scholarly journals Sympatric Bat Species Prey Opportunistically on a Major Moth Pest of Pecans

2019 ◽  
Vol 11 (22) ◽  
pp. 6365
Author(s):  
Elizabeth C. Braun de Torrez ◽  
Veronica A. Brown ◽  
Gary F. McCracken ◽  
Thomas H. Kunz
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Single Species ◽  
Foraging Strategies ◽  
Lasiurus Borealis ◽  
Agricultural Matrix ◽  
Gene Copies ◽  
Polymerase Chain ◽  
Pest Suppression ◽  
Eastern Red Bat ◽  
Host Parasite

Native predators provide undervalued pest suppression services to agriculture. Studies of pest consumption by insectivorous bats tend to focus upon single species in large, centralized colonies, while bats dispersed in small groups within the agricultural matrix often go unnoticed. Pecan trees, Carya illinoinensis, and the destructive pecan nut casebearer (PNC) moth, Acrobasis nuxvorella, comprise a tightly linked host–parasite system in a widespread agroecosystem native to North America. Here we use a quantitative polymerase chain reaction (qPCR) assay of fecal DNA to document predation on PNC moths by an assemblage of sympatric bat species across episodic peaks in PNC abundance. Although five species of bats consume PNC moths, greater predation by a solitary tree-roosting bat (eastern red bat, Lasiurus borealis) than other species is suggested by a higher frequency of PNC occurrence and quantity of PNC gene copies in fecal samples. Consumption of PNC by bats during all documented peaks in moth activity suggests that predation pressure occurs throughout the PNC season. Our results highlight the need to consider multi-species assemblages and different foraging strategies when assessing pest suppression services, particularly in agroforestry or tree crops. Assessing the diet of only common or easily captured species limits our ability to accurately document pest consumption by bats.

scholarly journals Eliminating false positives in a qPCR assay for the detection of the uidA gene in Escherichia coli

2013 ◽  
Vol 11 (3) ◽  
pp. 382-386 ◽  
Author(s):  
Richard Kibbee ◽  
Natalie Linklater ◽  
Banu Örmeci
Keyword(s):  
Escherichia Coli ◽  
Quantitative Polymerase Chain Reaction ◽  
Qpcr Assay ◽  
Uida Gene ◽  
Chain Reaction ◽  
Taq Polymerase ◽  
E Coli ◽  
Gene Copies ◽  
Polymerase Chain ◽  
Low Levels

Due to contaminant Escherichia coli DNA present in recombinant Taq polymerase reagents, it is not possible to reliably detect low levels of E. coli in samples using the quantitative polymerase chain reaction (qPCR) assay. Native Taq polymerase was successfully used in this study to detect five uidA gene copies (5 fg of genomic DNA) of the uidA gene.

scholarly journals Effect of D-Mannose on Gene Expression of Neuraminidase Produced from Different Clinical Isolates of Pseudomonas aeruginosa

2019 ◽  
Vol 16 (2) ◽  
pp. 0291
Author(s):  
Shehab Et al.
Keyword(s):  
Gene Expression ◽  
Pseudomonas Aeruginosa ◽  
Antibacterial Agents ◽  
Quantitative Polymerase Chain Reaction ◽  
Clinical Isolates ◽  
Chain Reaction ◽  
Gene Copies ◽  
Different Types ◽  
Polymerase Chain

The present work aimed to investigate the neuraminidase (nan1) gene expression in 32 different clinical isolates of Pseudomonas aeruginosa to explore the role of the enzyme in different types of infection and might give a better understanding of host cell-pathogens interaction. In addition, the effect of monosaccharide D-mannose on neuraminidase gene expression in eight isolates was studied by utilizing a reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The results demonstrated that the highest expression of nan1 gene was in otitis samples (208,913.81) which were significantly higher than that from other infections (P < 0.01). While, the concentrations of gene copies obtained from urine, sputum and burns samples were 93,535.34, 92,254.64 and 74,029.63respectively. While the least expression in wound samples (32,017.06). This suggests that neuraminidase in ear samples might be more virulent and invasive followed by that from urine, sputum, burns and wounds samples. The considerable interest of addition D-mannose significantly reduced the rate of neuraminidase activity reached fivefold in some isolates. This indicates that D-mannose down regulates nan1 gene expression. Hence, this sugar could be used in the development of potential new antibacterial agents where it acts as a competitive neuraminidase inhibitors.

