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జర్నల్ ఆఫ్ మాలిక్యులర్ బయోమార్కర్స్ & డయాగ్నోసిస్

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వాల్యూమ్ 5, సమస్య 2 (2014)

పరిశోధన వ్యాసం

PON I and GLO I Gene Polymorphisms and Their Association with Breast Cancer: A Case-Control Study in a Population from Southern Italy

Carmela Rinaldi, Rosalia D'Angelo, Alessia Ruggeri, Marco Calabro, Concetta Scimone and Antonina Sidoti

Background: Many factors may play a role in the susceptibility to the breast cancer. Oxidative stress may be one of these. Polymorphisms of genes such as paraoxonase I (PON I) and glyoxalase I (GLO I) may influence individual susceptibility to breast cancer. In the present study, we have conducted a case-control study in order to examine the possible relation between GLO I A111E and PON I Q192R/L55M polymorphisms with the risk of breast cancer.

Methods: The three polymorphisms were characterized in 144 breast cancer postmenopausal patients and in 152 healthy women by PCR/RFLP methods using DNA from lymphocytes.

Results: Among the three polymorphisms, only PON I L55M polymorphism was associated with the patient’s age and, more precisely, the heterozygous genotype that is more represented in women aged between 51-69 years. In addition, we found that individuals with the PON192 Q/R - R/R genotypes and PON55 L/M - M/M genotypes had a significantly higher risk of breast cancer compared with the other genotypes. The genotypes PON55 L/M and PON192 Q/R showed significant association with lymph nodes positivity (p < 0.001) and with a high nuclear grading (p < 0.001), respectively. Conversely the genotypes GLO I AE/EE were associated with a low nuclear grading.

Conclusions: We believe that the combination of the three polymorphisms may be a more predictive factor for the risk of this neoplasia in each single examined case.

పరిశోధన వ్యాసం

Molecular Characterization of Trichodermaviride Isolated from Rhizospheric Soils of Uttar Pradesh Based on rDNA Markers and Analysis of Their PCR-ISSR Profiles

Mohammad Shahid, Anuradha Singh, Mukesh Srivastava and Srivastava DK

The objectives of this research were to characterize isolates of Trichoderma collected from rhizospheres of chickpea, pigeonpea and lentil crop from different places of Uttar Pradesh, India, using microsatellite-primed polymerase chain reaction (MP-PCR) and ribosomal DNA (rDNA) sequence analysis and to combine these results with morphological characteristics for classification. Thirty isolates of Trichodermaviride obtained from rhizosphere soil of plantation crops, and agricultural fields of UP region were studied using ISSR and ITS-PCR. The genetic relatedness among thirty isolates of T. viride were analyzed with six microsatellite primers. ISSR profiles showed genetic diversity among the isolates with the formation of eight clusters. Analysis of dendrogram revealed that similarity coefficient ranged from 0.27 to 0.95. ITS-PCR of rDNA region with ITS1 and ITS4 primers produced 600bp products in all isolates. This result indicated the identification patterns of Trichoderma isolates.

పరిశోధన వ్యాసం

Novel Mutation in the CFTR Gene of Cystic Fibrosis Patients in Oman

Hussein Al-Kindy, Allal Ouhtit, Qasim Al-Salmi, Muna Al-Bimani, Mariam Al-Nabhani and Ishita Gupta

Introduction: Cystic fibrosis (CF) is the most common lethal autosomal recessive disorder among Caucasians (1: 3,000). In CF, the CFTR gene is frequently mutated, with ΔF508del being the largely common mutation in Caucasians. Our preliminary pilot study in Omani population revealed a CF prevalence of 1:2,738.

Objective: The objective of the present study was to determine the most common CFTR mutations in the Omani patients to establish a proper molecular genetics diagnostic basis of CF in Oman. Methods: Blood Genomic DNA samples from Omani patients were examined by PCR and sequencing analyses for the entire coding sequence of the CFTR gene were performed.

Results: The innovative aspect of this study was the identification of a novel CF-causing mutation, L578delTA. Furthermore, in contrast to the west, p.S549R appears to be the most common mutation in Oman (65.2% for S549R and 13% for ΔF508).

Conclusion: The mutation spectrum of CF in Oman revealed six CF-causing mutations, p.S549R, ΔF508, 3120+1G>A, L578delTA, p.A357T and 3849+10kbC->T. These findings will ultimately pave the way towards the development of molecular genetic tests in Oman to confirm the diagnosis of CF and serve the patient care and management.

