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Molecular and Cellular Probes

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https://www.readbyqxmd.com/read/27894847/pcr-screening-of-tick-borne-agents-in-sensitive-conservation-areas-southeast-portugal
#1
Maria Margarida Santos-Silva, Pedro Melo, Nuno Santos, Sandra Antunes, Luís Raposo Duarte, Joana Ferrolho, Natacha Milhano, Patrícia Tavares Santos, Ana Domingos, Ana Sofia Santos
The Southeast region of Portugal, particularly the Guadiana valley, is currently the reintroduction territory of Lynx pardinus (Iberian lynx), one of the most endangered felids in the world that is only found in the Iberian Peninsula. Over the last century, populations have declined, placing L. pardinus at extremely high risk of extinction in the wild and relying on reintroduction projects. Among the aspects taken into account in the establishment of new populations is the sanitary status of the selected habitats, especially concerning infectious diseases, including tick-borne pathogens (TBPs)...
November 25, 2016: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/27890610/a-novel-real-time-pcr-assay-for-highly-specific-detection-and-quantification-of-vaginal-lactobacilli
#2
Vladimir V Demkin, Stanislav I Koshechkin, Alexei Slesarev
PCR detection and quantification of vaginal lactobacilli remains problematic because of the high level of genetic heterogeneity and taxonomic complexity within the genus Lactobacillus. The aim of the present study was to identify conserved sequences among the genomes of major species of vaginal lactobacilli that could be used for the development of a PCR-based method for quantitative determination of vaginal microbiota-specific lactobacilli. Comparative analysis of the genomes of several species of vaginal lactobacilli allowed us to identify conserved regions in the rplK gene, which encodes ribosomal protein L11, and to design group-specific PCR primers and a probe for selected species from the L...
November 24, 2016: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/27871797/direct-pcr-a-rapid-method-for-multiplexed-detection-of-different-serotypes-of-salmonella-in-enriched-pork-meat-samples
#3
Wai Hoe Chin, Yi Sun, Jonas Høgberg, Than Linh Quyen, Pia Engelsmann, Anders Wolff, Dang Duong Bang
Salmonellosis, an infectious disease caused by Salmonella spp., is one of the most common foodborne diseases. Isolation and identification of Salmonella by conventional bacterial culture method is time consuming. In response to the demand for rapid on line or at site detection of pathogens, in this study, we developed a multiplex Direct PCR method for rapid detection of different Salmonella serotypes directly from pork meat samples without any DNA purification steps. An inhibitor-resistant Phusion Pfu DNA polymerase was used to overcome PCR inhibition...
November 18, 2016: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/27856333/novel-homozygous-missense-mutation-in-aldh7a1-causes-neonatal-pyridoxine-dependent-epilepsy
#4
Emanuele G Coci, Luca Codutti, Christian Fink, Sophie Bartsch, Gunnar Grüning, Thomas Lücke, Ingo Kurth, Joachim Riedel
Pyridoxine dependent epilepsy (PDE) (OMIM#266100) is a neonatal form of epilepsy, caused by dysfunction of the enzyme α-aminoadipic semialdehyde dehydrogenase (ALDH7A1 or Antiquitin). This enzyme converts α-aminoadipic semialdehyde (α-AASA) into α-aminoadipate (AAA), a critical step in the lysine metabolism of the brain. ALDH7A1 dysfunction causes an accumulation of α-AASA and δ(1)-piperideine-6-carboxylic acid (P6C), which are in equilibrium with each other. P6C binds and inactivates pyridoxal 5'-phosphate (PLP), the active form of pyridoxine...
