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Rebecca J Stoll, Grace R Thompson, Mohammad D Samy, George Blanck
Human mutagenesis is largely random, thus large coding regions, simply on the basis of probability, represent relatively large mutagenesis targets. Thus, we considered the possibility that large cytoskeletal-protein related coding regions (CPCRs), including extra-cellular matrix (ECM) coding regions, would have systemic nucleotide variants that are not present in common SNP databases. Presumably, such variants arose recently in development or in recent, preceding generations. Using matched breast cancer and blood-derived normal datasets from the cancer genome atlas, CPCR single nucleotide variants (SNVs) not present in the All SNPs(142) or 1000 Genomes databases were identified...
February 2017: Biomedical Reports
Pavlo Maksimov, Gereon Schares, Sebastian Press, Andreas Fröhlich, Walter Basso, Mandy Herzig, Franz J Conraths
Effective and sensitive methods for the molecular detection of Echinococcus multilocularis in faecal samples of final hosts are crucial for the prevention and control of human alveolar echinococcosis and for studies on the epidemiology of the parasite. Little is known about the suitability of commercial test kits for isolating DNA of E. multilocularis from fox faeces and the performance of standard Polymerase Chain Reaction (PCR) protocols in relation to the quality of DNA extracted by these kits. We compared four different kits: ZR Faecal DNA MiniPrep™ (Zymo Research), FastDNA(®) SPIN Kit for Soil (MP Biomedicals), QIAamp(®) Fast DNA Stool Mini Kit (QIAGEN) and NucleoSpin(®) Soil Kit (Macherey-Nagel) for the extraction of DNA from E...
February 24, 2017: Veterinary Parasitology
Sharon C Clouthier, Carol McClure, Tamara Schroeder, Megan Desai, Laura Hawley, Sunita Khatkar, Melissa Lindsay, Geoff Lowe, Jon Richard, Eric D Anderson
Cyprinid herpesvirus 3 (CyHV-3) is the aetiological agent of koi herpesvirus disease in koi and common carp. The disease is notifiable to the World Organisation for Animal Health. Three tests-quantitative polymerase chain reaction (qPCR), conventional PCR (cPCR) and virus isolation by cell culture (VI)-were validated to assess their fitness as diagnostic tools for detection of CyHV-3. Test performance metrics of diagnostic accuracy were sensitivity (DSe) and specificity (DSp). Repeatability and reproducibility were measured to assess diagnostic precision...
March 6, 2017: Diseases of Aquatic Organisms
John M Yavorski, George Blanck
Cancer from smoking tobacco is considered dependent on mutagens, but significant molecular aspects of smoking-specific, cancer development remain unknown. We defined sets of coding regions for oncoproteins, tumor suppressor proteins, and cytoskeletal-related proteins that were compared between nonsmokers and smokers, for mutation occurrences, in the lung adenocarcinoma (LUAD), head and neck squamous carcinoma (HNSC), bladder carcinoma (BLCA), and pancreatic adenocarcinoma ( PAAD) datasets from the cancer genome atlas (TCGA)...
December 2016: Physiological Reports
Marwa A Ghieth, Magd A Kotb, Enas Y Abu-Sarea, Ayman A El-Badry
Giardiasis is a prevailing intestinal disease in children. This study aimed to determine molecular prevalence of Giardia intestinalis in children attending Cairo University Pediatrics Hospitals, using copro-PCR assays, conventional methods and to evaluate diagnostic effectiveness of used tests. 229 fecal samples were collected from children suffering from gastrointestinal symptoms and examined for Giardia by microscopy, Immuno-chromatographic test (ICT), copro-DNA using two PCR assays targeting tpi [nested-PCR (nPCR)] and 18S [conventional-PCR (cPCR)] genes...