పరిశోధన వ్యాసం

Increased Serum Hepatocyte Growth Factor (HGF) Levels in Patients with Idiopathic Pulmonary Fibrosis (IPF) or Progressive Sarcoidosis

Dariusz Ziora, Mariusz Adamek, Zenon Czuba, Dariusz Jastrzębski, Karolina Zeleznik, Slawomir Kasperczyk, Jerzy Kozielski and Wojciech Kro

Background: Endogenous Hepatocyte Growth Factor (HGF) is required for self-repair of injured liver, kidneys and lungs. It also exerts a regenerative effect on epithelium. HGF can inhibit both the initiation and progression of lung fibrosis in Idiopathic Pulmonary Fibrosis (IPF). However available literature data on the role of HGF in sarcoidosis are scarce and conflicting. The aim of our study was to estimate and compare the serum concentrations of HGF both in patients with IPF and sarcoidosis and in healthy controls.

Material and methods: In 43 patients with sarcoidosis, 17 patients with IPF and 20 healthy controls serum HGF concentrations were measured using multiplex bead based sandwich immunoassay.

Results: In patients with IPF mean concentrations of HGF (1286.2 ± 238.3 pg/ml) were significantly higher than in controls (819.8 ± 61.0 pg/ml, p<0.05), but not significantly higher than in sarcoidosis patients (990.0 ± 52.2 pg/ ml). In all patients with sarcoidosis the mean levels of HGF did not differ significantly from levels obtained in control group. However, the mean levels of HGF in 18 patients with progressive sarcoidosis (1260.7 ± 83.7 pg/ml) were significantly higher (p<0.001) than in 25 patients with non-active, stable sarcoidosis (802.7 ± 34.8 pg/ml) and controls (p<0.05).

Conclusion: The role of serum HGF as a potential biomarker in sarcoidosis needs further explorations to answer the question if increased HGF serum levels can predict deterioration of the disease or reflect lung epithelial injury by granulomatous inflammation.

పరిశోధన వ్యాసం

The Effect of Amino-Bisphosphonate Therapy on Gingival Crevicular Fluid Alkaline Phosphatase Activity in Cancer Patients

Katia Rupel, Giuseppe Perinetti, Luca Contardo, Giulia Ottaviani, Margherita Gobbo, Sara De Iudicibus, Roberto Di Lenarda and Matteo Biasotto

The enzyme Alkaline Phosphatase (ALP) is considered a biomarker of bone formation and it is used for monitoring intravenous Bisphosphonate (BP) therapy. It can be measured in serum and urine, but also in Gingival Crevicular Fluid (GCF). The aim of the present study was to monitor the GCF-ALP activity fluctuations in cancer patients before and during the first six months of BP therapy, comparing them with systemic serum ALP concentrations. 33 patients and 25 controls were enrolled according to the eligibility criteria. The GCF was collected at baseline, and after every BP administration for 6 times. The total ALP activities were determined spectrophotometrically. The results show how the differences in GCF-ALP activity over time appear to be significant in the BP-taking group (p=0.003), while the differences in serum ALP in the BP-taking group and in GCF-ALP activity in the control group were not significant (p>0.05). GCF-ALP activity may thus represent a possible new biomarker for bone turnover rate evaluation in patients taking intravenous BP therapy. Up to date, this is the first study that investigates on variations of GCF-ALP activity due to changes in bone metabolism caused by intravenous nitrogen-containing BP therapy in cancer patients.

పరిశోధన వ్యాసం

Feasibility of Protein Biomarkers in the Prediction of Subclinical Doxorubicin Nephrotoxicity in Male Sprague-Dawley Rat

