US
array(50) {
  ["SERVER_SOFTWARE"]=>
  string(6) "Apache"
  ["REQUEST_URI"]=>
  string(35) "/product/qscript-xlt-cdna-supermix/"
  ["PATH"]=>
  string(49) "/usr/local/sbin:/usr/local/bin:/usr/sbin:/usr/bin"
  ["PP_CUSTOM_PHP_INI"]=>
  string(48) "/var/www/vhosts/system/quantabio.com/etc/php.ini"
  ["PP_CUSTOM_PHP_CGI_INDEX"]=>
  string(19) "plesk-php74-fastcgi"
  ["SCRIPT_NAME"]=>
  string(10) "/index.php"
  ["QUERY_STRING"]=>
  string(0) ""
  ["REQUEST_METHOD"]=>
  string(3) "GET"
  ["SERVER_PROTOCOL"]=>
  string(8) "HTTP/1.1"
  ["GATEWAY_INTERFACE"]=>
  string(7) "CGI/1.1"
  ["REDIRECT_URL"]=>
  string(35) "/product/qscript-xlt-cdna-supermix/"
  ["REMOTE_PORT"]=>
  string(5) "52000"
  ["SCRIPT_FILENAME"]=>
  string(48) "/var/www/vhosts/quantabio.com/httpdocs/index.php"
  ["SERVER_ADMIN"]=>
  string(14) "root@localhost"
  ["CONTEXT_DOCUMENT_ROOT"]=>
  string(38) "/var/www/vhosts/quantabio.com/httpdocs"
  ["CONTEXT_PREFIX"]=>
  string(0) ""
  ["REQUEST_SCHEME"]=>
  string(5) "https"
  ["DOCUMENT_ROOT"]=>
  string(38) "/var/www/vhosts/quantabio.com/httpdocs"
  ["REMOTE_ADDR"]=>
  string(11) "23.20.20.52"
  ["SERVER_PORT"]=>
  string(3) "443"
  ["SERVER_ADDR"]=>
  string(13) "172.31.63.191"
  ["SERVER_NAME"]=>
  string(17) "www.quantabio.com"
  ["SERVER_SIGNATURE"]=>
  string(0) ""
  ["HTTP_ACCEPT_ENCODING"]=>
  string(7) "br,gzip"
  ["HTTP_ACCEPT_LANGUAGE"]=>
  string(14) "en-US,en;q=0.5"
  ["HTTP_ACCEPT"]=>
  string(63) "text/html,application/xhtml+xml,application/xml;q=0.9,*/*;q=0.8"
  ["HTTP_USER_AGENT"]=>
  string(40) "CCBot/2.0 (https://commoncrawl.org/faq/)"
  ["HTTP_X_SUCURI_COUNTRY"]=>
  string(2) "US"
  ["HTTP_X_SUCURI_CLIENTIP"]=>
  string(11) "23.20.20.52"
  ["HTTP_X_FORWARDED_PROTO"]=>
  string(5) "https"
  ["HTTP_CONNECTION"]=>
  string(5) "close"
  ["HTTP_X_FORWARDED_FOR"]=>
  string(11) "23.20.20.52"
  ["HTTP_X_REAL_IP"]=>
  string(13) "185.93.229.32"
  ["HTTP_HOST"]=>
  string(17) "www.quantabio.com"
  ["HTTPS"]=>
  string(2) "on"
  ["HTTP_AUTHORIZATION"]=>
  string(0) ""
  ["SCRIPT_URI"]=>
  string(60) "https://www.quantabio.com/product/qscript-xlt-cdna-supermix/"
  ["SCRIPT_URL"]=>
  string(35) "/product/qscript-xlt-cdna-supermix/"
  ["UNIQUE_ID"]=>
  string(27) "YfG42sIl-42ozGROHy2qrgAAAAQ"
  ["REDIRECT_STATUS"]=>
  string(3) "200"
  ["REDIRECT_HTTPS"]=>
  string(2) "on"
  ["REDIRECT_HTTP_AUTHORIZATION"]=>
  string(0) ""
  ["REDIRECT_SCRIPT_URI"]=>
  string(60) "https://www.quantabio.com/product/qscript-xlt-cdna-supermix/"
  ["REDIRECT_SCRIPT_URL"]=>
  string(35) "/product/qscript-xlt-cdna-supermix/"
  ["REDIRECT_UNIQUE_ID"]=>
  string(27) "YfG42sIl-42ozGROHy2qrgAAAAQ"
  ["FCGI_ROLE"]=>
  string(9) "RESPONDER"
  ["PHP_SELF"]=>
  string(10) "/index.php"
  ["REQUEST_TIME_FLOAT"]=>
  float(1643231450.0399)
  ["REQUEST_TIME"]=>
  int(1643231450)
  ["SUCURIREAL_REMOTE_ADDR"]=>
  string(13) "185.93.229.32"
}

qScript XLT cDNA SuperMix

Premium Performance with SuperMix Benefits
Features & Benefits
  • Stabilized 5X concentrated master mix simplifies reaction assembly and improves accuracy by minimizing pipetting steps
  • Optimized, double pre-primed SuperMix provides equal representation of 5′ and 3′ RNA sequences ≤ 1kb for quantitative and conventional two-step RT-PCR
  • Improved RNA template binding affinity for increased yields with problematic sequences or complex secondary structure
  • Perform more qPCR assays from limiting amounts of RNA with 2-3 fold improved cDNA yields over older enzyme technologies
  • Broad linear dynamic detection range spanning up to 108 fold

 

qScript XLT cDNA SuperMix is intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.