scholarly journals Effect of D-Mannose on Gene Expression of Neuraminidase Produced from Different Clinical Isolates of Pseudomonas aeruginosa

2019 ◽  
Vol 16 (2) ◽  
pp. 0291
Author(s):  
Shehab Et al.
Keyword(s):  
Gene Expression ◽  
Pseudomonas Aeruginosa ◽  
Antibacterial Agents ◽  
Quantitative Polymerase Chain Reaction ◽  
Clinical Isolates ◽  
Chain Reaction ◽  
Gene Copies ◽  
Different Types ◽  
Polymerase Chain

The present work aimed to investigate the neuraminidase (nan1) gene expression in 32 different clinical isolates of Pseudomonas aeruginosa to explore the role of the enzyme in different types of infection and might give a better understanding of host cell-pathogens interaction. In addition, the effect of monosaccharide D-mannose on neuraminidase gene expression in eight isolates was studied by utilizing a reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The results demonstrated that the highest expression of nan1 gene was in otitis samples (208,913.81) which were significantly higher than that from other infections (P < 0.01). While, the concentrations of gene copies obtained from urine, sputum and burns samples were 93,535.34, 92,254.64 and 74,029.63respectively. While the least expression in wound samples (32,017.06). This suggests that neuraminidase in ear samples might be more virulent and invasive followed by that from urine, sputum, burns and wounds samples. The considerable interest of addition D-mannose significantly reduced the rate of neuraminidase activity reached fivefold in some isolates. This indicates that D-mannose down regulates nan1 gene expression. Hence, this sugar could be used in the development of potential new antibacterial agents where it acts as a competitive neuraminidase inhibitors.

Expression profiling of circulating miRNAs in mouse serum in response to Echinococcus multilocularis infection

Parasitology ◽  
2017 ◽  
Vol 144 (8) ◽  
pp. 1079-1087 ◽  
Author(s):  
XIAOLA GUO ◽  
YADONG ZHENG
Keyword(s):  
Echinococcus Multilocularis ◽  
Quantitative Polymerase Chain Reaction ◽  
Mouse Serum ◽  
Sequencing Data ◽  
Circulating Micrornas ◽  
Serum Mirnas ◽  
Helminthic Infection ◽  
Host Origin ◽  
Polymerase Chain ◽  
Host Parasite

SUMMARYEchinococcus multilocularis is a most pathogenic zoonotic tapeworm that causes devastating echinococcosis in both humans and animals. Circulating microRNAs (miRNAs) are stably existed in the serum/plasma of mammalian hosts during helminthic infection. In this study, we compared the host-circulating miRNA expression in the sera from the E. multilocularis-infected and uninfected mice. A total of 58 host-origin serum miRNAs were differentially expressed (2 ⩾ fold change, P < 0·05), of which 21 were upregulated and 37 were significantly downregulated. Consistent with the sequencing data, quantitative polymerase chain reaction (PCR) results showed that the expression levels of four miRNAs were elevated gradually and one decreased gradually at the E. multilocularis infection time points. Moreover, seven of E. multilocularis specific miRNAs were identified in the sera. Real-time PCR analyses further demonstrated that only two parasite-derived miRNAs (emu-miR-10 and emu-miR-227) were specifically amplified in all the sera from mice infected with E. multilocularis. These findings will be helpful to understand the roles of miRNAs in host–parasite interaction and to potentiate serum miRNAs as diagnostic targets for echinococcosis.