November 14, 2016: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/27777104/west-nile-virus-circulation-in-vojvodina-serbia-mosquito-bird-horse-and-human-surveillance
#5
Dušan Petrić, Tamaš Petrović, Ivana Hrnjaković Cvjetković, Marija Zgomba, Vesna Milošević, Gospava Lazić, Aleksandra Ignjatović Ćupina, Diana Lupulović, Sava Lazić, Dragan Dondur, Slavica Vaselek, Aleksandar Živulj, Bratislav Kisin, Tibor Molnar, Djordje Janku, Dubravka Pudar, Jelena Radovanov, Mihaela Kavran, Gordana Kovačević, Budimir Plavšić, Aleksandra Jovanović Galović, Milan Vidić, Svetlana Ilić, Mina Petrić
Efforts to detect West Nile virus (WNV) in the Vojvodina province, northern Serbia, commenced with human and mosquito surveillance in 2005, followed by horse (2009) and wild bird (2012) surveillance. The knowledge obtained regarding WNV circulation, combined with the need for timely detection of virus activity and risk assessment resulted in the implementation of a national surveillance programme integrating mosquito, horse and bird surveillance in 2014. From 2013, the system showed highly satisfactory results in terms of area specificity (the capacity to indicate the spatial distribution of the risk for human cases of West Nile neuroinvasive disease - WNND) and sensitivity to detect virus circulation even at the enzootic level...
October 21, 2016: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/27771442/serum-based-fluorescent-assay-for-evaluating-dipeptidyl-peptidase-i-activity-in-collagen-induced-arthritis-rat-model
#6
Xiaoqian Liu, Jingjing Wang, Yi Chu, Xiaoying Zhou
Dipeptidyl peptidase I (DPPI) is a lysosomal cysteine protease and derived from immune granule cells. It has been suggested playing an important role in the development of rheumatoid arthritis. In this study, a coumarin based fluorescent probe (GF-AFC) was designed and synthesized to evaluate DPPI activity in serum or tissue homogenates of collagen-induced arthritis (CIA) rats, an inflammatory arthropathy model. It was revealed that the fluorescent intensity was significantly increased in a very short time after specific substrate GF-AFC reacted with the DPPI...
October 19, 2016: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/27771441/label-free-monitoring-of-dna-polymerase-activity-based-on-a-thrombin-binding-aptamer-g-quadruplex
#7
Jing Wang, Haisheng Liu, Changbei Ma, Jun Wang, Linxiu Zhong, Kefeng Wu
We have developed a label-free assay for the detection of DNA polymerase activity based on a thrombin-binding aptamer (TBA) G-quadruplex. In the presence of DNA polymerase, the 3'-OH termini of the hairpin substrate are immediately elongated to replace the TBA, which can be recognized quickly by the ThT dye and results in an increase of fluorescence. This method is highly sensitive with a detection limit of 0.1 U/mL. It is simple and cost-effective without any requirement of labeling with a fluorophore-quencher pair...
October 19, 2016: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/27765650/a-pcr-based-protocol-to-accurately-size-c9orf72-intermediate-length-alleles
#8
Giorgio Biasiotto, Silvana Archetti, Diego Di Lorenzo, Francesca Merola, Giulia Paiardi, Barbara Borroni, Antonella Alberici, Alessandro Padovani, Massimiliano Filosto, Cristian Bonvicini, Luigi Caimi, Isabella Zanella
Although large expansions of the non-coding GGGGCC repeat in C9orf72 gene are clearly defined as pathogenic for Amyotrophic Lateral Sclerosis (ALS) and Frontotemporal Lobar Degeneration (FTLD), intermediate-length expansions have also been associated with those and other neurodegenerative diseases. Intermediate-length allele sizing is complicated by intrinsic properties of current PCR-based methodologies, in that somatic mosaicism could be suspected. We designed a protocol that allows the exact sizing of intermediate-length alleles, as well as the identification of large expansions...
October 17, 2016: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/27746242/development-of-a-real-time-pcr-for-bartonella-spp-detection-a-current-emerging-microorganism
#9
Elena Parra, Ferran Segura, Jessica Tijero, Imma Pons, Maria-Mercedes Nogueras
A real-time PCR assay using SYBR Green was optimized to detect those Bartonella that are most frequently described as pathogens. The assay was genus-specific. Sequencing allowed to distinguish species. Assay sensitivity was determined using 10-fold serial dilutions of genomic DNA. Dynamic range was 100 ng-100 fg and sensitivity was 50 copies/reaction.
