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Dna extraction

Layla Gomes Siravenha, Leonardo Quintão Siravenha, Lucimar Di Paula Madeira, Aldemir B Oliveira-Filho, Luiz Fernando Almeida Machado, Rosimar Neris Martins Feitosa, Antonio Carlos Rosário Vallinoto, Marluísa de Oliveira Guimarães Ishak, Ricardo Ishak
: The dental pulp is a sterile highly vascularized tissue and has been commonly used as a biological material to detect the genome of infectious agents that reach the dental tissue. Indeed, the pulp is also used to reveal past and ancient infections in the field of paleomicrobiology. The present study aimed to detect the presence of Hepatitis C virus (HCV) in a small community (approximately 400 inhabitants) in the Amazon region of Brazil (Nossa Senhora do Perpetuo Socorro, Vizeu, Para, Brazil) and standardize a technique for the detection of the virus in the dental pulp...
2016: PloS One
Thilini Piushani Keerthirathne, Dhammika Nayoma Magana-Arachchi, Dushantha Madegedara, Suneth Sithumini Sooriyapathirana
BACKGROUND: Mycobacteria have a spectrum of virulence and different susceptibilities to antibiotics. Distinguishing mycobacterial species is vital as patients with non-tuberculous mycobacterial (NTM) infections present clinical features that are similar to those of patients with tuberculosis. Thus, rapid differentiation of Mycobacterium tuberculosis complex from NTM is critical to administer appropriate treatment. Hence the aim of the study was to rapid identification of mycobacterial species present in bronchial washings using multiplex real time Polymerase Chain Reaction (PCR) and to determine the drug susceptibility in identified mycobacterial species...
October 26, 2016: BMC Infectious Diseases
Tereza Cervena, Andrea Rossnerova, Jitka Sikorova, Vit Beranek, Michal Vojtisek-Lom, Miroslav Ciganek, Jan Topinka, Pavel Rossner
Internal combustion engine emissions belong among the major anthropogenic sources of air pollution in urban areas. According to the International Agency for Research on Cancer (IARC), there is sufficient evidence of the carcinogenicity of diesel exhaust in humans. Although alternative fuels, mainly biodiesel, have recently become popular, little is still known about the genotoxicity of emissions from these fuels. We analysed DNA damage expressed as the frequency of micronuclei (MN) in human bronchial epithelial cells (BEAS-2B), induced by extractable organic matter (EOM; tested concentrations: 1, 10 and 25 μg/ml) obtained from particle emissions from various blends of biodiesel with diesel fuels (including neat diesel fuel (B0), a blend of 70% B0 and 30% biodiesel (B30) and neat biodiesel (B100))...
October 26, 2016: Basic & Clinical Pharmacology & Toxicology
Andrea Quagliariello, Irene Aloisio, Nicole Bozzi Cionci, Donata Luiselli, Giuseppe D'Auria, Llúcia Martinez-Priego, David Pérez-Villarroya, Tomaž Langerholc, Maša Primec, Dušanka Mičetić-Turk, Diana Di Gioia
Coeliac disease (CD) is associated with alterations of the intestinal microbiota. Although several Bifidobacterium strains showed anti-inflammatory activity and prevention of toxic gliadin peptides generation in vitro, few data are available on their efficacy when administered to CD subjects. This study evaluated the effect of administration for three months of a food supplement based on two Bifidobacterium breve strains (B632 and BR03) to restore the gut microbial balance in coeliac children on a gluten free diet (GFD)...
October 22, 2016: Nutrients
Alencar Kolinski Machado, Ana Cristina Andreazza, Tatiane Morgana da Silva, Aline Augusti Boligon, Vanusa do Nascimento, Gustavo Scola, Angela Duong, Francine Carla Cadoná, Euler Esteves Ribeiro, Ivana Beatrice Mânica da Cruz
Neuropsychiatric diseases, such as bipolar disorder (BD) and schizophrenia (SCZ), have a very complex pathophysiology. Several current studies describe an association between psychiatric illness and mitochondrial dysfunction and consequent cellular modifications, including lipid, protein, and DNA damage, caused by cellular oxidative stress. Euterpe oleracea (açaí) is a powerful antioxidant fruit. Açaí is an Amazonian palm fruit primarily found in the lowlands of the Amazonian rainforest, particularly in the floodplains of the Amazon River...
2016: Oxidative Medicine and Cellular Longevity
Jennyka Hallewell, Tim Reuter, Kim Stanford, Ed Topp, Trevor W Alexander
The goal of this study was to monitor Shiga toxin-producing Escherichia coli (STEC) serogroups and virulence genes in cattle (n = 30) originating from a closed herd. Fecal samples were collected (1) at weaning, (2) upon arrival to a feedlot, (3) after 30 days on feed (DOF), and (4) after 135 DOF. DNA was extracted from feces for detection of virulence and serogroup genes by polymerase chain reaction (PCR) and immunomagnetic separation and pulsed-field gel electrophoresis (PFGE) were performed to collect and subtype STEC isolates...
