi L. , Niwas Santosh , Grizzle William E. , Piyathilake Chandrika
Published: June 2013

The role of biomarkers in disease prognosis continues to be an important investigation in many cancer studies. In order for these biomarkers to have practical application in clinical decision making regarding patient treatment and follow‐up, it is common to dichotomize patients into those with low vs. high expression levels. In this study, receiver operating characteristic (ROC) curves, area under the curve (AUC) of the ROC, sensitivity, specificity, as well as likelihood ratios were calculated to determine levels of growth factor biomarkers that best differentiate lung cancer cases versus control subjects. Selected cut‐off points for p185erbB-2 and EGFR membrane appear to have good discriminating power to differentiate control tissues versus uninvolved tissues from patients with lung cancer (AUC = 89% and 90%, respectively); while AUC increased to at least 90% for selected cut‐off points for p185erbB-2 membrane, EGFR membrane, and FASE when comparing between control versus carcinoma tissues from lung cancer cases. Using data from control subjects compared to patients with lung cancer, we presented a simple and intuitive approach to determine dichotomized levels of biomarkers and validated the value of these biomarkers as surrogate endpoints for cancer outcome.
Utility of Free/Total Prostate Specific Antigen (f/t PSA) Ratio in Diagnosis of Prostate Carcinoma
Authors: Thakur V. , Singh P. P. , Talwar M. , Mukherjee U.
Published: June 2013

The discovery that PSA exists in serum in both free and complexed forms led to development of immunoassays specific for different PSA forms. This helped in measuring free PSA in the presence of PSA‐ACT (PSA‐α antichymotrypsin), hence it was possible to calculate the percent free PSA or free to total PSA ratio, measurement of which was helpful in reducing the number of unnecessary biopsies significantly, while maintaining a high clinical sensitivity for detection of cancer. The study was performed on 103 consecutive male patients (mean age 68 ± 10.8 years SD) comprising of 90 patients with benign disease (87%) and 13 prostate carcinoma patients (13%), who had histologically proven prostate cancer. Patients with total PSA between 2–25 ng/ml were included in the study. 30 normal healthy males with age 58 ± 10 years, served as control. Serum total PSA and free PSA were analyzed using streptavidin biotin EIA method (M/s Roche Diagnostics, Germany). The mean total PSA in normal healthy control subjects was 1.86 ± 1.07 ng/ml. It was increased significantly in diseased condition. Its mean concentration in carcinoma patients was 12.6 ± 5.3 ng/ml and in benign patients it was 6.3 ± 4.6 ng/ml. The free to total PSA ratio in all the three groups was significantly different (p Combination of this ratio cutoff with other parameters like serum total PSA, DRE and TRUS helped in increasing the sensitivity of the test and this also helped in reducing the number of unnecessary biopsies. In 103 men who were biopsied, 13 (12.6%) prostatic carcinoma were identified. Among these 13 cancer patients, 9 patients had abnormal findings in DRE.7 individuals out of these 9, also had free to total PSA ratio lower than 0.16 and would have been biopsied and diagnosed anyway. If we use only f/t PSA ratio less than 0.16, to decide whom to biopsy, we would have biopsied and diagnosed 11/13 cases i.e. sensitivity of 85% but If we decide to biopsy those patients who had abnormal DRE and those who had low f/t PSA ratio, we could identify 13/13 carcinoma i.e. 100% sensitivity.

Combining the f/t PSA ratio with total PSA, DRE and TRUS findings could help in reducing the number of unnecessary biopsies. 37 patients who were negative for malignancy having total PSA in the range of 5–20 ng/ml, normal DRE and TRUS findings, have been biopsied but with combination of total PSA in the range of 5–20 ng/ml, normal findings in digital rectal examination and TRUS and f/t PSA ratio more than 0.16 (cutoff), we could have avoided 16 biopsies which were unnecessary that means there was 43% reduction in unnecessary biopsies.
Heart Disease, Clinical Proteomics and Mass Spectrometry
Authors: Stanley Brian A. , Gundry Rebekah L. , Cotter Robert J. , Van Eyk Jennifer E.
Published: June 2013