December 2016: Journal of Parasitic Diseases: Official Organ of the Indian Society for Parasitology
Rodrigo C Silva, Virgínia B Richini-Pereira, Mariana Kikuti, Pâmela M Marson, Helio Langoni
BACKGROUND: Canine visceral leishmaniasis (CVL) is a worldwide parasitic zoonosis caused by Leishmania (Leishmania) infantum around the world. Canids are the definitive hosts and sand flies the intermediate hosts. OBJECTIVE: To test the hypothesis that a new species-specific primers (Lch14:Lch15, targeting a multiple alignment for L. infantum kDNA minicircle) is an efficient diagnostic tool for L. infantum. METHODS: The presence of L. infantum DNA was assessed in blood samples of 69 stray dogs using the conventional PCR (cPCR) and quantitative PCR (qPCR)...
December 2017: Veterinary Quarterly
Carolina Hernández, Zulma Cucunubá, Carolina Flórez, Mario Olivera, Carlos Valencia, Pilar Zambrano, Cielo León, Juan David Ramírez
BACKGROUND: The diagnosis of Chagas disease is complex due to the dynamics of parasitemia in the clinical phases of the disease. The molecular tests have been considered promissory because they detect the parasite in all clinical phases. Trypanosoma cruzi presents significant genetic variability and is classified into six Discrete Typing Units TcI-TcVI (DTUs) with the emergence of foreseen genotypes within TcI as TcIDom and TcI Sylvatic. The objective of this study was to determine the operating characteristics of molecular tests (conventional and Real Time PCR) for the detection of T...
September 2016: PLoS Neglected Tropical Diseases
Tegene Negesse, Chander Datt, S S Kundu
A feeding trial with 18 Murrah buffalo heifers (238 ± 63 kg; 1.7 ± 0.4 years old) was conducted for 57 days, and feed intake and body weight were recorded followed by 7 days of urine and feces collection. Heifers were grouped into low (RFI-I) and high (RFI-II) feed efficiency groups by regressing DMI (g head(-1) day(-1)) on BWG (g head(-1) day(-1)) and mid-test MBW (kg). RFI-II heifers had slightly higher DMI (g head(-1) day(-1) and expressed per unit of BW, %BW, MBW, mid-test BW, and mid-test MBW) and all associated nutrient intakes, BWG, feed efficiency traits (FCR, CPCR), digestibility of nutrients, and retentions of crude protein and fat over RFI-I heifers...
December 2016: Tropical Animal Health and Production
Candice Sant, Roger d'Abadie, Indira Pargass, Asoke K Basu, Zinora Asgarali, Roxanne A Charles, Karla C Georges
Equine piroplasmosis caused by Theileria equi and Babesia caballi is endemic in Trinidad and Tobago. Transmission occurs by ticks of the family Ixodidae. T. equi can also be transmitted transplacentally; however transplacental transmission of B. caballi is unknown. This study aims to investigate transplacental transmission of equine piroplasmosis from thoroughbred mares naturally infected via the tick vector. Whole blood and serum samples were collected from 117 mares in the fifth month of pregnancy. Blood samples were also collected from each of their foals (89 in total) within the first 36h of birth...
August 15, 2016: Veterinary Parasitology
R Ryan McNally, Carol A Ishimaru, Dean K Malvick
Goss's leaf blight and wilt of maize (corn) is a significant and reemerging disease caused by the bacterium Clavibacter michiganensis subsp. nebraskensis. Despite its importance, molecular tools for diagnosing and studying this disease remain limited. We report the identification of CMN_01184 as a novel gene target and its use in conventional PCR (cPCR) and SYBR green-based quantitative PCR (qPCR) assays for specific detection and quantification of C. michiganensis subsp. nebraskensis. The cPCR and qPCR assays based on primers targeting CMN_01184 specifically amplified only C...