J. Eric McDuffie, Manisha Sonee, Jing Ma, Frederic Almy, Xuejun Liu, David La and Sandra Snook

The feasibility of proteinbiomarkers in the prediction of subclinical doxorubicin nephrotoxicity was evaluated in male Sprague-Dawley rats during a 2-week study with once‑weekly dosing. Doxorubicin (5, 7.5, and 10 mg/kg/ dose) or 0.9% Saline was intravenously administered on days 1 and 8. Urine and serum were collected at various time points. Surviving animals were euthanized on day 14, and tissues were collected for microscopic examination. Severe clinical signs were observed in the 7.5 and 10 mg/kg/dose groups. Biomarker data are not reported for these groups because the objective of this study was to evaluate biomarkers at doses not associated with clinical signs. In the 5 mg/kg group, increased serum concentrations of urea nitrogen were observed on day 14 with concurrent renal histopathology findings which were primarily characterized as slight renal glomerular and tubular injury with mild multi-focal intratubular hyaline casts consistent with protein leakage from damaged glomeruli. Of the various urinary protein biomarkers examined, increased urinary concentrations of albumin was observed on day 7 and increased total protein, albumin and lipocalin‑2 were observed on day 14. Taken together, these findings showed that urinary albumin was more sensitive and selective than urinary total protein, lipocalin-2, kidney injury molecule 1 and/or osteopontin in the prediction of progressive doxorubicin‑induced glomerular toxicity with secondary renal tubular toxicity in male Sprague‑Dawley rats.

పరిశోధన వ్యాసం

Expression of Complement Receptor Type 1 on Erythrocytes in Autoimmune Diseases

Deng-Ho Yang, Chen-Hung Chen, Cheng-Chung Wei and Ya-Wen Cheng

Objective: The expression of complement receptor type 1 on different cells is associated with autoimmunity. Erythrocyte-Complement Receptor Type 1 (E-CR1) is a candidate for early diagnosis of Systemic Lupus Erythematosus (SLE) and assessed the roles of disease activity. We evaluated the expression of E-CR1 in the patients with SLE and other autoimmune diseases.

Methods: We conducted a cross-sectional investigation of 3 groups: (1) 36 patients with SLE; (2) 51 patients with other diseases including Rheumatoid Arthritis (RA), Sjögren’s syndrome, anti-phospholipid syndrome, Mixed Connective Tissue Disease (MCTD), Raynaud’s disease, ankylosing spondylitis, pulmonary Tuberculosis (TB); and (3) 26 healthy controls. Erythrocytes were analyzed by flow cytometry to determine levels of E-CR1.

Results: We found a significant reduction in the mean levels of E-CR1 in SLE patients compared to patients with healthy controls (1.79 ± 0.16 versus 3.82 ± 0.32, P<0.05). However, in patients with RA, MCTD, and TB, decreased E-CR1 levels were also observed. There was a statistically significant correlation between reduced levels of E-CR1 and SLE disease activity index (r=-0.326, P<0.05). The E-CR1 levels were inversely correlated with urine daily protein loss (r=-0.364, P<0.05).

Conclusions: Although E-CR1 levels may be a useful diagnostic marker for SLE, a possible limitation is that no significant differences in E-CR1 levels were observed among patients with SLE, those with MCTD and TB. Overall, E-CR1 levels in SLE patients are related to disease activity index and reflect renal clearance through urine daily protein loss.

పరిశోధన వ్యాసం

Lipocalin 2 Level Predicts Prognostic Outcome in Patients with Colorectal Cancer Undergoing Surgical Intervention

Chia-Lo Chang, Yung-Lung Chen, Yen-Ta Chen, Steve Leu, Li-Teh Chang, Chang-Han Chen, Chien-Chang Lu, Wang-Hseng Hu, Ko- Chao Lee, Kai-Lung Tsai, Hsueh-Wen Chang, Hong-Hwa Chen and Hon-Kan Yip

This study examined the predictive role of circulating lipocalin 2 in long-term prognostic outcome of Colorectal Cancer (CRC) patients with surgical intervention. 204 CRC patients with surgical intervention between April 2009 and May 2011 were prospectively enrolled. Circulating lipocalin 2 levels in CRC patients before surgery and healthy-control subjects were measured using ELISA. Patients were categorized into different CRC clinical stage (I to IV) based on pathological findings. Mean follow-up time was 21.2 ± 3.5 months. Patients were categorized into group 1 (n=67), lipocalin 2 levels <116.7 ng/ ml; and group 2 (n=137), lipocalin 2 levels ≥ 116.7 ng/ml by ROC curve analysis. CRC patients had higher circulating levels of lipocalin 2 than healthy subjects (P<0.0001). Occurrence of new metastases after surgery, primary (any cause of death or new metastasis) and secondary (CRC-related death and new metastasis) combined endpoint were higher in group 1 than group 2 (all P<0.03). Lower lipocalin 2 levels and CRC stage IV was independently predictive of primary and secondary combined endpoint (all P<0.001) during long-term follow-up. In conclusion, circulating lipocalin 2 level may be a useful biomarker for risk stratification of CRC patients during long-term follow-up.

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