October 25, 2016: Foodborne Pathogens and Disease
Virginie Gaget, Angela Keulen, Melody Lau, Paul Monis, Justin D Brookes
AIMS: Benthic cyanobacteria produce toxic and odorous compounds similar to their planktonic counterparts, challenging the quality of drinking water supplies. The biofilm that benthic algae and other micro-organisms produce is a complex and protective matrix. Monitoring to determine the abundance and identification of cyanobacteria therefore relies on molecular techniques, with the choice of DNA isolation technique critical. This study investigated which DNA extraction method is optimal for DNA recovery in order to guarantee the best DNA yield for PCR-based analysis of benthic cyanobacteria...
October 25, 2016: Journal of Applied Microbiology
Wei Lu, Jia-Lin Yu, Lu-Quan Li, Qi Lu, Zheng-Li Wang, Yun Pan
Bacterial infectious pneumonia is one of the major causes of mortality in neonates, particularly when the neonates suffer from ventilator‑associated pneumonia (VAP). However, the causes of pneumonia are difficult to define. Thus, the present study focused on understanding the diversity of microflora in the lower respiratory tract to elucidate the causes. The experimental groups comprised newborns who suffered from infectious pneumonia with or without VAP (IVAP and IP groups, respectively), whereas the control group comprised newborns who suffered from respiratory distress syndrome (RDS) without VAP (RDS group)...
October 25, 2016: Molecular Medicine Reports
Han-Shi Zeng, Wei-Xia Lin, Shu-Tao Zhao, Zhan-Hui Zhang, Heng-Wen Yang, Feng-Ping Chen, Yuan-Zong Song, Zhi-Nan Yin
Neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) is an autosomal recessive disorder resulting from biallelic mutations of the SLC25A13 gene. Due to the lack of well‑recognized clinical or biochemical diagnostic criteria, the definitive diagnosis of this disease relies on the genetic analysis of SLC25A13 at present. As novel large deletion/insertion mutations of the SLC25A13 gene are difficult to detect using routine DNA analytic approaches, the timely diagnosis of patients with these types of mutations remains a challenge...
October 21, 2016: Molecular Medicine Reports
D-X Ban, Y Liu, T-W Cao, S-J Gao, S-Q Feng
STUDY DESIGN: The rat's acellular spinal cord scaffold (ASCS) and spinal cord neurons were prepared in vitro to explore their biocompatibility. OBJECTIVES: The preparation of ASCS and co-culture with neuron may lay a foundation for clinical treatment of spinal cord injury (SCI). SETTING: Tianjin Medical University General Hospital, ChinaMethods:ASCS was prepared by chemical extraction method. Hematoxylin and eosin (H&E), myelin staining and scanning electron microscope were used to observe the surface structure of ASCS...
October 25, 2016: Spinal Cord
María Belén Cerliani, Walter Pavicic, Juan Antonio Gili, Graciela Klein, Silvia Saba, Silvina Richard
AIM: To analyze the association between oncohematological diseases and GSTT1/GSTM1/CYP1A1 polymorphisms, dietary habits and smoking, in an argentine hospital-based case-control study. METHODS: This hospital-based case-control study involved 125 patients with oncohematological diseases and 310 control subjects. A questionnaire was used to obtain sociodemographic data and information about habits. Blood samples were collected, and DNA was extracted using salting out methods...
October 10, 2016: World Journal of Clinical Oncology
Shuaihua Pu, Hamidreza Khazanehei, Peter J Jones, Ehsan Khafipour
Long-term dietary fatty acid intake is believed to induce changes in the human gut microbiome which might be associated with human health or obesity status; however, considerable debate remains regarding the most favorable ratios of fatty acids to optimize these processes. The objective of this sub-study of a double-blinded randomized crossover clinical study, the canola oil multi-center intervention trial, was to investigate effects of five different novel oil blends fed for 30 days each on the intestinal microbiota in 25 volunteers with risk of metabolic syndrome...
2016: Frontiers in Microbiology
Xiao-Yu Hu, Yu-Fei Yao, Bo-Miao Cui, Jun Lv, Xin Shen, Biao Ren, Ming-Yun Li, Qiang Guo, Rui-Jie Huang, Yan Li
OBJECTIVE: To analyze the whole microbial structure in a case of rampant caries to provide evidence for its prevention and treatment. METHODS: Clinical samples including blood, supragingival plaque, plaque in the caries cavity, saliva, and mucosal swabs were collected with the patient's consent. The blood sample was sent for routine immune test, and the others samples were stained using Gram method and cultured for identifying colonies and 16S rRNA sequencing. DNA was extracted from the samples and tested for the main cariogenic bacterium (Streptococcus mutans) with qPCR, and the whole microbial structure was analyzed using DGGE...