Heart disease is the leading cause of mortality and morbidity in the world. As such, biomarkers are needed for the diagnosis, prognosis, therapeutic monitoring and risk stratification of acute injury (acute myocardial infarction (AMI)) and chronic disease (heart failure). The procedure for biomarker development involves the discovery, validation, and translation into clinical practice of a panel of candidate proteins to monitor risk of heart disease. Two types of biomarkers are possible; heart‐specific and cardiovascular pulmonary system monitoring markers. Here we review the use of MS in the process of cardiac biomarker discovery and validation by proteomic analysis of cardiac myocytes/tissue or serum/plasma. An example of the use of MS in biomarker discovery is given in which the albumin binding protein sub‐proteome was examined using MALDI‐TOF MS/MS. Additionally, an example of MS in protein validation is given using affinity surface enhanced laser desorption ionization (SELDI) to monitor the disease‐induced post‐translational modification and the ternary status of myoctye‐originating protein, cardiac troponin I in serum.
Mutations in Transglutaminase 1 Gene in Autosomal Recessive Congenital Ichthyosis in Egyptian Families
Authors: Shawky R. M. , Sayed N.S. , Elhawary N.A.
Published: June 2013

Autosomal recessive congenital ichthyosis (ARCI) is a rare heterogeneous keratinization disorder of the skin. It is clinically divided into 2 subtypes, lamellar ichthyosis (LI) and congenital ichthyosiformis erythroderma (CIE). We investigated forty‐three ARCI Egyptian individuals in 16 severe LI, and 10 CIE families. We identified 5 alleles in two Egyptian families as having intron‐5/exon‐6 splice acceptor mutation recognized by the MspI restriction endonuclease. This promoted to a frequency of 9.6% for this mutation (5 splice‐mutation alleles/52 alleles tested). We extended our previous dataset to update the detection of R142H mutation in 4 CIE Egyptian families and one LI phenotype (frequency of 28.8%; 15/52), whereas we still had no R141H among our Egyptian population. There was no correlation between phenotype and genotype in our study. Surprisingly, the mutant alleles detected in intron‐5 acceptor splice‐site were associated with the other extreme of CIE phenotypes rather than the severe LI form. We clearly demonstrated that the ARCI Egyptian families in Upper Egypt was ethnically pure and had a tendency not to be a hybrid with other populations in Lower Egypt, Delta zone and Cairo city.
Matrix Metalloproteinase‐2, Squamous Cell Carcinoma Antigen, and Tissue Polypeptide‐Specific Antigen Expression in Egyptian Patients with Cervical Carcinoma: Relationship with Prognosis
Authors: Ahmed Maha Imam , Salahy Eman-El , Tawfiq Hassan , Khalifa Ali , Hassan Manal M.
Published: June 2013

Matrix metalloproteinases (MMPs), a family of proteolytic enzymes produced by both stromal and tumor cells, appear to have a key role in the events leading to local invasion and metastasis by malignant neoplasms. In the present study, we evaluated the role of MMP‐2, squamous cell carcinoma antigen (SCCA), and tissue polypeptide – specific antigen (TPS) in cervical neoplasia. Using Western blotting and enzyme immunoassay (EIA), we analyzed 50 patients with cervical carcinoma (CC) and 25 normal controls for expression of MMP‐2 in tissue cell lysates. We also quantified SCCA and TPS with microparticle immunoassay and EIA, respectively. The results were correlated with human papilloma virus (HPV) infection, clinicopathological findings, and disease outcome. The cutoff point for each marker was estimated from receiver operating characteristic curves. Logistic regression analysis was performed to estimate the odds ratio (OR) and 95% confidence interval (CI) for each marker. MMP‐2, SCCA, and TPS protein expression were significantly higher in patients with CC than in normal controls. While TPS was the best marker for discriminating between patients and controls, MMP‐2 was associated with an advanced tumor stage (OR, 13.9 [95% CI, 1.4‐133.9]) and poor histological grade (OR, 10.2 [95% CI, 1.7‐60.5]). Moreover, independent of the effect of an advanced CC stage and grade, the patients′ age, and the presence of HPV infection, MMP‐2 was considered a strong predictor for CC recurrence (OR, 8.1 [95% CI, 1.3‐ 49.1]). Tissue markers may be used to select high‐risk patients for early detection of and adjuvant therapy for recurrence. Our MMP‐2 findings are particularly relevant to the development of protease inhibitors as a new cancer therapy approach.
Circulating Nucleic Acids in Plasma or Serum (CNAPS) as Prognostic and Predictive Markers in Patients with Solid Neoplasias
Authors: Goebel Georg , Zitt Marion , Zitt Matthias , Müller Hannes M.
Published: June 2013