October 13, 2016: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/27725294/aberrant-expression-of-mir-9-in-benign-and-malignant-breast-tumors
#10
Aisan Hasanzadeh, Hamzeh Mesrian Tanha, Kamran Ghaedi, Mahboobeh Madani
PURPOSE: MicroRNAs (miRNAs) are involved in the progression of breast cancer (BC). miR-9 has been reported to be correlated with either favorable or unfavorable events in BC. This study was aimed to evaluate the expression level of miR-9 in human breast tissues, including benign and malignant tumor samples and also healthy tissue. MATERIALS AND METHODS: The expression level of miR-9 was analyzed in 10 normal breast tissues, 30 malignant, and 30 benign breast tumor tissue samples using RT-PCR and qPCR...
October 7, 2016: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/27720908/molecular-detection-of-mycoplasma-haemomuris-subspecies-using-dnak-targeted-real-time-pcr-with-sybr-green-i-and-melting-curve-analysis
#11
Hiroshi Satoh, Daiki Imai, Shigeru Sato, Kazuhisa Furuhama, Ryô Harasawa
Hemoplasmas cause severe infections in mammals, but these pathogens are difficult to detect and identify at the species and subspecies level because of the need for time-consuming sequence based methods. Here, we used real-time PCR with SYBR Green I targeting of the dnaK gene followed by standard melting curve analysis to achieve rapid detection and differentiation of the Mycoplasma haemomuris subspecies 'Candidatus Mycoplasma haemomuris subsp. musculi' and 'Candidatus M. haemomuris subsp. ratti'. The melting temperatures of the PCR products, 84...
October 6, 2016: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/27720907/molecular-beacons-with-joe-dye-influence-of-linker-and-3-couple-quencher
#12
Dmitry A Tsybulsky, Maksim V Kvach, Dmitry Y Ryazantsev, Ilya O Aparin, Alexander A Stakheev, Igor A Prokhorenko, Yury V Martynenko, Sergey V Gontarev, Andrey A Formanovsky, Timofei S Zatsepin, Vadim V Shmanai, Vladimir A Korshun, Sergey K Zavriev
Molecular beacons carrying JOE dye (4',5'-dichloro-2',7'-dimethoxy-6-carboxyfluorescein) on a rigid or flexible linker and one or two BHQ1 quenchers have been prepared and tested in real-time PCR using Fusarium avenaceum elongation factor 1α DNA template. The probes were different in their structures (loop size and stem length), linkers for dye attachment (6-aminohexanol or trans-4-aminocyclohexanol), quencher composition (single and double BHQ1) to elucidate the influence of all these features. Fluorogenic properties of the probes were studied and compared to those of FAM (fluorescein)-based probes...
October 6, 2016: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/27720768/application-of-immuno-pcr-assay-for-the-detection-of-serum-ige-specific-to-bermuda-allergen
#13
Samina Rahmatpour, Amjad Hayat Khan, Rasoul Nasiri Kalmarzi, Masoumeh Rajabibazl, Gholamreza Tavoosidana, Elahe Motevaseli, Nosratollah Zarghami, Esmaeil Sadroddiny
In vivo and in vitro tests are the two major ways of identifying the triggering allergens in sensitized individuals with allergic symptoms. Both methods are equally significant in terms of sensitivity and specificity. However, in certain circumstances, in vitro methods are highly preferred because they circumvent the use of sensitizing drugs in patients. In current study, we described a highly sensitive immuno-PCR (iPCR) assay for serum IgE specific to Bermuda allergens. Using oligonucleotide-labelled antibody, we used iPCR for the sensitive detection of serum IgE...
October 5, 2016: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/27687066/four-novel-polymorphisms-of-buffalo-insig2-gene-are-associated-with-milk-production-traits-in-chinese-buffaloes
#14
Tingxian Deng, Chunying Pang, Xiaoya Ma, Xingrong Lu, Anqin Duan, Peng Zhu, Xianwei Liang
Insulin-induced genes (INSIGs), including INSIG1 and INSIG2, are important mediators that play a pivotal role in the lipid metabolism and could cause the retention of the SCAP/SREBP complex. Therefore, the objective of this study is to detect the single nucleotide polymorphisms (SNPs) of buffalo INSIG2 gene and evaluate their associations with milk production traits in Chinese buffaloes. A total of four SNPs (g.621272A > G, g.621364A > C, g.632543G > A, and g.632684C > T) were identified using DNA pooled sequencing, and the SNP genotyping for the identified SNPs was performed by using Matrix-assisted laser desorption/ionization time of flight mass spectrometry method from 264 individuals...