October 11, 2016: Phytopathology
Shahram Bolandparvaz, Hamid Mohajer, Mansoor Masjedi, Ehsan Mohammadhoseini, Leila Shayan
OBJECTIVES: To determine the correlation between the success rates of the cardiopulmonary cerebral resuscitation (CPCR) and the team's leader education and skill level in Shiraz, southern Iran. METHOD: This cross-sectional study was conducted during a 6-month period from October 2007 to March 2008 in Nemazee hospital of Shiraz. We included all the patients who underwent CPCR due to cardiopulmonary arrest in emergency room of Nemazee hospital during the study period...
October 2015: Bulletin of Emergency and Trauma
Jinfeng Wang, Wei Ma, Yuping Xie, Peilin Hui, Lijun Zhao, Xiaoquan Wei, Liya Zhou
OBJECTIVE: To evaluate the curative effect of continuous positive airway pressure(CPAP) combined with modified oral appliance (MOA) in the treatment of severe OSAHS. METHOD: The research chose 120 male patients with severe OSAHS. They were treated with CPAP and MOA (30 cases), CPAP (30 cases), and MOA (30 cases) respectively. At the same time, the rest 30 cases without any treatment were in control group (matching factors: age, gender, IBM). After 3 months treatment, the characteristics of PSG and respiratory machine wear condition were observated in four groups...
December 2015: Journal of Clinical Otorhinolaryngology, Head, and Neck Surgery
Shinichiro Mori, Takuji Furukawa
To shorten treatment time in pencil beam scanning irradiation, we developed rapid phase-controlled rescanning (rPCR), which irradiates two or more isoenergy layers in a single gating window. Here, we evaluated carbon-ion beam dose distribution with rapid and conventional PCR (cPCR). 4 dimensional computed tomography (4DCT) imaging was performed on 12 subjects with lung or liver tumors. To compensate for intrafractional range variation, the field-specific target volume (FTV) was calculated using 4DCT within the gating window (T20-T80)...
May 21, 2016: Physics in Medicine and Biology
Jung Lo, Po-Chiung Fang, Chun-Chih Chien, Chang-Chun Hsiao, Shin-Ling Tseng, Yu-Hsuan Lai, Ming-Tse Kuo
PURPOSE: To develop a PCR gel analysis method for assessing the bacterial bioburden in orthokeratology contact lens (OK) case fluid determined by culture. METHODS: A prospective study with the participation of 41 OK wearers (20 girls, 21 boys) was performed. The mean OK-wearing experience was 3.5±1.9 years. PCR was used to assess the bacterial bioburden (colony-forming units per milliliter) of OK after removal and soaking in the storage case for 6 h. The signal intensity of the PCR bands was analyzed after grayscale image transformation...
2016: Molecular Vision
N B Vanasco, P Jacob, N Landolt, Y Chiani, M F Schmeling, C Cudos, H Tarabla, J Lottersberger
Enzyme-linked immunosorbent assay (ELISA) tests and polymerase chain reaction (PCR) may play a key role for early detection and treatment of human leptospirosis in developing countries. The aims of this study were to develop and validate an IgM ELISA under field conditions and to compare the diagnostic accuracy among IgG, IgM ELISAs, conventional PCR (cPCR), and real-time PCR (rtPCR) for early detection of human leptospirosis. Overall accuracy of IgM ELISA was sensitivity of 87.9%, specificity of 97.0%, and area under the curve of 0...
April 2016: Diagnostic Microbiology and Infectious Disease
P B Shridhar, L W Noll, X Shi, B An, N Cernicchiaro, D G Renter, T G Nagaraja, J Bai
Shiga toxin-producing Escherichia coli (STEC) serogroups O26, O45, O103, O111, O121, and O145, called non-O157 STEC, are important foodborne pathogens. Cattle, a major reservoir, harbor the organisms in the hindgut and shed them in the feces. Although limited data exist on fecal shedding, concentrations of non-O157 STEC in feces have not been reported. The objectives of our study were (i) to develop and validate two multiplex quantitative PCR (mqPCR) assays, targeting O-antigen genes of O26, O103, and O111 (mqPCR-1) and O45, O121, and O145 (mqPCR-2); (ii) to utilize the two assays, together with a previously developed four-plex qPCR assay (mqPCR-3) targeting the O157 antigen and three virulence genes (stx1, stx2, and eae), to quantify seven serogroups and three virulence genes in cattle feces; and (iii) to compare the three mqPCR assays to a 10-plex conventional PCR (cPCR) targeting seven serogroups and three virulence genes and culture methods to detect seven E...