It is now widely accepted that there is a need for the development of molecular markers of cancer that can be used for clinical prognostication and monitoring. Approximately a decade ago tumor‐derived circulating nucleic acids in the plasma or serum (CNAPS) of cancer patients were introduced as a noninvasive tool for cancer detection. This review focuses on the various types of CNAPS of patients with solid neoplasias (genetic alterations in circulating DNA, microsatellites, methylated DNA, viral DNA, nucleosomes, mitochondrial DNA and cell‐free mRNA) and their putative potential as prognostic or predictive parameter or even as a tool for therapy monitoring during follow‐up. Additionally, this review aims to point out the difference between a prognostic and a predictive factor in patient bloodstream. However, with rapid technical improvement and well‐designed studies we conclude that the next years will see CNAPS analysis integrated in the prognostication and monitoring of cancer patients, thus producing more specific treatment regimens for patients with various stages of neoplastic disease and ultimately longer survival and better quality of life.
MUC Gene Abnormalities in Sporadic and Hereditary Mucinous Colon Cancers with Microsatellite Instability
Authors: Pastrello Chiara , Santarosa Manuela , Fornasarig Mara , Sigon Roberto , Perin Tiziana , Giannini Giuseppe , Boiocchi Mauro , Viel Alessandra
Published: June 2013

Aim of this study was verifying whether mucin producing colon cancers (CRCs) could develop through a molecular pathway involving microsatellite instability (MSI) and MUC gene alterations. Out of 49 CRCs expressing variable amounts of mucin, 22 (44.9%) were MSI‐H and 5 (10.2%) were MSI‐L. MUC genes were analyzed by Southern blotting and extra bands were evident in the Variable Number Tandem Repetition (VNTR) regions of MUC2 (5 cases) and MUC5AC (2 cases), but not MUC1 and MUC4 genes. Since the somatic VNTR abnormalities were detected in 6 MSI‐H and in 1 MSI‐L tumors, they seem to be peculiar of mismatch repair defective CRCs. Our finding suggests that alteration and/or loss of structurally normal MUC genes may be an important step in the neoplastic molecular pathway of a subset of CRCs and that mutations involving VNTR repetitive sequences may exist in MSI tumors as a direct and/or indirect consequence of an inefficient MMR system.
The Gly482Ser Missense Mutation of the Peroxisome Proliferator-Activated Receptor γ Coactivator‐1α (PGC‐1α) Gene Associates with Reduced Insulin Sensitivity in Normal and Glucose‐Intolerant Obese Subjects
Authors: Fanelli Marzia , Filippi Emanuela , Sentinelli Federica , Romeo Stefano , Fallarino Mara , Buzzetti Raffaella , Leonetti Frida , Baroni Marco Giorgio
Published: June 2013

Among the putative candidate genes for insulin resistance, the peroxisome proliferator‐activated receptor γ coactivator‐1α (PGC‐1α) is a transcriptional coactivator of PPARγ and α, regulating a wide range of processes involved in energy production and utilization, such as thermogenesis, liver gluconeogenesis, glucose uptake in muscle. In population studies a Gly482Ser substitution in PGC‐1α has been reported to be associated with increased risk of type diabetes 2 and insulin resistance. In the present study we have analysed the association between the Gly482Ser missense mutation of the PGC‐1α gene and insulin sensitivity and glucose tolerance in a population of obese non‐diabetic subjects. The Gly482Ser SNPs was detected by PCR‐RFLP in a cohort of 358 Caucasian obese subjects (223 with normal glucose tolerance (NGT) and 125 with impaired glucose tolerance (IGT). We observed a significant association (p < 0.007) between carriers of the Gly482Ser variant of the PGC‐1α gene and insulin resistance measured by HOMAIR. Multivariate analysis confirmed that the Gly482Ser SNP was a significant (p < 0.02) determinant of decreased insulin sensitivity, independently from other well‐known modulators of insulin action. In conclusion, we have found significant association between the Gly482Ser variant of the PGC‐1α gene and reduced insulin sensitivity in obese subjects. This association resulted independent from all other known modulators of insulin resistance, and suggests a primary role for the PGC‐1α gene on the genetic susceptibility to insulin resistance in obesity.
Salivary Thiobarbituric Acid Reacting Substances and Malondialdehyde – Their Relationship to Reported Smoking and to Parodontal Status Described by the Papillary bleeding index
Authors: Celec Peter , Hodosy Július , Celecová Viera , Vodrážka Ján , Červenka Tomáš , Halčák Lukáč , Božek Peter , Kopáni Martin , Kúdela Matúš
Published: June 2013