Product
Kit Size
Order Info
Product
Kit Size
Order Info
qScript XLT cDNA SuperMix
Request Sample
Kit Size:
Order Info:
25 x 20 μL rxns (1 x 100 µL)
100 x 20 μL rxns (1 x 400 µL)
500 x 20 μL rxns (2 x 1000 µL)

Description

qScript XLT cDNA SuperMix is a next-generation tool for first-strand cDNA synthesis, providing a highly sensitive and easy-to-use solution for two-step RT-PCR and RT-qPCR. qScript XLT is an engineered M-MLV reverse transcriptase mutant with reduced RNase H activity and improved yield and stability at higher temperatures. Combined with a precise mixture of reaction components, this SuperMix enables superior results over a wide dynamic range of input RNA, with up to 8-fold higher sensitivity than our previous qScript cDNA SuperMix cDNA synthesis kits, which utilize an engineered RNase H(+) reverse transcriptase mutant.

Details

Details

Contents

Single-tube, 5X concentrated reagent containing:

  • Reaction buffer with optimized concentrations of molecular-grade MgCl2, dATP, dCTP, dGTP, dTTP
  • Recombinant ribonuclease inhibitor protein (RIP)
  • qScript XLT reverse transcriptase
  • Titrated concentrations of random hexamer and oligo(dT) primer
  • Proprietary enzyme stabilizers and performance-enhancing additives
Customer Testimonials

Customer Testimonials

qScript XLT cDNA SuperMix

"qScript XLT gave us a very clean cDNA that upon qPCR allowed a detection of the gene expression 3-8 cycles earlier than in our current RT."

Ernesto D. | UCSF
qScript XLT cDNA SuperMix

"The cDNA was easy to make and prep was of good quality. I have used 1 ug of total RNA as starting material."

Researcher | University of New Mexico

Details

Contents

Single-tube, 5X concentrated reagent containing:

  • Reaction buffer with optimized concentrations of molecular-grade MgCl2, dATP, dCTP, dGTP, dTTP
  • Recombinant ribonuclease inhibitor protein (RIP)
  • qScript XLT reverse transcriptase
  • Titrated concentrations of random hexamer and oligo(dT) primer
  • Proprietary enzyme stabilizers and performance-enhancing additives