Increased copper levels inhibit denitrification in urban soils

2018 ◽  
Vol 109 (3-4) ◽  
pp. 421-427
Author(s):  
Shun LI ◽  
Xiaoru YANG ◽  
Daniel BUCHNER ◽  
Haitao WANG ◽  
Huijuan XU ◽  
...  
Keyword(s):  
Urban Soils ◽  
Anthropogenic Activities ◽  
Quantitative Polymerase Chain Reaction ◽  
Urban Ecosystems ◽  
Urban Park ◽  
Marker Genes ◽  
Copy Numbers ◽  
Gene Copies ◽  
Denitrification Activity ◽  
Polymerase Chain

ABSTRACTThe consequences of urbanisation for Earth's biogeochemical cycles are largely unexplored. Copper (Cu) in urban soils is being accumulated mainly due to anthropogenic activities under rapid urbanisation. The increasing Cu concentrations may contribute to altering soil nitrogen (N) cycling in urban ecosystems through modulating denitrification processes. This research aims to identify how Cu impacts urban soil denitrification functions and denitrifier abundance. An urban park soil with a background total Cu concentration of 7.9μgg–1 was incubated anaerobically with different Cu amendments (10, 20, 40, 80 and 160μg Cu g–1 soil), similar to prevalent Cu contents in urban soils. We evaluated the soil denitrification functions using the acetylene (C2H2) inhibition method and assessed the denitrifier abundance by quantitative polymerase chain reaction (qPCR) analyses of denitrifying marker genes (nirK, nirS and nosZ). At the function level, we observed that both the potential soil denitrification activity and the N2O emission rate due to denitrification were significantly (P<0.05) inhibited by Cu; even the lowest Cu addition (10μg Cu g–1 soil) drastically affected the denitrification function. Moreover, Cu significantly (P<0.05) decreased the abundance of nirK and nirS genes at the additions of 160μg Cu g–1 soil and 40μg Cu g–1 soil, respectively, whereas it had no clear impact on nosZ gene copies. Further correlation analyses revealed that the potential denitrification activity was positively correlated to the copy numbers of nirK and nirS genes, but it was not correlated to nosZ gene abundance. These findings indicate that Cu additions inhibited soil denitrification function and decreased denitrifier abundance in the investigated urban park soil. Our results suggest that Cu accumulation in urban soils, resulting from urbanisation, may generally influence denitrification in urban ecosystems.

scholarly journals Dynamics of Mixed–Candida Species Biofilms in Response to Antifungals

2017 ◽  
Vol 97 (1) ◽  
pp. 91-98 ◽  
Author(s):  
G. Vipulanandan ◽  
M. Herrera ◽  
N.P. Wiederhold ◽  
X. Li ◽  
J. Mintz ◽  
...  
Keyword(s):  
Amphotericin B ◽  
Candida Species ◽  
Fungal Pathogen ◽  
Quantitative Polymerase Chain Reaction ◽  
Single Species ◽  
Antifungal Treatment ◽  
Oral Infections ◽  
Human Fungal Pathogen ◽  
Polymerase Chain ◽  
Oral Colonization

Oral infections caused by Candida species, the most commonly isolated human fungal pathogen, are frequently associated with biofilms. Although Candida albicans is the predominant organism found in patients with oral thrush, a biofilm infection, there is an increasing incidence of oral colonization and infections caused by non- albicans Candida species, including C. glabrata, C. dubliniensis, and C. tropicalis, which are frequently more resistant to antifungal treatment. While single-species Candida biofilms have been well studied, considerably less is known about the dynamics of mixed– Candida species biofilms and how these dynamics are altered by antifungal treatment. To address these questions, we developed a quantitative polymerase chain reaction–based approach to determine the precise species composition of mixed– Candida species biofilms formed by clinical isolates and laboratory strains in the presence and absence of clinically relevant concentrations of 3 commonly used antifungals: fluconazole, caspofungin, and amphotericin B. In monospecies biofilms, fluconazole exposure favored growth of C. glabrata and C. tropicalis, while caspofungin generally favored significant growth of all species to a varying degree. Fluconazole was not effective against preformed mixed– Candida species biofilms while amphotericin B was potent. As a general trend, in mixed– Candida species biofilms, C. albicans lost dominance in the presence of antifungals. Interestingly, presence in mixed versus monospecies biofilms reduced susceptibility to amphotericin B for C. tropicalis and C. glabrata. Overall, our data suggest that antifungal treatment favors the growth of specific non- albicans Candida species in mixed– Candida species biofilms.

