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Molecular and Cellular Probes

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...
October 14, 2016: Molecular and Cellular Probes
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
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...
October 7, 2016: Molecular and Cellular Probes
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
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
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
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
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
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...
September 9, 2016: Molecular and Cellular Probes
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
Yang Yang, Xiaodong Qin, Wei Zhang, Yanmin Li, Zhidong Zhang
Porcine parvovirus (PPV) is a major cause of swine reproductive failure and reported in many countries worldwide. Recombinase polymerase amplification (RPA) assays using a real-time fluorescent detection (PPV real-time RPA assay) and a lateral flow dipstick (PPV RPA LFD assay) were developed targeting PPV NS1 gene. The detection limit of PPV real-time RPA assay was 300 copies per reaction within 9 min at 38 °C, while the RPA LFD assay has a detection limit of 400 copies per reaction in less than 20 min at 38 °C...
September 1, 2016: Molecular and Cellular Probes
Isfaqul Hussain, P Borah, R K Sharma, S Rajkhowa, M Rupnik, D P Saikia, D Hasin, Iftikar Hussain, N K Deka, L M Barkalita, Y Nishikawa, T Ramamurthy
A total of 1034 samples were collected from different sources and C. difficile was isolated from 18 (9.04%) of 199 human, 9 (4.89%) of 184 cattle, 29 (12.44%) of 233 pig, and from 23 (13.94%) of 165 poultry samples. Variations were observed on the rate of isolation according to age and clinical conditions (diarrhoea). None of the samples from cow, sheep, goat, local chicken, and wild animals yielded any C. difficile. Out of those isolates, 8, 2, 19 and 6 isolates from human, cattle, pig and poultry, respectively were toxigenic...
August 25, 2016: Molecular and Cellular Probes
Manfred Relle, Bernd Föhr, Federica Fasola, Andreas Schwarting
Granulomatosis with polyangiitis (GPA) is a severe autoimmune disease and one of the small vessel anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitides. Although its etiology and pathophysiology are still widely unknown, it is accepted that infections, environmental factors, epigenetic modifications, and a genetic predisposition provide the basis for this systemic disorder. GPA typically evolves into two phases: an initial phase characterized by ear, nose and throat (ENT) manifestations, such as chronic sinusitis and otitis, ulceration of the oral cavity and pharynx, as well as pulmonary nodules and a severe generalized phase, defined by the occurrence of rapidly progressive glomerulonephritis, pulmonary hemorrhage, and arthritis...
August 21, 2016: Molecular and Cellular Probes
Filipe Dantas-Torres, Kamila Gaudêncio da Silva Sales, Lidiane Gomes da Silva, Domenico Otranto, Luciana Aguiar Figueredo
We describe an improved real-time PCR assay for the detection and quantification of Leishmania infantum and Leishmania braziliensis kinetoplast DNA minicircles in canine blood samples. The analytical sensitivity of this technique is 0.1 fg of DNA per reaction, which is equivalent to 0.002 parasite. This assay uses a small reaction volume (15 μl) and is rapid to perform, with the results being available in less than 34 min. This improved assay might also be suitable for detecting and quantifying L. infantum and L...
August 20, 2016: Molecular and Cellular Probes
Vilija G Jokubaitis, Helmut Butzkueven
Multiple Sclerosis (MS) is an autoimmune degenerative disease of the central nervous system, characterized by multifocal demyelination and neurodegeneration. The genetic architecture of MS is complex, where genetic risk has been attributed to over 100 polymorphic loci each with small odds ratios. MS is a highly heterogeneous disease with numerous clinical and paraclinical endophenotypes. To-date, no genetic variant has been associated with clinical outcome, however, evidence exists that MS outcomes, like risk, are to an extent also controlled by genetic variation...
August 18, 2016: Molecular and Cellular Probes
Melanie Kauffmann, Steffen Rehbein, Dietmar Hamel, Walburga Lutz, Mike Heddergott, Kurt Pfister, Cornelia Silaghi
Infections with the tick-borne pathogens Anaplasma phagocytophilum and Babesia spp. can cause febrile disease in several mammalian species, including humans. Wild ruminants in Europe are suggested to serve as reservoir hosts for particular strains or species of these pathogens. The aims of this study were to investigate the occurrence of A. phagocytophilum and Babesia spp. in roe deer (Capreolus capreolus), fallow deer (Dama dama) and mouflon (Ovis musimon orientalis) in Germany, and the diversity and host association of genetic variants of A...
August 18, 2016: Molecular and Cellular Probes
Miljan Krstić, Simona Gabrielli, Marina Ignjatović, Sara Savić, Gabriella Cancrini, Gordana Ranđelović, Stefan Momčilović, Slavica Stojnev, Suzana Otašević
Dirofilaria immitis and D. repens are mosquito-borne nematodes that primarily infect canids, and can also cause mild to serious superficial or visceral infection in humans. In the present survey, peripheral blood from 150 asymptomatic dogs from Serbia were examined using the modified Knott's technique. Dirofilaria immitis, identified based on morphological and morphometric characteristics, was prevalent in dogs not receiving preventative treatment (in 44% and 60% of pound and pet dogs, respectively). These results, together with findings of autochthonous cases of subcutaneous D...
August 15, 2016: Molecular and Cellular Probes
Xing Ping Wang, Zhuo Ma Luoreng, Lin Sen Zan, Sayed Haidar Abbas Raza, Feng Li, Na Li, Shuan Liu
This study reports a significant up-regulation of bta-miR-146a and bta-miR-146b expression levels in bovine mammary tissues infected with subclinical, clinical and experimental mastitis. Potential target genes are involved in multiple immunological pathways. These results suggest a regulatory function of both miRNAs for the bovine inflammatory response in mammary tissue.
August 13, 2016: Molecular and Cellular Probes
Sally J Cutler, Eva Ruzic-Sabljic, Aleksandar Potkonjak
Lyme borreliosis (or Lyme disease) has become a virtual household term to the exclusion of other forgotten, emerging or re-emerging borreliae. We review current knowledge regarding these other borreliae, exploring their ecology, epidemiology and pathological potential, for example, for the newly described B. mayonii. These bacteria range from tick-borne, relapsing fever-inducing strains detected in some soft ticks, such as B. mvumii, to those from bat ticks resembling B. turicatae. Some of these emerging pathogens remain unnamed, such as the borrelial strains found in South African penguins and some African cattle ticks...
August 11, 2016: Molecular and Cellular Probes
Chien-Hsun Huang, Lina Huang, Mu-Tzu Chang, Kuo-Lung Chen
Members of the Bacillus subtilis group (BSG) possess industrial applicability; unfortunately, B. subtilis and its phylogenetically closest species are indistinguishable from one another using 16S rDNA sequencing, physiological and biochemical tests. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is a relatively novel technique for the fast and reliable identification of microorganisms. The aim of this study was to construct a unique analytical in-house database (IHDB) for BSG discrimination based on whole-cell protein fingerprinting using MALDI-TOF MS, as well as to discover biomarkers from the MS peaks to generate a classification model for further differentiation using the ClinProTools software...
August 6, 2016: Molecular and Cellular Probes
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