Performance Data

Resources

Publications

The Betacoronavirus PHEV Replicates and Disrupts the Respiratory Epithelia and Upregulates Key Pattern Recognition Receptor Genes and Downstream Mediators, Including IL-8 and IFN-l
Rahul Nelli - 2021
Abstract
The upper respiratory tract is the primary site of infection by porcine hemagglutinating encephalomyelitis virus (PHEV). In this study, primary porcine respiratory epithelial cells (PRECs) were cultured in an air-liquid interface (ALI) to differentiate into a pseudostratified columnar epithelium, proliferative basal cells, M cells, ciliated cells, and mucus-secreting goblet cells. ALI-PRECs recreates a cell culture environment morphologically and functionally more representative of the epithelial lining of the swine trachea than traditional culture systems. PHEV replicated actively in this environment, inducing cytopathic changes and progressive disruption of the mucociliary apparatus. The innate immunity against PHEV was comparatively evaluated in ALI-PREC cultures and tracheal tissue sections derived from the same cesarean-derived, colostrum-deprived (CDCD) neonatal donor pigs. Increased expression levels of TLR3 and/or TLR7, RIG1, and MyD88 genes were detected in response to infection, resulting in the transcriptional upregulation of IFN-l1 in both ALI-PREC cultures and tracheal epithelia. IFN-l1 triggered the upregulation of the transcription factor STAT1, which in turn induced the expression of the antiviral IFN-stimulated genes OAS1 and Mx1. No significant modulation of the major proinflammatory cytokines interleukin-1b (IL-1b), IL-6, and tumor necrosis factor alpha (TNF-a) was detected in response to PHEV infection. However, a significant upregulation of different chemokines was observed in ALI-PREC cultures (CCL2, CCL5, CXCL8, and CXCL10) and tracheal epithelium (CXCL8 and CXCL10). This study shed light on the molecular mechanisms driving the innate immune response to PHEV at the airway epithelium, underscoring the important role of respiratory epithelial cells in the maintenance of respiratory homeostasis and on the initiation, resolution, and outcome of the infectious process. IMPORTANCE The neurotropic betacoronavirus porcine hemagglutinating encephalomyelitis virus (PHEV) primarily infects and replicates in the swine upper respiratory tract, causing vomiting and wasting disease and/or encephalomyelitis in suckling pigs. This study investigated the modulation of key early innate immune genes at the respiratory epithelia in vivo, on tracheal tissue sections from experimentally infected pigs, and in vitro, on air-liquid interface porcine respiratory cell cultures. The results from the study underscore the important role of respiratory epithelial cells in maintaining respiratory homeostasis and on the initiation, resolution, and outcome of the PHEV infectious process
The effects of palbociclib in combination with radiation in preclinical models of aggressive meningioma
Craig Horbinski - 2021
Abstract
Background. Meningiomas are the most common tumor arising within the cranium of adults. Despite surgical resection and radiotherapy, many meningiomas invade the brain, and many recur, often repeatedly. To date, no chemotherapy has proven effective against such tumors. Thus, there is an urgent need for chemotherapeutic options for treating meningiomas, especially those that enhance radiotherapy. Palbociclib is an inhibitor of cyclindependent kinases 4 and 6 that has been shown to enhance radiotherapy in preclinical models of other cancers, is well-tolerated in patients, and is used to treat malignancies elsewhere in the body. We, therefore, sought to determine its therapeutic potential in preclinical models of meningioma. Methods. Patient-derived meningioma cells were tested in vitro and in vivo with combinations of palbociclib and radiation. Outputs included cell viability, apoptosis, clonogenicity, engrafted mouse survival, and analysis of engrafted tumor tissues after therapy. Results. We found that palbociclib was highly potent against p16-deficient, Rb-intact CH157 and IOMM-Lee meningioma cells in vitro, but was ineffective against p16-intact, Rb-deficient SF8295 meningioma cells. Palbociclib also enhanced the in vitro efficacy of radiotherapy when used against p16-deficient meningioma, as indicated by cell viability and clonogenic assays. In vivo, the combination of palbociclib and radiation extended the survival of mice bearing orthotopic p16 deficient meningioma xenografts, relative to each as a monotherapy. Conclusions. These data suggest that palbociclib could be repurposed to treat patients with p16- deficient, Rb-intact meningiomas, and that a clinical trial in combination with radiation therapy merits consideration.
5-HT recruits distinct neurocircuits to inhibit hunger-driven and non-hunger-driven feeding
Yanlin He - 2021
Abstract
Obesity is primarily a consequence of consuming calories beyond energetic requirements, but underpinning drivers have not been fully defined. 5-Hydroxytryptamine (5-HT) neurons in the dorsal Raphe nucleus (5-HTDRN) regulate different types of feeding behavior, such as eating to cope with hunger or for pleasure. Here, we observed that activation of 5-HTDRN to hypothalamic arcuate nucleus (5-HTDRN → ARH) projections inhibits food intake driven by hunger via actions at ARH 5-HT2C and 5-HT1B receptors, whereas activation of 5-HTDRN to ventral tegmental area (5-HTDRN → VTA) projections inhibits non-hunger-driven feeding via actions at 5-HT2C receptors. Further, hunger-driven feeding gradually activates ARH-projecting 5-HTDRN neurons via inhibiting their responsiveness to inhibitory GABAergic inputs; non-hunger-driven feeding activates VTA-projecting 5-HTDRN neurons through reducing a potassium outward current. Thus, our results support a model whereby parallel circuits modulate feeding behavior either in response to hunger or to hunger-independent cues.
The Molecular Underpinning of Life-History Evolution: Roles of the IIS network and the Building of a Reptilian Model
Abby Elizabeth Beatty - 2021
Abstract
The Insulin and Insulin-like Signaling (IIS) network regulates cellular processes including pre- and post-natal growth, cellular development, wound healing, reproduction, and longevity. Despite their importance on the physiology of vertebrates, the study of the specific functions of the top regulators of the IIS network — insulin-like growth factors (IGFs) and IGF binding proteins (IGFBPs) has been mostly limited to a few model organisms, namely labrodents. My dissertation aims to build a foundation for the development of a reptilian model to study IIS in the context of early life growth and reproduction. Towards this aim, the chapters of my dissertation (1) demonstrate that the expression patterns of IGF1 and IGF2 seen in labrodents are atypical relative to the typical patterns across amniotic clades, including humans; (2) characterize the gene expression of IGFs and IGFBPs across tissues and developmental stages in a model reptile, the brown anole lizard (Anolis sagrei); (3) discover that forced investment in tail regeneration results in increased investment in reproduction in the brown anole; and (4) demonstrate how a CURE focused on novel IIS research can be an effective teaching tool in the undergraduate biology classroom.
TNF controls a speed-accuracy tradeoff in the apoptotic decision to restrict viral spread
Jennifer Oyler-Yaniv1 - 2020
Abstract
Early commitment to apoptosis is an important antiviral strategy. However, fast decisions that are based on limited evidence can be erroneous and cause unnecessary cell death and tissue damage. How cells optimize their decision making strategy to account for both speed and accuracy is unclear. Here we show that exposure to TNF, which is secreted by macrophages during viral infection, causes cells to change their decision strategy from “slow and accurate” to “fast and error-prone”. Mathematical modeling combined with experiments in cell culture and mouse corneas show that the regulation of the apoptotic decision strategy is critical to prevent HSV-1 spread. These findings demonstrate that immune regulation of cellular cognitive processes dynamically changes a tissues’ tolerance for self-damage, which is required to protect against viral spread.
