Isolation of Tn916-like conjugal elements from swine lot effluent

2000 ◽  
Vol 46 (6) ◽  
pp. 542-549 ◽  
Author(s):  
Bradley J Haack ◽  
Robert E Andrews Jr.
Keyword(s):  
Bacillus Thuringiensis ◽  
Enterococcus Faecalis ◽  
Tetracycline Resistance ◽  
Bacillus Species ◽  
Pcr Analysis ◽  
Soil Environment ◽  
Swine Waste ◽  
Chain Reaction ◽  
Plasmid Pc194 ◽  
Polymerase Chain

Isolates of Enterococcus faecalis obtained from a swine farrowing house outflow were examined for genetic elements similar to Tn916. Of the enterococci isolated, 71% were resistant to tetracycline. Among the tetracycline-resistant enterococci isolated from the outflow samples, approximately 34% were able to transfer the tetracycline resistance phenotype to Bacillus thuringiensis in cross-genus matings. The frequencies of transfer for 10 random isolates were comparable to those for transfer of Tn916 from E. faecalis to B. thuringiensis. In addition, these elements were shown to mobilize plasmid pC194 between Bacillus species, as did Tn916. Southern blot and polymerase chain reaction (PCR) analysis showed these elements share extensive structural homology with Tn916. The selected conjugal elements were capable of transfer to a Bacillus recipient in a soil environment. When the swine waste was introduced into the soil, the tetracycline resistant fecal enterococci levels rose from essentially undetectable levels to approximately 4 × 104and remained at this level for 4 weeks. After six months, including one winter, levels had decreased to 5 × 103.Key words: Enterococcus faecalis, Tn916, swine waste, genetic exchange, Bacillus thuringiensis, PCR.

Sensitivity of polymerase chain reaction (PCR)-southern hybridization and conventional PCR analysis for Halal authentication of gelatin capsules

LWT ◽  
2015 ◽  
Vol 63 (1) ◽  
pp. 714-719 ◽  
Author(s):  
Sahilah Abd Mutalib ◽  
Nursheila Mustafa Muin ◽  
Aminah Abdullah ◽  
Osman Hassan ◽  
Wan Aida Wan Mustapha ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Southern Hybridization ◽  
Pcr Analysis ◽  
Conventional Pcr ◽  
Chain Reaction ◽  
Gelatin Capsules ◽  
Halal Authentication ◽  
Polymerase Chain

Development of Three-Stage Bioaerosol Sampler for Size-Selective Sampling

2022 ◽  
Author(s):  
Jun-Hyung Lim ◽  
Sang Hwan Nam ◽  
Jongwoo Kim ◽  
Nam Hoon Kim ◽  
Gun-Soo Park ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Size Range ◽  
Collection Efficiency ◽  
Commercial Product ◽  
Pcr Analysis ◽  
Cyclone Separator ◽  
Chain Reaction ◽  
Selective Sampling ◽  
Sampling Flow Rate ◽  
Polymerase Chain

Abstract In this study, a three-stage bioaerosol sampler with a sampling flow rate of 170 L/min was designed and fabricated for sampling the bioaerosols released during human breathing and coughing, and its performance was evaluated. The sampler was constructed using a cyclone separator with a cutoff size of 2.5 µm as a preseparator, a multi-nozzle virtual impactor with a cutoff size of 0.34 µm as an aerosol concentrator, and a BioSampler, which is a commercial product, for collecting bioaerosols in a collection fluid. The collection efficiency of the sampler was evaluated through simulations and experiments. Only particles with sizes of 0.1-4 µm were selectively collected in the collection fluid. Bacteriophage bioaerosols were sampled using the developed sampler and ACD-200 Bobcat sampler, which is a commercial product. The amounts of collected bacteriophages were compared using the polymerase chain reaction (PCR) technique. The sampling performance of the developed sampler was similar to that of the ACD-200 Bobcat sampler. Moreover, the developed sampler showed its ability to sample bioaerosols of a specific size-range and collect them directly in a collection fluid for the PCR analysis. Therefore, the developed sampler is expected to be useful for indoor environmental monitoring by effectively sampling the bioaerosols released indoors during human breathing and coughing.

scholarly journals Aberrant Long Noncoding RNAs Expression Profiles Affect Cisplatin Resistance in Lung Adenocarcinoma

