Search results for "QPCR"

Trichoderma asperellum, Bacillus subtilis un to konsorcija ietekme uz augsnes mikroorganismiem

2019

Pētījuma mērķis bija noskaidrot, vai Trichoderma asperellum, Bacillus subtilis un to konsorcijs ietekmē augsnes mikrobiotu. Pētījumā eksperimentālie augsnes paraugi tika apstrādāti ar B. subtilis, T.asperellum un B. cinerea mikroorganismiem un to konsorcijiem. Izmantojot mikrobioloģiskās metodes, tika noteikts baktēriju un sēņu daudzums dažādi apstrādātās augsnēs. Ar Biolog EcoPlates metodi, noteikta mikroorganismu metaboliskā aktivitāte augsnē. Trichoderma ģints, Firmicutes, Gammaproteobacteria un Acidobacteria DNS koncentrācija augsnes paraugos tika noteikta ar molekulārajām metodēm. Tika veikta korelācijas analīze starp mikrobioloģisko un molekulāro un tikai starp dažādiem molekulāro rez…

MARTX Toxin in the Zoonotic Serovar of Vibrio vulnificus Triggers an Early Cytokine Storm in Mice

2017

Vibrio vulnificus biotype 2-serovar E is a zoonotic clonal complex that can cause death by sepsis in humans and fish. Unlike other biotypes, Bt2 produces a unique type of MARTXVv (Multifunctional-Autoprocessive-Repeats-in-Toxin; RtxA13), which is encoded by a gene duplicated in the pVvBt2 plasmid and chromosome II. In this work, we analyzed the activity of this toxin and its role in human sepsis by performing in vitro, ex vivo, and in vivo assays. First, we demonstrated that the ACD domain, present exclusively in this toxin variant, effectively has an actin-cross-linking activity. Second, we determined that the whole toxin caused death of human endotheliocytes and monocytes by lysis and apo…

Differential responses of bacterial and archaeal groups at high taxonomical ranks to soil management

2010

Little is known about abundances of the major bacterial taxa in agricultural soils and how they are affected by fertilization or other agricultural practices. Our aim was to determine the abundance and relative distribution of several bacterial phyla and one class, as well as the archaeal and crenarchaeal communities, and how they were affected by different fertilization regimes to examine whether specific responses of microorganisms could be identified at these high taxonomic ranks. We used real-time PCR with taxa specific primers to quantify the abundance of the Actinobacteria, Acidobacteria, Bacteriodetes, Firmicutes, Gemmatimonadetes, Verrucomicrobia, Alphaproteobacteria and Crenarchaeo…

Detection of Parietaria Mottle Virus by RT-qPCR: An Emerging Virus Native of Mediterranean Area That Undermine Tomato and Pepper Production in Southe…

2021

Parietaria mottle virus (PMoV) is considered an emerging virus in many countries of the Mediterranean basin, especially on tomato and pepper crops. Symptoms on tomato leaves and fruits can be easily confused with those induced by cucumber mosaic virus (CMV) with necrogenic satellite RNA (CMV-satRNA), tomato spotted wilt virus (TSWV) or tomato mosaic virus (ToMV). Mixed infection of these viruses has been also reported in some tomato cultivars, with an increase in the complexity of the symptoms and severity of the disease. Although a specific serum and riboprobes have been produced, nowadays no sensitive diagnostic methods are available for the rapid PMoV detection. Here, we have developed a…

Potencial uso de la tecnología basada en microARNs para el diagnóstico y pronóstico de la escoliosis idiopática del adolescente

2021

Introducción: La escoliosis idiopática del adolescente (EIA) es una enfermedad que se caracteriza por una deformación tridimensional de la columna vertebral cuya etiología es aún desconocida. Actualmente no existe una explicación clara sobre la evolución de la enfermedad; El diagnóstico y seguimiento actual de la EIA se basa en el análisis de la curvatura de la columna mediante radiografía de rayos X, la cual está considerada como la técnica gold standard. Sin embargo, esta técnica no tiene valor pronóstico y las repetidas exposiciones han demostrado incrementar la incidencia de cáncer en la edad adulta. Es necesario, por tanto, el desarrollo de una nueva herramienta para el diagnóstico tem…

Changes in freshwater sediment microbial populations during fermentation of crude glycerol

2020

This work was supported by the Latvian Council of Science , project NN-CARMA, project No. lzp-2018/1-0194.

Hypoxia Positively Regulates the Expression of pH-Sensing G-Protein–Coupled Receptor OGR1 (GPR68)

2016

Background & Aims: A novel family of proton-sensing G-proteinâcoupled receptors, including ovarian cancer G-proteinâcoupled receptor 1 (OGR1) (GPR68) has been identified to play a role in pH homeostasis. Hypoxia is known to change tissue pH as a result of anaerobic glucose metabolism through the stabilization of hypoxia-inducible factor-1α. We investigated how hypoxia regulates the expression of OGR1 in the intestinal mucosa and associated cells. Methods: OGR1 expression in murine tumors, human colonic tissue, and myeloid cells was determined by quantitative reverse-transcription polymerase chain reaction. The influence of hypoxia on OGR1 expression was studied in monocytes/macrophages and…

A sensitive real-time RT-PCR reveals a high incidence of Southern tomato virus (STV) in Spanish tomato crops

2018

[EN] Southern tomato virus (STV) is a double-stranded RNA (dsRNA) virus belonging to genus Amalgavirus (family Amalgamaviridae). STV has been detected in tomato plants showing different symptoms although it has not been demonstrated that STV is the causal agent. To study the STV incidence and its pathogenic role, a sensitive and quantitative real-time reverse transcription-polymerase chain reaction assay (RT-qPCR) was developed. The standard curve perfonned with viral RNA transcripts allowed a wide dynamic range for STV quantitation from 10(4) to 10(11) copies/ng of total RNA. STV detection by RT-qPCR was 10(2)-fold more sensitive than conventional RT-PCR or RT-LAMP and 10(4)-fold more sens…

Bias of library preparation for virome characterization in untreated and treated wastewaters

2020

The use of metagenomics for virome characterization and its implementation for wastewater analyses, including wastewater-based epidemiology, has increased in the last years. However, the lack of standardized methods can led to highly different results. The aim of this work was to analyze virome profiles in upstream and downstream wastewater samples collected from four wastewater treatment plants (WWTPs) using two different library preparation kits. Viral particles were enriched from wastewater concentrates using a filtration and nuclease digestion procedure prior to total nucleic acid (NA) extraction. Sequencing was performed using the ScriptSeq v2 RNA-Seq (LS) and the NEBNext Ultra II RNA …

Molecular analyses on HPV infections in semen

Introduction: Human Papillomaviruses (HPVs) are non-enveloped double stranded DNA viruses classified in different genera and several different genotypes. Depending on the genotypes, HPV infections can be asymptomatic or can cause from warts to malignant tumours. Some genotypes, such as HPV18 and HPV16, are considered high risk (HR) HPVs and they are the major cause of cervical cancer. HPV is highly tissue-tropic and infects epithelial cells, but it can also binds other cell types. It is known that HPV can be found in semen but its effects on spermatozoa and male reproductive system are not completely clarified. Materials and Methods: In order to study the HPV infection in semen, we develope…