Validation of suitable reference genes for normalization of quantitative reverse transcriptase- polymerase chain reaction in rice infected by Xanthomonas oryzae pv. oryzae
Carlo Magno Sagun - 2020
Abstract
Bacterial blight (BB) caused by Xanthomonas oryzae pv. oryzae (Xoo) is a costly disease in rice that threatens global rice production. Gene expression analysis by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) allows the study of the underlying mechanisms of both BB pathogenesis and resistance. In relative quantification, reference genes are often used to normalize the results to remove technical variations allowing the determination of true biological changes in a pilot experiment. However, variations in the expression of these reference genes can lead to erroneous and unreliable results. Thus, choosing the most stable reference genes for any specific experimental condition is of utmost importance in qRT-PCR experiments. Here, we used geNorm, NormFinder, Bestkeeper, Delta-Ct and RefFinder programs and/or methods to analyze the stability of the expression of eleven candidate reference genes namely: 18S ribosomal RNA (18S rRNA), Actin-1 (ACT1), ADP-Ribosylation Factor (ARF), Endothelial differentiation factor (Edf), eukaryotic Elongation Factor-1α (eEF-1α), eukaryotic Initiation Factor-4a (eIF-4a), Profilin 2 (Prof2), Nucleic Acid Binding Protein (NABP), Triosephosphate Isomerase (TI), Ubiquitin 5 (UBQ5) and Ubiquitin 10 (UBQ10) in cDNA samples from BB-susceptible and Xa21-mediated resistant rice cultivars collected at various times after Xoo inoculation. Under our experimental conditions, Edf and TI were the most stable reference genes while the common housekeeping genes 18S rRNA, and UBQ5 were among the least stable genes. Though using either Edf or TI as internal control is adequate for gene expression analysis, we suggest using both genes to normalize the data of qRT-PCR assays for rice subjected to Xoo inoculation.
HDAC7 regulates histone 3 lysine 27 acetylation and transcriptional activity at super-enhancer-associated genes in breast cancer stem cells
Corrado Caslini - 2019
Abstract
Chromatin regulation through histone modifications plays an essential role in coordinated expression of multiple genes. Alterations in chromatin induced by histone modifiers and readers regulate critical transcriptional programs involved in both normal development and tumor differentiation. Recently, we identified that histone deacetylases HDAC1 and HDAC7 are necessary to maintain cancer stem cells (CSCs) in both breast and ovarian tumors. Here, we sought to investigate the CSC-specific function of HDAC1 and HDAC7 mechanistically by using a stem-like breast cancer (BrCa) cell model BPLER and matched nonstem tumor cell (nsTC)-like HMLER, along with conventional BrCa cell lines with different CSC enrichment levels. We found that HDAC1 and HDAC3 inhibition or knockdown results in HDAC7 downregulation, which is associated with a decrease in histone 3 lysine 27 acetylation (H3K27ac) at transcription start sites (TSS) and super-enhancers (SEs) prominently in stem-like BrCa cells. Importantly, these changes in chromatin landscape also correlate with the repression of many SE-associated oncogenes, including c-MYC, CD44, CDKN1B, SLUG, VDR, SMAD3, VEGFA, and XBP1. In stem-like BrCa cells, HDAC7 binds near TSS and to SEs of these oncogenes where it appears to contribute to both H3K27ac and transcriptional regulation. These results suggest that HDAC7 inactivation, directly or through inhibition of HDAC1 and HDAC3, can result in the inhibition of the CSC phenotype by downregulating multiple SE-associated oncogenes. The CSC selective nature of this mechanism and the prospect of inhibiting multiple oncogenes simultaneously makes development of HDAC7 specific inhibitors a compelling objective.
Low‐Dose Pesticide Mixture Induces Accelerated Mesenchymal Stem Cell Aging In Vitro
Xavier Leveque - 2019
Abstract
The general population is chronically exposed to multiple environmental contaminants such as pesticides. We have previously demonstrated that human mesenchymal stem cells (MSCs) exposed in vitro to low doses of a mixture of seven common pesticides showed a permanent phenotype modification with a specific induction of an oxidative stress‐related senescence. Pesticide mixture also induced a shift in MSC differentiation toward adipogenesis. Thus, we hypothesized that common combination of pesticides may induce a premature cellular aging of adult MSCs. Our goal was to evaluate if the prolonged exposure to pesticide mixture could accelerate aging‐related markers and in particular deteriorate the immunosuppressive properties of MSCs. MSCs exposed to pesticide mixture, under long‐term culture and obtained from aging donor, were compared by bulk RNA sequencing analysis. Aging, senescence, and immunomodulatory markers were compared. The protein expression of cellular aging‐associated metabolic markers and immune function of MSCs were analyzed. Functional analysis of the secretome impacts on immunomodulatory properties of MSCs was realized after 21 days' exposure to pesticide mixture. The RNA sequencing analysis of MSCs exposed to pesticide showed some similarities with cells from prolonged culture, but also with the MSCs of an aged donor. Changes in the metabolic markers MDH1, GOT and SIRT3, as well as an alteration in the modulation of active T cells and modifications in cytokine production are all associated with cellular aging. A modified functional profile was found with similarities to aging process.
Serotonin2B receptors in the rat dorsal raphe nucleus exert a GABA-mediated tonic inhibitory control on serotonin neurons
Adeline Cathala - 2019
Abstract
The central serotonin2B receptor (5-HT2BR) is a well-established modulator of dopamine (DA) neuron activity in the rodent brain. Recent studies in rats have shown that the effect of 5-HT2BR antagonists on accumbal and medial prefrontal cortex (mPFC) DA outflow results from a primary action in the dorsal raphe nucleus (DRN), where they activate 5-HT neurons innervating the mPFC. Although the mechanisms underlying this interaction remain largely unknown, data in the literature suggest the involvement of DRN GABAergic interneurons in the control of 5-HT activity. The present study examined this hypothesis using in vivo (intracerebral microdialysis) and in vitro (immunohistochemistry coupled to reverse transcription-polymerase chain reaction) experimental approaches in rats. Intraperitoneal (0.16 mg/kg) or intra-DRN (1 μM) administration of the selective 5-HT2BR antagonist RS 127445 increased 5-HT outflow in both the DRN and the mPFC, these effects being prevented by the intra-DRN perfusion of the GABAA antagonist bicuculline (100 μM), as well as by the subcutaneous (0.16 mg/kg) or the intra-DRN (0.1 μM) administration of the selective 5-HT1AR antagonist WAY 100635. The increase in DRN 5-HT outflow induced by the intra-DRN administration of the selective 5-HT reuptake inhibitor citalopram (0.1 μM) was potentiated by the intra-DRN administration (0.5 μM) of RS 127445 only in the absence of bicuculline perfusion. Finally, in vitro experiments revealed the presence of the 5-HT2BR mRNA on DRN GABAergic interneurons. Altogether, these results show that, in the rat DRN, 5-HT2BRs are located on GABAergic interneurons, and exert a tonic inhibitory control on 5-HT neurons innervating the mPFC.
Stratifying nonfunctional pituitary adenomas into two groups distinguished by macrophage subtypes
Garima Yagnik - 2019
Abstract
Tumor-associated macrophages (TAMs) polarize to M1 and M2 subtypes exerting anti-tumoral and pro-tumoral effects, respectively. To date, little is known about TAMs, their subtypes, and their roles in non-functional pituitary adenomas (NFPAs). We performed flow cytometry on single cell suspensions from 16 NFPAs, revealing that CD11b+ myeloid cells comprise an average of 7.3% of cells in NFPAs (range = 0.5%–27.1%), with qPCR revealing most CD11b+ cells to be monocyte-derived TAMs rather than native microglia. The most CD11b-enriched NFPAs (10–27% CD11b+) were the most expansile (size>3.5 cm or MIB1>3%). Increasing CD11b+ fraction was associated with decreased M2 TAMs and increased M1 TAMs. All NFPAs with cavernous sinus invasion had M2/M1 gene expression ratios above one, while 80% of NFPAs without cavernous sinus invasion had M2/M1<1 (P = 0.02). Cultured M2 macrophages promoted greater invasion (P < 10-5) and proliferation (P = 0.03) of primary NFPA cultures than M1 macrophages in a manner inhibited by siRNA targeting S100A9 and EZH2, respectively. Primary NFPA cultures were of two types: some recruited more monocytes in an MCP-1-dependent manner and polarized these to M2 TAMs, while others recruited fewer monocytes and polarized them to M1 TAMS in a GM-CSF-dependent manner. These findings suggest that TAM recruitment and polarization into the pro-tumoral M2 subtype drives NFPA proliferation and invasion. Robust M2 TAM infiltrate may occur during an NFPA growth phase before self-regulating into a slower growth phase with fewer overall TAMs and M1 polarization. Analyses like these could generate immunomodulatory therapies for NFPAs.