Background. Thiobarbituric reacting substances (TBARS) are markers of lipoperoxidation. The best‐known specific TBARS is malondialdehyde (MDA). Results from our previous studies have shown that TBARS can be measured in saliva and are increased in patients with gingivitis. Whether MDA is the main TBARS in saliva from patients with altered parodontal status is unknown. Aim. To observe the relationship between the parodontal status and TBARS, MDA and the number of epithelial cells in saliva. Subjects & Methods. In Study I saliva and plasma samples of 15 patients (8F, 7M) suffering from inflammatory periodontal diseases were gathered and TBARS levels were measured in these samples. In Study II saliva samples from 217 consecutive stomatologic patients were collected and analysed for TBARS spectrofluorometrically, MDA by high‐performance liquid chromatography and epithelial cell count by light microscopy. Papillary bleeding index (PBI) was determined in standard stomatologic examination. Results. In Study I results from our previous studies showing no correlation between salivary and plasma TBARS levels were confirmed. This indicates that the local salivary level of TBARS is unlikely to be directly affected by systemic oxidative stress. In Study II higher PBI was associated independently (adjusted for age and sex) tightly with higher TBARS (p < 0.001) and with
lower number of epithelial cells in saliva (p < 0.05). Smokers had higher salivary MDA levels (p < 0.003) and lower number
of epithelial cells in saliva (p < 0.01). Conclusion. Salivary TBARS are a simple parameter that partially reflects the parodontal status with a potential usefulness in the clinical stomatology. We show herein that salivary MDA is dependent on age and smoking, but there is no correlation between MDA and PBI. Further studies should uncover the main salivary TBARS compound in patients with altered parodontal status and trace the origin of these salivary lipoperoxidation markers.
TNF‐α, TNF‐β, IL‐6, and IL‐10 Polymorphisms in Patients with Lung Cancer
Authors: Seifart Carola , Plagens Alexandra , Dempfle Astrid , Clostermann Ursula , Vogelmeier Claus , von Wichert Peter , Seifart Ulf
Published: June 2013

Apart from cigarette smoking, genetic factors seem to be of importance in the development of lung cancer. The present case‐control study investigated frequencies of five inflammatory response gene polymorphisms (TNF‐α‐308, TNF‐β‐Intron1‐252, IL‐6‐174, IL‐10‐819 and IL‐10‐1082) in patients with lung cancer and controls. The study population consisted of 117 patients with lung cancer (77 patients with NSCLC, including 40 Squamous Cell Carcinoma and 26 Adenocarcinoma, and 40 patients with SCLC), 117 matched controls without pulmonary disease and 243 healthy individuals (population control). Genotype analyses revealed no difference in genotype frequencies using matched‐pair analysis. However, in comparison to the population control, the IL‐10‐1082 genotypes carrying the G allele appeared with higher frequency in the SCLC group (p = 0.006) [SCLC: 84.6%, population controls: 64.6%]. This yields an odds ratio of 3.01 for SCLC (95% CI = [1.21, 7.48]). No associations were seen for all other polymorphisms analysed. The study raises the possibility of a correlation between the IL‐10‐1082_G allele and the presence of SCLC in a German population. The functional IL‐10‐1082 polymorphism correlates with altered IL‐10 levels and might influence lung cancer susceptibility by altered inflammatory responses in the airways.
Two Polymorphisms in the Epithelial Cell‐Derived Neutrophil‐Activating Peptide (ENA‐78) Gene
Authors: Amoli Mahsa M. , Larijani Bagher , Thomson Wendy , Ollier William E. R. , Gonzalez-Gay Miguel A.
Published: June 2013

Increased expression of epithelial cell‐derived neutrophil‐activating peptide (ENA‐78) has been reported in several immune and inflammatory conditions suggesting its role in inflammatory response. We have identified two single nucleotide polymorphisms in the promoter and exon 2 of the ENA‐78 gene by scanning the full length gene using DHPLC DNA fragment analysis and DNA sequencing.