October 20, 2016: Nan Fang Yi Ke da Xue Xue Bao, Journal of Southern Medical University
Rangel Moreira Silva, Luciane Dias Pereira, Jefferson Hollanda Véras, Camila Regina do Vale, Lee Chen-Chen, Suzana da Costa Santos
Ellagitannins are well-known antioxidants in medicinal plants, foods, and edible fruits, particularly in Myrciaria cauliflora (jabuticaba). Thus, this study aimed to evaluate the protective effects of jabuticaba seed extract (JSE) and pedunculagin using in vivo micronucleus test and comet assay in mouse bone marrow cells, in combination with cyclophosphamide (CP), a bioreductive alkylating agent. The ellagitannin composition of JSE was determined by HPLC/PDA, with castalagin, vescalagin, and pedunculagin as the main compounds (124...
November 1, 2016: Mutation Research
Hala Attia, Nouf Al-Rasheed, Raeesa Mohamad, Nawal Al-Rasheed, Maha Al-Amin
BACKGROUND: Hepatic fibrosis and its end point; cirrhosis, are the major cause of liver failure and death in patients with chronic liver disease. Therefore, the need for an effective treatment is evident. This study was designed to assess the potential effects of aqueous extract of date fruits, either flesh (DFE) or pits (DPE), on oxidative DNA damage and liver inflammation induced by carbon tetrachloride (CCl4) and whether they are related to inhibition of nuclear factor-κB pathway...
October 24, 2016: BMC Complementary and Alternative Medicine
Tahira Younis, Muhammad Rashid Khan, Moniba Sajid
BACKGROUND: Leaves and root bark of Fraxinus xanthoxyloides Wall. (Oleaceae) are used locally for the treatment of jaundice, malaria and pneumonia. Decoction of stem, twigs and bark is used in pain, internal injuries, rheumatism and in bone fracture. In this investigation we have evaluated the methanol extract of leaves for its hepatoprotective potential against CCl4 induced hepatic injuries in rat. METHODS: Powder of F. xanthoxyloides leaves was extracted with methanol (FXM) and subjected for the determination of polyphenolics through HPLC-DAD analysis...
October 24, 2016: BMC Complementary and Alternative Medicine
Wipada Duangprompo, Kalaya Aree, Arunporn Itharat, Pintusorn Hansakul
5,6-dihydroxy-2,4-dimethoxy-9,10-dihydrophenanthrene (HMP) is an active compound isolated from the rhizome extracts of Dioscorea membranacea Pierre, a Thai medicinal plant. This study aimed to investigate the growth-inhibitory and apoptosis-inducing effects of HMP in human lung cancer A549 cells. The antiproliferative and cytotoxic effects of HMP were analyzed by a Sulforhodamine B assay. Cell division, cell cycle distribution and membrane asymmetry changes were each performed with different fluorescent dyes and then analyzed by flow cytometry...
October 25, 2016: American Journal of Chinese Medicine
Kimiko Aoki, Hiroyuki Tanaka, Makoto Ueki
When the tampering of a urine sample is suspected in doping control, personal identification of the sample needs to be determined by short tandem repeat (STR) analysis using DNA. We established a method for extracting DNA from urine samples stored at -20 °C without using any additives or procedures, which is consistent with how samples are required to be managed for doping control. The method, using the Puregene® Blood Core kit followed by NucleoSpin® gDNA Clean-up or NucleoSpin® gDNA Clean-up XS kit, does not need any special instrument and can provide a purified extract with high-quality DNA from up to 40 mL of urine suitable for STR analysis using an AmpFlSTR® Identifiler® PCR amplification kit...
October 24, 2016: Drug Testing and Analysis
Alfred Chikweto, Ravindra N Sharma, Keshaw P Tiwari, Shiv K Verma, Rafael Calero-Bernal, Tiantian Jiang, Chunlei Su, Oliver C Kwok, Jitender P Dubey
The objectives of the present cross sectional study were to isolate and genotype Toxoplasma gondii in free-range chickens from Grenada, West Indies. Using the modified agglutination test, antibodies to T. gondii were found in 39 (26.9%) of 145 free-range chickens with titers of 25 in 7chickens; 50 in 6 chickens; 100 in 2 chickens; 200 or higher in 24 chickens. The hearts of the 39 seropositive chickens were bioassayed in mice; viable T. gondii was isolated from 20 and further propagated in cell culture. Genotyping of T...
October 24, 2016: Journal of Parasitology
Zhenqin Wu, Huimin Bi, Sichen Pan, Lingyi Meng, Xin Sheng Zhao
Fluorescence correlation spectroscopy (FCS) is a powerful tool to investigate molecular diffusion and relaxations, which may be utilized to study many problems, such as molecular size and aggregation, chemical reaction, molecular transportation and motion, and various kinds of physical and chemical relaxations. This paper focuses on a problem related to using the relaxation term to study a reaction. If two species with different fluorescence photon emission efficiencies are connected by a reaction, the kinetic and equilibrium properties will be manifested in the relaxation term of the FCS curve...
October 24, 2016: Journal of Physical Chemistry. B
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