Different bacterial and viral pathogens trigger distinct immune responses in a globally invasive ant
Philip J. Lester - 2019
Abstract
Invasive species populations periodically collapse from high to low abundance, sometimes even to extinction. Pathogens and the burden they place on invader immune systems have been hypothesised as a mechanism for these collapses. We examined the association of the bacterial pathogen (Pseudomonas spp.) and the viral community with immune gene expression in the globally invasive Argentine ant (Linepithema humile (Mayr)). RNA-seq analysis found evidence for 17 different viruses in Argentine ants from New Zealand, including three bacteriophages with one (Pseudomonas phage PS-1) likely to be attacking the bacterial host. Pathogen loads and prevalence varied immensely. Transcriptomic data showed that immune gene expression was consistent with respect to the viral classification of negative-sense, positive-sense and double-stranded RNA viruses. Genes that were the most strongly associated with the positive-sense RNA viruses such as the Linepithema humile virus 1 (LHUV-1) and the Deformed wing virus (DWV) were peptide recognition proteins assigned to the Toll and Imd pathways. We then used principal components analysis and regression modelling to determine how RT-qPCR derived immune gene expression levels were associated with viral and bacterial loads. Argentine ants mounted a substantial immune response to both Pseudomonas and LHUV-1 infections, involving almost all immune pathways. Other viruses including DWV and the Kashmir bee virus appeared to have much less immunological influence. Different pathogens were associated with varying immunological responses, which we hypothesize to interact with and influence the invasion dynamics of this species.
Single-cell RNA sequencing identifies TGF-β as a key regenerative cue following LPS-induced lung injury
Kent A. Riemondy - 2019
Abstract
Many lung diseases result from a failure of efficient regeneration of damaged alveolar epithelial cells (AECs) after lung injury. During regeneration, AEC2s proliferate to replace lost cells, after which proliferation halts and some AEC2s transdifferentiate into AEC1s to restore normal alveolar structure and function. Although the mechanisms underlying AEC2 proliferation have been studied, the mechanisms responsible for halting proliferation and inducing transdifferentiation are poorly understood. To identify candidate signaling pathways responsible for halting proliferation and inducing transdifferentiation, we performed single-cell RNA sequencing on AEC2s during regeneration in a murine model of lung injury induced by intratracheal LPS. Unsupervised clustering revealed distinct subpopulations of regenerating AEC2s: proliferating, cell cycle arrest, and transdifferentiating. Gene expression analysis of these transitional subpopulations revealed that TGF-β signaling was highly upregulated in the cell cycle arrest subpopulation and relatively downregulated in transdifferentiating cells. In cultured AEC2s, TGF-β was necessary for cell cycle arrest but impeded transdifferentiation. We conclude that during regeneration after LPS-induced lung injury, TGF-β is a critical signal halting AEC2 proliferation but must be inactivated to allow transdifferentiation. This study provides insight into the molecular mechanisms regulating alveolar regeneration and the pathogenesis of diseases resulting from a failure of regeneration.
Sesamin and sesamolin reduce amyloid-β toxicity in a transgenic Caenorhabditis elegans
Roongpetch Keowkase - 2018
Abstract
Alzheimer’s disease (AD) is a devastating neurodegenerative disease characterized by β-amyloid (Aβ) plaques in the brain. At the present, there is no approved drug with a proven disease-modifying effect. Sesame seed (Sesame indicum) has long been known as a healthy food in Southeast Asian countries. Sesame lignans obtained from sesame seed possess antioxidant property that exhibit a variety of beneficial effects in various models. The objective of this study was to investigate the protective effects of sesame lignans including sesamin, sesamolin, and sesamol against Aβ toxicity in Caenorhabditis elegans (C. elegans) model of Aβ toxicity and to address whether these sesame lignans have a positive effect on lifespan extension. A transgenic C. elegans expressing human Aβ was used to investigate protective effects of sesame lignans against Aβ toxicity. Sesamin and sesamolin significantly alleviated Aβ-induced paralysis. The real-time PCR revealed that both sesamin and sesamolin did not affect the expression of Aβ transgene. However, we found that only sesamin inhibited Aβ oligomerization. These findings demonstrated that, among three sesame lignans tested, sesamin protected against Aβ toxicity by reducing toxic Aβ oligomers. Sesamin and sesamolin also significantly improved Aβ-induced defect in chemotaxis behavior and reversed the defect to normal. Moreover, sesamin prolonged median and mean lifespan of the wild type worm. On the other hand, sesamolin and sesamol failed to extend lifespan. These results offer valuable evidence for the future use of sesamin in the development of agents for the treatment of AD. It is also worth investigating the structure-activity relationship of lignan-related structures and their anti-Aβ toxicity activities in the future.
Differential gene expression to an LPS challenge in relation to exogenous corticosterone in the invasive cane toad (Rhinella marina)
Steven Gardner - 2018
Abstract
The cane toad (Rhinella marina) is an invasive amphibian in several parts of the world. Much of the research performed on assessing the dispersal potential of invasive species has focused immunity. Invaders are predicted to rely less on pro-inflammatory immunity, allowing them to allocate energy to dispersal. Elevated stress may play a role in regulation of immune responses used by invasive species. RNA sequencing of spleen tissue from cane toads subjected to an acute LPS challenge revealed genes coding for cytokines involved in typical innate responses such as phagocytic cell recruitment, extravasation, inflammation, and lymphocyte differentiation were significantly upregulated, while toads receiving transdermal application of corticosterone in addition to an LPS injection showed downregulation of genes involved with cell mediated immunity. These results indicate hormonal changes associated with acute stress may alter investment into mounting cell-mediated or humoral responses while allowing for prolonged phagocytic innate responses in this invasive species.
Viral RNA load and histological changes in tissues following experimental infection with an arterivirus of possums (wobbly possum disease virus)
Julia Giles - 2018
Abstract
Tissues from Australian brushtail possums (Trichosurus vulpecula) that had been experimentally infected with wobbly possum disease (WPD) virus (WPDV) were examined to elucidate pathogenesis of WPDV infection. Mononuclear inflammatory cell infiltrates were present in livers, kidneys, salivary glands and brains of WPD-affected possums. Specific staining was detected by immunohistochemistry within macrophages in the livers and kidneys, and undefined cell types in the brains. The highest viral RNA load was found in macrophage-rich tissues. The detection of viral RNA in the salivary gland, serum, kidney, bladder and urine is compatible with transmission via close physical contact during encounters such as fighting or grooming, or by contact with an environment that has been contaminated with saliva or urine. Levels of viral RNA remained high in all tissues tested throughout the study, suggesting that on-going virus replication and evasion of the immune responses may be important in the pathogenesis of disease.