September 27, 2016: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/27618775/prediction-of-post-translation-modifications-at-the-contact-site-between-anaplasma-phagocytophilum-and-human-host-during-autophagosome-induction-using-a-bioinformatic-approach
#15
Zarrin Basharat, Sarah Rizwan Qazi, Azra Yasmin, Syed Aoun Ali, Deeba Noreen Baig
Autophagy is crucial for maintaining physiological homeostasis, but its role in infectious diseases is not yet adequately understood. The binding of Anaplasma translocated substrate-1 (ATS1) to the human Beclin1 (BECN1) protein is responsible for the modulation of autophagy pathway. ATS1-BECN1 is a novel type of interaction that facilitates Anaplasma phagocytophilum proliferation, leading to intracellular infection via autophagosome induction and segregation from the lysosome. Currently, there is no report of post translational modifications (PTMs) of BECN1 or cross-talk required for ATS-BECN1 complex formation...
September 9, 2016: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/27888871/dissecting-the-genetic-background-of-multifactorial-diseases-and-traits-a-major-challenge-for-genetic-research
#16
EDITORIAL
Sabine Hoffjan
No abstract text is available yet for this article.
December 2016: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/27818248/genetics-of-parkinson-s-disease
#17
REVIEW
Christina M Lill
Almost two decades after the identification of SNCA as the first causative gene in Parkinson's disease (PD) and the subsequent understanding that genetic factors play a substantial role in PD development, our knowledge of the genetic architecture underlying this disease has vastly improved. Approximately 5-10% of patients suffer from a monogenic form of PD where autosomal dominant mutations in SNCA, LRRK2, and VPS35 and autosomal recessive mutations in PINK1, DJ-1, and Parkin cause the disease with high penetrance...
December 2016: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/27751841/if-genetic-variation-could-talk-what-genomic-data-may-teach-us-about-the-importance-of-gene-expression-regulation-in-the-genetics-of-autism
#18
REVIEW
Erika Yeh, Lauren A Weiss
Autism spectrum disorder (ASD) has been long known to have substantial genetic etiology. Much research has attempted to identify specific genes contributing to ASD risk with the goal of tying gene function to a molecular pathological explanation for ASD. A unifying molecular pathology would potentially increase understanding of what is going wrong during development, and could lead to diagnostic biomarkers or targeted preventative or therapeutic directions. We review past and current genetic mapping approaches and discuss major results, leading to the hypothesis that global dysregulation of gene or protein expression may be implicated in ASD rather than disturbance of brain-specific functions...
December 2016: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/27725295/trying-to-understand-the-genetics-of-atopic-dermatitis
#19
REVIEW
Susanne Stemmler, Sabine Hoffjan
Atopic dermatitis (AD) is a common and complex skin disease associated with both genetic and environmental factors. Loss-of-function mutations in the filaggrin gene, encoding a structural protein with an important role in epidermal barrier function, constitutes a well recognised susceptibility locus for AD. Further, genome-wide association studies (GWAS), including large meta-analyses, have discovered 38 additional susceptibility loci with genome-wide significance. However, the reported variations only explain a fraction of the overall heritability of AD...
December 2016: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/27618776/genomics-of-alzheimer-s-disease-value-of-high-throughput-genomic-technologies-to-dissect-its-etiology
#20
REVIEW
Giuseppe Tosto, Christiane Reitz
Late-onset Alzheimer's disease (AD), the most common neurodegenerative disorder in western countries, is clinically defined by progressive worsening in cognitive functions along with function and behavioral impairment. This ultimately results in complete incapacity and death. AD is a clinically and pathologically heterogeneous disease, and this is reflected by the numerous genetic findings that point to several diverse molecular mechanisms and pathways. Linkage, genome-wide association and next-generation sequencing studies have led to the identification of more than 20 novel susceptibility loci for AD...
December 2016: Molecular and Cellular Probes
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