2017 ◽  
Vol 2017 ◽  
pp. 1-14 ◽  
Author(s):  
Lijuan Hu ◽  
Jian Chen ◽  
Fan Zhang ◽  
Junjun Wang ◽  
Jingye Pan ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
A549 Cell ◽  
Cisplatin Resistance ◽  
Expression Profiles ◽  
Noncoding Rnas ◽  
Long Noncoding Rnas ◽  
Pcr Analysis ◽  
Sensitive Cell ◽  
Chain Reaction ◽  
Polymerase Chain

Background. Long noncoding RNAs (lncRNAs) have been shown to be involved in the mechanism of cisplatin resistance in lung adenocarcinoma (LAD). However, the roles of lncRNAs in cisplatin resistance in LAD are not well understood. Methods. We used a high-throughput microarray to compare the lncRNA and mRNA expression profiles in cisplatin resistance cell A549/DDP and cisplatin sensitive cell A549. Several candidate cisplatin resistance-associated lncRNAs were verified by real-time quantitative reverse transcription polymerase chain reaction (PCR) analysis. Results. We found that 1,543 lncRNAs and 1,713 mRNAs were differentially expressed in A549/DDP cell and A549 cell, hinting that many lncRNAs were irregular from cisplatin resistance in LAD. We also obtain the fact that 12 lncRNAs were aberrantly expressed in A549/DDP cell compared with A549 cell by quantitative PCR. Among these, UCA1 was the aberrantly expressed lncRNA and can significantly reduce the IC50 of cisplatin in A549/DDP cell after knockdown, while it can increase the IC50 of cisplatin after UCA1 was overexpressed in NCI-H1299. Conclusions. We obtained patterns of irregular lncRNAs and they may play a key role in cisplatin resistance of LAD.

scholarly journals Suitability of different tissue fixatives for subsequent PCR analysis of Cysticercus ovis

2012 ◽  
Vol 49 (2) ◽  
pp. 67-70 ◽  
Author(s):  
M. Kolesárová ◽  
R. Herich ◽  
M. Levkut ◽  
J. Čurlík ◽  
M. Levkut
Keyword(s):  
Retrospective Studies ◽  
Pcr Amplification ◽  
Pcr Analysis ◽  
Chain Reaction ◽  
Fresh Tissue ◽  
Carnoy’S Solution ◽  
Negative Results ◽  
Pcr Product ◽  
Polymerase Chain ◽  
Cysticercus Ovis

AbstractPCR amplification of specific DNA regions is a powerful tool for retrospective studies, but not all preservation or fixation methods render DNA that is suitable for subsequent amplification. Several factors affect sensitivity of polymerase chain reaction (PCR) amplification. There were reported the effects of commonly used fixation solutions — 10 % neutral buffered formalin, 20 % neutral buffered formalin and Carnoy’s solution and the efficiency of PCR amplification in fresh tissue and paraffin (or wax) embedded samples of Cysticercus ovis. DNA from samples was isolated and PCR product of 1300 bp was amplified. Results indicated that the samples fixed in Carnoy’s solution produced reliable amplification of desired fragments. The samples that were fixed in 10 % and 20 % neutral buffered formalin brought negative results.

Quantitative PCR Analysis for Bcl-2/IgH in a Phase III Study of Yttrium-90 Ibritumomab Tiuxetan As Consolidation of First Remission in Patients With Follicular Lymphoma

2009 ◽  
Vol 27 (36) ◽  
pp. 6094-6100 ◽  
Author(s):  
Lindsey Goff ◽  
Karin Summers ◽  
Sameena Iqbal ◽  
Jens Kuhlmann ◽  
Michael Kunz ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Follicular Lymphoma ◽  
Quantitative Polymerase Chain Reaction ◽  
Phase Iii ◽  
Pcr Analysis ◽  
Control Group ◽  
Ibritumomab Tiuxetan ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Yttrium 90