MPZL2 is a novel gene associated with autosomal recessive nonsyndromic moderate hearing loss
Guney Bademci - 2018
Abstract
While recent studies have revealed a substantial portion of the genes underlying human hearing loss, the extensive genetic landscape has not been completely explored. Here, we report a loss-of-function variant (c.72delA) in MPZL2 in three unrelated multiplex families from Turkey and Iran with autosomal recessive nonsyndromic hearing loss. The variant co-segregates with moderate sensorineural hearing loss in all three families. We show a shared haplotype flanking the variant in our families implicating a single founder. While rare in other populations, the allele frequency of the variant is ~ 0.004 in Ashkenazi Jews, suggesting that it may be an important cause of moderate hearing loss in that population. We show that Mpzl2 is expressed in mouse inner ear, and the protein localizes in the auditory inner and outer hair cells, with an asymmetric subcellular localization. We thus present MPZL2 as a novel gene associated with sensorineural hearing loss.
The origins of global invasions of the German wasp (Vespula germanica) and its infection with four honey bee viruses
Evan C. Brenton-Rule - 2018
Abstract
A successful control or eradication programme using biological control or genetically-mediated methods requires knowledge of the origin and the extent of wasp genetic diversity. Mitochondrial DNA variation in the native and invaded range of the social wasp Vespula germanica was used to examine intra-specific genetic variation and invasive source populations. We also examined wasps for the presence of four viruses found in honey bees: Acute bee paralysis virus, Deformed wing virus, Israeli acute paralysis virus and Kashmir bee virus. German wasps showed reduced genetic diversity in the invaded range compared to that of their native range. Populations in the introduced range are likely to have arrived from different source populations. All four viral honey bee pathogens were found in V. germanica, although they varied in their distribution and strain. Multiple introductions of German wasps have occurred for most invaded regions, though some populations are genetically homogenous. The differing locations of origin will guide researchers searching for biocontrol agents and the reduced genetic diversity may make these wasps a potentially viable target for control via gene drives.
The splicing factor transformer2 (tra2) functions in the Drosophila fat body to regulate lipid storage
Cezary Mikoluk - 2018
Abstract
Excess nutrients are stored as triglycerides mainly in the adipose tissue of an animal and these triglycerides are located in structures called lipid droplets. Previous genome-wide RNAi screens in Drosophila cells identified splicing factors as playing a role in lipid droplet formation. Our lab has recently identified the SR protein, 9G8, as an important factor in fat storage as decreasing its levels results in augmented triglyceride storage in the fat body. Previous in vitro studies have implicated 9G8 in the regulation of splicing of the sex determination gene doublesex (dsx) by binding to transformer (tra) and transformer2 (tra2); however, any function of these sex determination proteins in regulating metabolism is unknown. In this study, we have uncovered a role of tra2 to regulate fat storage in vivo. Inducing tra2dsRNA in the adult fat body resulted in an increase in triglyceride levels but had no effect on glycogen storage. Consistent with the triglyceride phenotype, tra2 knockdown flies lived longer under starvation conditions. In addition, this increase in triglycerides is due to more fat storage per cell and not an increase in the number of fat cells. Interestingly, the splicing of CPT1, an enzyme involved in the breakdown of lipids, was altered in flies with decreased tra2. The less-catalytically active isoform of CPT1 accumulated in tra2dsRNA flies suggesting a decrease in lipid breakdown, which is consistent with the increased triglyceride levels observed in these flies. Together, these results suggest a link between mRNA splicing, sex determination and lipid metabolism and may provide insight into the mechanisms underlying tissue-specific splicing and nutrient storage.
Chronic BMY7378 treatment alters behavioral circadian rhythms
Jhenkruthi Vijaya Shankara - 2017
Abstract
The mammalian circadian clock is synchronized to the day:night cycle by light. Serotonin modulates the circadian effects of light, with agonists inhibiting response to light and antagonists enhancing responses to light. A special class of serotonergic compounds, the mixed 5-HT1A agonist/antagonists, potentiate light-induced phase advances by up to 400% when administered acutely. In this study, we examine the effects of one of these mixed 5-HT1A agonist/antagonists, BMY7378, when administered chronically. Thirty adult male hamsters were administered either vehicle or BMY7378 via surgically implanted osmotic minipumps over a period of 28 days. In a light:dark cycle, chronic BMY7378 advanced the phase angle of entrainment, prolonged the duration of the active phase, and attenuated the amplitude of the wheel running rhythm during the early night. In constant darkness, chronic treatment with BMY7378 significantly attenuated light-induced phase advances, but had no significant effect on light-induced phase delays. Non-photic phase shifts to daytime administration of a 5-HT1A/7 agonist were also attenuated by chronic BMY7378 treatment. qRT-PCR analysis revealed that chronic BMY7378 treatment upregulated mRNA for 5-HT1A and 5-HT1B receptors in the hypothalamus, and downregulated mRNA for 5-HT1A and monoamine oxidase-A in the brainstem. These results highlight adaptive changes of serotonin receptors in the brain to chronic treatment with BMY7378, and link such up- and down-regulation to changes in important circadian parameters. Such long-term changes to the circadian system should be considered when patients are treated chronically with drugs that alter serotonergic function. This article is protected by copyright. All rights reserved.
Treatment of Theiler’s virus-induced demyelinating disease with teriflunomide
Francesca Gilli - 2017
Abstract
Teriflunomide is an oral therapy approved for the treatment of relapsing remitting multiple sclerosis (MS), showing both anti-inflammatory and antiviral properties. Currently, it is uncertain whether one or both of these properties may explain teriflunomide’s beneficial effect in MS. Thus, to learn more about its mechanisms of action, we evaluated the effect of teriflunomide in the Theiler’s encephalomyelitis virus-induced demyelinating disease (TMEV-IDD) model, which is both a viral infection and an excellent model of the progressive disability of MS. We assessed the effects of the treatment on central nervous system (CNS) viral load, intrathecal immune response, and progressive neurological disability in mice intracranially infected with TMEV. In the TMEV-IDD model, we showed that teriflunomide has both anti-inflammatory and antiviral properties, but there seemed to be no impact on disability progression and intrathecal antibody production. Notably, benefits in TMEV-IDD were mostly mediated by effects on various cytokines produced in the CNS. Perhaps the most interesting result of the study has been teriflunomide’s antiviral activity in the CNS, indicating it may have a role as an antiviral prophylactic and therapeutic compound for CNS viral infections.
Biopsy-derived intestinal epithelial cell cultures for pathway based stratification of patients with inflammatory bowel disease
Wiebe Vanhove - 2017
Abstract