The polymorphism at position +398 (A/G from the first ATG codon) in exon 2 results in a synonymous substitution not resulting in an amino acid change. The promoter polymorphism was found at position −156 (C/G from the first ATG codon). An assay was designed for the detection of the polymorphisms using SNapshot ddNTP primer extension, followed by capillary electrophoresis (ABI 3100).

Allele and genotype frequencies for the promoter −156 polymorphism are presented for 107 healthy Spanish and 54 UK Caucasians. Frequencies for the exon 2 polymorphism are also presented for 63 UK Caucasians.
Altered β‐Amyloid Precursor Protein Isoforms in Mexican Alzheimer’s Disease Patients
Authors: Sánchez-González V. J. , Ortiz G. G. , Gallegos-Arreola P. , Macías-Islas M. A. , Arias-Merino E. D. , Loera-Castañeda V. , Martínez-Cano E. , Velázquez-Brizuela I. E. , Rosales-Corral S. A. , Curiel-Ortega C. R. , Pacheco-Moisés F. , García J. J.
Published: June 2013

Objective: To determine the β‐amyloid precursor protein (βAPP) isoforms ratio as a risk factor for Alzheimer’s Disease and to assess its relationship with demographic and genetic variables of the disease.

Methods: Blood samples from 26 patients fulfilling NINCDS‐ADRDA diagnostic criteria for AD and 46 healthy control subjects were collected for Western blotting for βAPP. A ratio of βAPP isoforms, in optical densities, between the upper band (130 Kd) and the lower bands (106–110 Kd) was obtained. Odds ratios were obtained to determine risk factor of this component.

Results: βAPP ratio on AD subjects was lower than that of control subjects: 0.3662 ± 0.1891 vs. 0.6769 ± 0.1021 (mean ± SD, p<0.05). A low βAPP ratio (<0.6) showed an OR of 4.63 (95% CI 1.45 ± 15.33). When onset of disease was taken into account, a βAPP ratio on EOAD subjects of 0.3965 ± 0.1916 was found vs. 0.3445 ± 0.1965 on LOAD subjects (p>0.05).

Conclusions: Altered βAPP isoforms is a high risk factor for Alzheimer’s disease, although it has no influence on the time of onset of the disease.
Genetic Polymorphisms of Phase II Metabolic Enzymes and Lung Cancer Susceptibility in a Population of Central South China
Authors: Chen Han-chun , Cao Yan-fei , Hu Wei-xin , Liu Xin-fa , Liu Qing-xia , Zhang Ji , Liu Jia
Published: June 2013

A case‐control study was conducted for analyzing the genetic polymorphisms of phase II metabolic enzymes in 97 patients with lung cancer and 197 healthy subjects from Han ethnic group of Hunan Province located in Central South China. The results showed that the frequencies of glutathione S‐transferase (GST) M1‐null (GSTM1‐) or GSTT1‐null (GSTT1‐) genotype alone, or combined form of both in lung cancer patients were significantly higher than those of the controls. Genotypes of combining GSTP1 mutant/GSTM1(‐) or GSTP1 mutant/GSTT1(‐) led to high risk of lung cancer. Individuals carrying any two or all three of GSTM1(‐), GSTT1(‐) and GSTP1 mutant genotypes have a distinctly increased risk of lung cancer when compared to those with GSTM1 present (GSTM1+: GSTM1+/+ or GSTM1+/−), GSTT1 present (GSTT1+: GSTT1+/+ or GSTT1+/−) and GSTP1 wild genotypes. Furthermore, individuals possessing combined genotypes of N‐acetyltransferase 2 (NAT2) rapid acetylator, GSTP1 mutant and both GSTT1(‐) and GSTM1(‐) have a remarkably higher lung cancer risk than those carrying combined NAT2 slow acetylator genotype, GSTP1 wild genotype and both GSTT1(+) and GSTM1(+) genotypes. All these findings suggest that the genetic polymorphisms of phase II metabolic enzymes affect the susceptibility of lung cancer in the Han ethnic group of Central South China.