January 2016: Journal of Food Protection
Mariana Previato, Fábio Batista Frederico, Fernando Henrique Antunes Murata, Rubens Camargo Siqueira, Amanda Pires Barbosa, Aparecida Perpétuo Silveira-Carvalho, Cristina da Silva Meira, Vera Lúcia Pereira-Chioccola, Ricardo Gava, Plínio Pereira Martins Neto, Luiz Carlos de Mattos, Cinara Cássia Brandão de Mattos
BACKGROUND: Toxoplasmosis was recently included as a neglected disease by the Center for Disease Control. Ocular toxoplasmosis is one clinical presentation of congenital or acquired infection. The laboratory diagnosis is being used worldwide to support the clinical diagnosis and imaging. The aim of this study was to evaluate the use of serology and molecular methods to monitor acute OT in immunocompetent patients during treatment. METHODS: Five immunocompetent patients were clinically diagnosed with acute OT...
December 7, 2015: BMC Research Notes
Marcus Beissner, Richard Odame Phillips, Florian Battke, Malkin Bauer, Kossi Badziklou, Fred Stephen Sarfo, Issaka Maman, Agata Rhomberg, Ebekalisai Piten, Michael Frimpong, Kristina Lydia Huber, Dominik Symank, Moritz Jansson, Franz Xaver Wiedemann, Abiba Banla Kere, Karl-Heinz Herbinger, Thomas Löscher, Gisela Bretzel
BACKGROUND: As the major burden of Buruli ulcer disease (BUD) occurs in remote rural areas, development of point-of-care (POC) tests is considered a research priority to bring diagnostic services closer to the patients. Loop-mediated isothermal amplification (LAMP), a simple, robust and cost-effective technology, has been selected as a promising POC test candidate. Three BUD-specific LAMP assays are available to date, but various technical challenges still hamper decentralized application...
November 2015: PLoS Neglected Tropical Diseases
Lance W Noll, Pragathi B Shridhar, Xiaorong Shi, Baoyan An, Natalia Cernicchiaro, David G Renter, Tiruvoor G Nagaraja, Jianfa Bai
Several real-time polymerase chain reaction (PCR) assays have been developed to detect and quantify Shiga toxin-producing Escherichia coli (STEC) O157:H7, but none have targeted the O-antigen specific gene (rfbEO157) in combination with the three major virulence genes, stx1, stx2, and eae. Our objectives were to develop and validate a four-plex, quantitative PCR (mqPCR) assay targeting rfbE(O157), stx1, stx2, and eae for the detection and quantification of STEC O157 in cattle feces, and compare the applicability of the assay to detect STEC O157 to a culture method and conventional PCR (cPCR) targeting the same four genes...
September 2015: Foodborne Pathogens and Disease
Hamed Hasani, Mojtaba Bahrami, Abdorrasoul Malekpour, Mohammadreza Dehghani, Elaheh Allahyary, Mitra Amini, Mehdi Abdorahimi, Sara khani, Mohammad Kalantari Meibodi, Javad Kojuri
To determine the efficacy of different methods of cardiopulmonary resuscitation (CPCR) training in 3 different groups of the society. In a prospective and observational study of 2000 individuals in 3 different groups including G1, G2, and G3 4 different protocols of CPCR training were applied and their efficacy was compared between the groups. Also, 12 months after the study course, 460 participants from 3 groups were asked to take apart in a theoretical and practical examination to evaluate the long-term efficacy of the 4 protocols...
May 2015: Medicine (Baltimore)
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