BackgroundEndoplasmic reticulum stress was shown to be pivotal in the pathogenesis of inflammatory bowel disease. Despite progress in IBD drug development, not more than one third of patients achieve steroid-free remission and mucosal healing with current therapies. Furthermore, patient stratification tools for therapy selection are lacking. We aimed to identify and quantify epithelial ER stress in a patient-specific manner in an attempt towards personalized therapy.MethodsA biopsy-derived intestinal epithelial cell culture system was developed and characterized. ER stress was induced by thapsigargin and quantified with a BiP ELISA on cell lysates from 35 patients with known genotypes who were grouped based on the number of IBD-associated ER stress and autophagy risk alleles.ResultsThe epithelial character of the cells was confirmed by E-cadherin, ZO-1 and MUC2 staining and CK-18,CK-20 and LGR5 gene expression. Patients with 3 risk alleles had higher median epithelial BiP-induction (vs. untreated) levels compared to patients with 1 or 2 risk alleles (p=0.026 and 0.043, respectively). When autophagy risk alleles were included and patients were stratified in genetic risk quartiles, patients in Q2, Q3 and Q4 had significantly higher ER stress (BiP) when compared to Q1 (p=0.034, 0.040 and 0.034, respectively).ConclusionWe developed and validated an ex vivo intestinal epithelial cell culture system and showed that patients with more ER stress and autophagy risk alleles have augmented epithelial ER stress responses. We thus presented a personalized approach whereby patient-specific defects can be identified which in turn could help in selecting tailored therapies.
Fetal sex alters maternal anti-Mullerian hormone during pregnancy in cattle
Anja Stojsin-Carter - 2017
Abstract
Anti-Mullerian hormone (AMH) is expressed by both male and female fetuses during mammalian development, with males expressing AMH earlier and at significantly higher concentration. The aim of the current study was to explore the potential impact of pregnancy and fetal sex on maternal AMH and to determine if plasma (Pl) AMH or placenta intercotyledonary membrane and cotyledonary AMH receptor 2 (AMHR2) mRNA expression differ in pregnant cows carrying male vs. female fetuses. AMH levels in blood were measured using a bovine optimized ELISA kit. Cows pregnant with a male fetus were observed to have a significantly greater difference in Pl AMH between day 35 and 135 of gestation. Average fetal AMH level between 54 and 220 days of gestation was also observed to be significantly higher in male vs. female fetuses. Intercotyledonary membranes and cotyledons were found to express AMHR2 between days 38 and 80 of gestation at similar levels in both fetal sexes. These findings support the hypothesis that fetal sex alters maternal Pl AMH during pregnancy in cattle.
Helicobacter pylori infection perturbs iron homeostasis in gastric epithelial cells
Sebastian E. Flores - 2017
Abstract
The iron deficiency anaemia that often accompanies infection with Helicobacter pylori may reflect increased uptake of iron into gastric epithelial cells. Here we show an infection-associated increase in total intracellular iron levels was associated with the redistribution of the transferrin receptor from the cell cytosol to the cell surface, and with increased levels of ferritin, an intracellular iron storage protein that corresponded with a significant increase in lysosomal stores of labile iron. In contrast, the pool of cytosolic labile iron was significantly decreased in infected cells. These changes in intracellular iron distribution were associated with the uptake and trafficking of H. pylori through the cells, and enhanced in strains capable of expressing the cagA virulence gene. We speculate that degradation of lysosomal ferritin may facilitate H. pylori pathogenesis, in addition to contributing to bacterial persistence in the human stomach.
AR-V7 in Peripheral Whole Blood of Patients with Castration-resistant Prostate Cancer: Association with Treatment-specific Outcome Under Abiraterone and Enzalutamide
Anna Katharina Seitz - 2017
Abstract
Background It has been demonstrated that androgen receptor splice variant 7 (AR-V7) expression in circulating tumor cells (CTCs) predicts poor treatment response in metastatic castration-resistant prostate cancer (mCRPC) patients treated with abiraterone or enzalutamide. Objective To develop a practical and robust liquid profiling approach for direct quantification of AR-V7 in peripheral whole blood without the need for CTC capture and to determine its potential for predicting treatment response in mCRPC patients. Design, setting, and participants Whole blood samples from a prospective biorepository of 85 mCRPC patients before treatment initiation with abiraterone (n = 56) or enzalutamide (n = 29) were analyzed via droplet digital polymerase chain reaction. Outcome measurements and statistical analysis The association of AR-V7 status with prostate-specific antigen (PSA) response defined by PSA decline ≥50% and with PSA–progression-free survival (PSA-PFS), clinical PFS, and overall survival (OS) was assessed. Results and limitations High AR-V7 expression levels in whole blood were detectable in 18% (15/85) of patients. No patient with high AR-V7 expression achieved a PSA response, and AR-V7 status was an independent predictor of PSA response in multivariable logistic regression analysis (p = 0.03). High AR-V7 expression was associated with shorter PSA-PFS (median 2.4 vs 3.7 mo; p < 0.001), shorter clinical PFS (median 2.7 vs 5.5 mo; p < 0.001), and shorter OS (median 4.0 vs. 13.9 mo; p < 0.001). On multivariable Cox regression analysis, high AR-V7 expression remained an independent predictor of shorter PSA-PFS (hazard ratio [HR] 7.0, 95% confidence interval [CI] 2.3–20.7; p < 0.001), shorter clinical PFS (HR 2.3, 95% CI 1.1–4.9; p = 0.02), and shorter OS (HR 3.0, 95% CI 1.4–6.3; p = 0.005). Conclusions Testing of AR-V7 mRNA levels in whole blood is a simple and promising approach to predict poor treatment outcome in mCRPC patients receiving abiraterone or enzalutamide. Patient summary We established a method for determining AR-V7 status in whole blood. This test predicted treatment resistance in patients with metastatic castration-resistant prostate cancer undergoing treatment with abiraterone or enzalutamide. Prospective validation is needed before application to clinical practice.
Bone marrow mesenchymal stem cell-derived CD63+ exosomes transport Wnt3a exteriorly and enhance dermal fibroblast proliferation, migration and angiogenesis in vitro
Jeffrey D. McBride - 2017
Abstract
Wnts are secreted glycoproteins that regulate stem cell self-renewal, differentiation, and cell-to-cell communication during embryonic development and in adult tissues. Bone marrow mesenchymal stem cells (BM-MSCs) have been shown to stimulate dermis repair and regeneration; however, it is unclear how BM-MSCs may modulate downstream Wnt signaling. While recent reports implicate that Wnt ligands and Wnt messenger RNAs (such as Wnt4) exist within the interior compartment of exosomes, it has been debated whether or not Wnts exist on the exterior surface of exosomes to travel in the extracellular space. To help answer this question, we utilized flow cytometry of magnetic beads coated with anti-CD63 antibodies, and found for the first time, that Wnt3a protein is detectable exteriorly on CD63+ exosomes derived from BM-MSCs over-secreting Wnt3a into serum-free conditioned media (Wnt3a CM). Our data suggests that CD63+ exosomes significantly help transport exterior Wnt3a signal to recipient cells to promote fibroblast and endothelial functions. During purification of exosomes, we unexpectedly found that use of ultracentrifugation alone significantly decreased the ability to detect exteriorly bound Wnt3a on CD63+ exosomes, however, polyethylene glycol-mediated exosome-enrichment prior to exosome-purification (with ultracentrifugation into a sucrose cushion) resulted in exosomes more likely to retain exterior Wnt3a detectability and downstream Wnt/beta-catenin activity. Our findings indicate the important role that purification methods may have on stem cell-derived Wnt-exosome activity in downstream assays. The ability for BM-MSC Wnt3a CM and exosomes to stimulate dermal fibroblast proliferation and migration, as well as endothelial angiogenesis in vitro, was significantly decreased after CD63+-exosome depletion or knockdown of Wnt coreceptor LRP6 in recipient cells, suggesting both are required for optimal Wnt-exosome activity in our system. Thus, BM-MSC-derived CD63+ exosomes are a significant carrier of exterior Wnt3a within high Wnt environments, resulting in downstream fibroblast proliferation, migration and angiogenesis in vitro.
Click here to see all Publications

