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      Detection of Toxoplasma gondii DNA in Malignant Breast Tissues in Breast Cancer Patients

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          Abstract

          Breast cancer is the most prevalent malignancy in women throughout the world. Similar to other cancers, a strong relationship between breast cancer and environmental factors such as infectious agents has been reported. Toxoplasma gondii is a protozoan parasite which may play a role in cancer induction. The present study aimed to investigate a possible association between a history of T. gondii infection and breast cancer by detecting T. gondii DNA in malignant and non-malignant breast and lymph nodes tissues from breast cancer patients with latent toxoplasmosis. Formalin-fixed, paraffin-embedded (FFPE) tissue blocks from malignant/non-malignant breast and lymph nodes were obtained from twenty-nine breast cancer patients who were positive for anti- Toxoplasma antibodies (IgG). FFPE tissue blocks were deparaffinized using hot water method, and DNA was extracted. A conventional PCR analysis was performed to amplify partial regions of T. gondii B1 and REP-529 genes. Ninety-three samples from 29 patients were examined. All patients were negative for anti- T. gondii antibodies (IgM). T. gondii DNA was detected in 3 (10.3%) patients by PCR analysis of either B1 or REP-529 genes. These include two malignant breast and one normal lymph node samples. Sequence analysis of these genes showed a good similarity with previously published B1 and REP-529 sequences of T. gondii in NCBI GenBank. This study did not find any association between T. gondii infection and breast cancer. Furthermore, it is the first molecular identification of T. gondii in FFPE tissue samples obtained from breast cancer patients.

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          Most cited references18

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          Genetic and hormonal risk factors in breast cancer.

          Breast cancer poses a serious public health problem, and it is hoped that identification of genetic and environmental factors that contribute to the development of breast cancer will enhance prevention efforts. Two breast cancer susceptibility genes (BRCA1 and BRCA2) have been identified, and germline mutations in these genes are thought to account for between 5% and 10% of all breast cancer cases. Current findings suggest that mutations in other highly penetrant genes may play an important role in breast cancer susceptibility, and studies aimed at the isolation of these genes are under way. In addition, common variants in a number of gene classes are thought to act as low-penetrance susceptibility alleles, and efforts to identify and characterize these variants are under way. This review discusses the genetic components of susceptibility to breast cancer from the standpoint of both human genetics and rat models.
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            Efficient DNA extraction for HPV genotyping in formalin-fixed, paraffin-embedded tissues.

            DNA from archived FFPE can be used for papillomavirus genotyping, but potential problems include paraffin as a physical barrier, DNA cross-linking, and PCR inhibitors. To address these complications, we combined a commercially available DNA isolation kit (Qiagen DNeasy) with a heat treatment and evaluated the resulting DNA with regards to HPV typing. DNA was extracted from 10-μm sections from 150 FFPE cancer samples. One protocol followed the manufacturer's recommendation, including paraffin removal by xylene and tissue lysis at 56°C. A second section was directly incubated at 120°C and subsequently lysed at 65°C. After spin-column purification, both extracts were tested with a linear array HPV genotyping assay. Additionally, cellular DNA yield, HPV16 DNA copies, and PCR inhibitors were assessed by real-time qPCR assays. Inadequate linear array HPV genotyping assay results were significantly more frequent (P = 0.0003) in xylene-treated (29/150, 19.3%) than in heat-treated extracts (8/150, 5.3%). HPV detection also differed, with 94/150 (62.7%) and 110/150 (73.3%) positive results, respectively (P = 0.0026). The heat method also yielded more PCR-amplifiable cellular DNA (8.2-fold; P < 0.001) and HPV16 copies (6.5-fold; P = 0.009), although PCR inhibitors also had a greater effect (P = 0.035). Aggressive heat treatment demonstrated an advantage over traditional xylene purification protocols, resulting in higher DNA yields and increased sensitivity for HPV testing. Published by Elsevier Inc.
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              Brain cancer mortality rates increase with Toxoplasma gondii seroprevalence in France.

              The incidence of adult brain cancer was previously shown to be higher in countries where the parasite Toxoplasma gondii is common, suggesting that this brain protozoan could potentially increase the risk of tumor formation. Using countries as replicates has, however, several potential confounding factors, particularly because detection rates vary with country wealth. Using an independent dataset entirely within France, we further establish the significance of the association between T. gondii and brain cancer and find additional demographic resolution. In adult age classes 55 years and older, regional mortality rates due to brain cancer correlated positively with the local seroprevalence of T. gondii. This effect was particularly strong for men. While this novel evidence of a significant statistical association between T. gondii infection and brain cancer does not demonstrate causation, these results suggest that investigations at the scale of the individual are merited. Copyright © 2012 Elsevier B.V. All rights reserved.
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                Author and article information

                Journal
                Int J Mol Cell Med
                Int J Mol Cell Med
                IJMCM
                International Journal of Molecular and Cellular Medicine
                Babol University of Medical Sciences (Babol, Iran )
                2251-9637
                2251-9645
                Summer 2017
                26 September 2017
                : 6
                : 3
                : 190-196
                Affiliations
                [1 ] Cellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences, Babol, Iran.
                [2 ] Infectious Diseases and Tropical Medicine Research Center, Babol University of Medical Sciences, Babol, Iran.
                [3 ] Department of Pathology, Faculty of Medicine, Babol University of Medical Sciences, Babol, Iran.
                [4 ] Department of Surgery, Cancer Research Center, Babol University of Medical Sciences, Babol, Iran.
                [5 ] Student Research Committee, Babol University of Medical Sciences, Babol, Iran.
                [6 ] Department of Parasitology and Mycology, Faculty of Medicine, Babol University of Medical Sciences, Babol, Iran.
                [# ]First two authors have equal contribution.
                Author notes
                [* ]Corresponding author: Infectious Diseases and Tropical Medicine Research Center, Babol University of Medical Sciences, Babol, Iran. Email: m_baiany@yahoo.com
                Article
                10.22088/acadpub.BUMS.6.3.190
                5898643
                29682491
                69292b88-4eef-44bd-9086-980a759eeb5a

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License, ( http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 14 June 2017
                : 12 September 2017
                Categories
                Short Communication

                breast cancer,ffpe tissues,pcr,t. gondii,women
                breast cancer, ffpe tissues, pcr, t. gondii, women

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