Purpose The randomized First-Line Indolent Trial (FIT) was conducted in patients with advanced follicular lymphoma (FL), to evaluate the safety and efficacy of yttrium-90 (90Y) ibritumomab tiuxetan given as consolidation of complete or partial remission. This study of minimal residual disease was undertaken in parallel, to determine the rate of conversion from bcl-2 polymerase chain reaction (PCR) –detectable to –undetectable status and the corresponding effect on progression-free survival (PFS). Patients and Methods Blood samples from 414 patients (90Y-ibritumomab, n = 208; control, n = 206) were evaluated using real-time quantitative polymerase chain reaction (RQ-PCR); 186 were found to have the bcl-2 rearrangement and were thus eligible for inclusion in the RQ-PCR analysis. Results Overall, 90% of treated patients converted from bcl-2 PCR–detectable to –undetectable disease status, compared with 36% in the control group. Treatment significantly prolonged median PFS in patients converting to bcl-2 PCR-undetectable status (40.8 v 24.0 months in the control group; P < .01, hazard ratio [HR], 0.399). In patients who had bcl-2 PCR-detectable disease at random assignment, treatment significantly prolonged median PFS (38.4 v 8.2 months in the control group; P < .01, HR, 0.293). Conclusion Eradication of PCR-detectable disease occurred more frequently after treatment with 90Y-ibritumomab tiuxetan and was associated with prolongation of PFS.

scholarly journals Xuezhikang Therapy Increases miR-33 Expression in Patients with Low HDL-C Levels

2014 ◽  
Vol 2014 ◽  
pp. 1-5 ◽  
Author(s):  
Ruihua Cao ◽  
Yongyi Bai ◽  
Lan Sun ◽  
Jin Zheng ◽  
Mian Zu ◽  
...  
Keyword(s):  
Hdl Cholesterol ◽  
Pcr Analysis ◽  
Chain Reaction ◽  
Low Hdl ◽  
The Mean ◽  
Polymerase Chain ◽  
Baseline Levels ◽  
Hdl Metabolism ◽  
To Receive ◽  
Plasma Mirnas

Background. MicroRNA-33a and -b (miR-33a/b) have been revealed to be posttranscriptional regulators of HDL metabolism. Xuezhikang (XZK) is a marked natural HDL-raising polypill. We aim to evaluate the effects of XZK on the expression of circulating miR-33a/b in patients with low plasma HDL-C levels.Methods. A total of 42 participating patients with low baseline levels of HDL cholesterol were assigned to receive an XZK capsule, 600 mg twice daily for 6 months. The expression of circulating miR-33a/b was detected at baseline and after XZK therapy measured with quantitative reverse-transcription (RT) polymerase chain reaction (PCR).Results. The mean (SD) HDL-C level after XZK treatment was 1.19 (0.13) mmol/L, representing an increase of 11.2% from baseline (P<0.001). Q-PCR analysis of plasma miRNAs revealed an increase in relative miR-33a/b expression with XZK treatment. The miR-33a expression was raised from 0.81 to 1.73 (P=0.012); miR-33b expression was increased from 1.2 to 2.75 (P<0.001). The changes of miR-33a and miR-33b were inversely related to the posttreatment LDL-C levels (r=-0.37,P=0.019;r=-0.33,P=0.035, resp.).Conclusion. In patients with low HDL-C levels, XZK therapy raised plasma levels of miR-33a and miR-33b, which may inhibit cellular cholesterol export and limit the HDL-raising effect of XZK.

scholarly journals Polymerase Chain Reaction and blood culture for diagnosis of canine sepsis

2018 ◽  
Vol 48 (6) ◽  
Author(s):  
Marcelo Marques da Silveira ◽  
Stéfhano Luis Cândido ◽  
Karin Rinaldi dos Santos ◽  
Maerle Oliveira Maia ◽  
Roberto Lopes de Souza ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Blood Culture ◽  
Diagnostic Value ◽  
Turnaround Time ◽  
Diagnostic Techniques ◽  
Pcr Analysis ◽  
Chain Reaction ◽  
Suspected Sepsis ◽  
Fungal Dna ◽  
Polymerase Chain

ABSTRACT: Sepsis is characterized by the presence of organ dysfunction secondary to the dysregulated systemic inflammatory response associated with an infection, and has high mortality rates. Traditional diagnostic techniques based on non-microbiological isolation are time-consuming and may delay treatment. Thus, this study aimed to compare bacterial and fungal broad-range polymerase chain reaction (PCR) and blood culture for diagnosis of sepsis in dogs. Blood samples from 88 dogs with suspected sepsis were analyzed by blood culture, and PCR to detect bacterial and fungal DNA. On blood culture, 20 (22.7%) samples tested positive for bacterial isolates; however, none tested positive for fungi. Through PCR analysis, bacterial DNA was detected in 46 (52.3%) animals, whereas fungal DNA was present in one (1.1%) sample. Our results showed that PCR-based testing has important diagnostic value for canine blood infections because it has a shorter turnaround time and higher sensitivity than traditional blood culture.

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