Customer Testimonials

qScript XLT cDNA SuperMix

"qScript XLT gave us a very clean cDNA that upon qPCR allowed a detection of the gene expression 3-8 cycles earlier than in our current RT."

Ernesto D. | UCSF
qScript XLT cDNA SuperMix

"The cDNA was easy to make and prep was of good quality. I have used 1 ug of total RNA as starting material."

Researcher | University of New Mexico

Product Finder

Select Your Assay

Starting Template

Assay Format

Detection Chemistry

Multiplexing (more than 3 targets)

Is gene-specific priming (GSP) required?

What current Reverse Transcriptase or cDNA kit are you using?

Select the group which contains your real-time PCR cycler

  • Applied Biosystems 7500
  • Applied Biosystems 7500 Fast
  • Stratagene Mx3000P®
  • Stratagene Mx3005P™
  • Stratagene Mx4000™
  • Applied Biosystems ViiA 7
  • Applied Biosystems QuantStudio™
  • Agilent AriaMx
  • Douglas Scientific IntelliQube®
  • Applied Biosystems 5700
  • Applied Biosystems 7000
  • Applied Biosystems 7300
  • Applied Biosystems 7700
  • Applied Biosystems 7900
  • Applied Biosystems 7900HT
  • Applied Biosystems 7900 HT Fast
  • Applied Biosystems StepOne™
  • Applied Biosystems StepOnePlus™
  • Quantabio Q
  • BioRad CFX
  • Roche LightCycler 480
  • QIAGEN Rotor-Gene Q
  • Other
  • BioRad iCycler iQ™
  • BioRad MyiQ™
  • BioRad iQ™5

Choose your application from the categories below

Products