AN IMPROVED METHOD FOR INHIBITOR FREE NUCLEIC ACIDS FROM POLYPHENOL RICH LEAVES OF MUSA SPP

2017 ◽  
Vol 23 (1) ◽  
Author(s):  
N.NANDHA KUMAR ◽  
K. SOURIANATHA SUNDARAM ◽  
D. SUDHAKAR ◽  
K.K. KUMAR
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Proteinase K ◽  
Chain Reaction ◽  
Banana Plant ◽  
High Molecular Weight Dna ◽  
Musa Spp ◽  
Leaf Tissues ◽  
Polymerase Chain

Excessive presence of polysaccharides, polyphenol and secondary metabolites in banana plant affects the quality of DNA and it leads to difficult in isolating good quality of DNA. An optimized modified CTAB protocol for the isolation of high quality and quantity of DNA obtained from banana leaf tissues has been developed. In this protocol a slight increased salt (NaCl) concentration (2.0M) was used in the extraction buffer. Polyvinylpyrrolidone (PVP) and Octanol were used for the removal of polyphenols and polymerase chain reaction (PCR) inhibitors. Proteins like various enzymes were degraded by Proteinase K and removed by centrifugation from plant extract during the isolation process resulting in pure genomic DNA, ready to use in downstream applications including PCR, quantitative polymerase chain reaction (qPCR), ligation, restriction and sequencing. This protocol yielded a high molecular weight DNA isolated from polyphenols rich leaves of Musa spp which was free from contamination and colour. The average yields of total DNA from leaf ranged from 917.4 to 1860.9 ng/ìL. This modified CTAB protocol reported here is less time consuming 4-5h, reproducible and can be used for a broad spectrum of plant species which have polyphenol and polysaccharide compounds.

Pim-1 is up-regulated by constitutively activated FLT3 and plays a role in FLT3-mediated cell survival

Blood ◽  
2005 ◽  
Vol 105 (4) ◽  
pp. 1759-1767 ◽  
Author(s):  
Kyu-Tae Kim ◽  
Kristin Baird ◽  
Joon-Young Ahn ◽  
Paul Meltzer ◽  
Michael Lilly ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Quantitative Polymerase Chain Reaction ◽  
Colony Growth ◽  
Dominant Negative ◽  
Internal Tandem Duplication ◽  
Downstream Target ◽  
Before And After ◽  
Flt3 Expression ◽  
Polymerase Chain ◽  
Expression Microarrays

AbstractConstitutively activating internal tandem duplication (ITD) mutations of the receptor tyrosine kinase FLT3 (Fms-like tyrosine kinase 3) play an important role in leukemogenesis, and their presence is associated with poor prognosis in acute myeloid leukemia (AML). To better understand FLT3 signaling in leukemogenesis, we have examined the changes in gene expression induced by FLT3/ITD or constitutively activated wild-type FLT3 expression. Microarrays were used with RNA harvested before and after inhibition of FLT3 signaling. Pim-1 was found to be one of the most significantly down-regulated genes upon FLT3 inhibition. Pim-1 is a proto-oncogene and is known to be up-regulated by signal transducer and activator of transcription 5 (STAT5), which itself is a downstream target of FLT3 signaling. Quantitative polymerase chain reaction (QPCR) confirmed the microarray results and demonstrated approximately 10-fold decreases in Pim-1 expression in response to FLT3 inhibition. Pim-1 protein also decreased rapidly in parallel with decreasing autophosphorylation activity of FLT3. Enforced expression of either the 44-kDa or 33-kDa Pim-1 isotypes resulted in increased resistance to FLT3 inhibition-mediated cytotoxicity and apoptosis. In contrast, expression of a dominant-negative Pim-1 construct accelerated cytotoxicity in response to FLT3 inhibition and inhibited colony growth of FLT3/ITD-transformed BaF3 cells. These findings demonstrate that constitutively activated FLT3 signaling up-regulates Pim-1 expression in leukemia cells. This up-regulation contributes to the proliferative and antiapoptotic pathways induced by FLT3 signaling. (Blood. 2005;105: 1759-1767)

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