The current study evaluated the synergistic role of Plant growth promoting rhizobacteria (PGPR), Pseudomonas aeruginosa and Burkholderia gladioli on different physiological, biochemical and molecular ...activities of 10-days old Solanum lycopersicum seedlings under Cd stress. Cd toxicity altered the levels of phenolic compounds (total phenols (30.2%), flavonoids (92.7%), anthocyanin (59.5%), polyphenols (368.7%)), osmolytes (total osmolytes (10.3%), total carbohydrates (94%), reducing sugars (64.5%), trehalose (112.5%), glycine betaine (59%), proline (54.8%), and free amino acids (63%)), and organic acids in S. lycopersicum seedlings. Inoculation of P. aeruginosa and B. gladioli alleviated Cd-induced toxicity, which was manifested through enhanced phenolic compound levels and osmolytes. Additionally, the levels of low molecular weight organic acids (fumaric acid, malic acid, succinic acid, and citric acid) were also elevated. The expression of genes encoding enzymes for phenols and organic acid metabolism were also studied to be modulated that included CHS (chalcone synthase; 138.4%), PAL (phenylalanine ammonia lyase; 206.7%), CS (citrate synthase; 61.3%), SUCLG1 (succinyl Co-A ligase; 33.6%), SDH (succinate dehydrogenase; 23.2%), FH (fumarate hydratase; 12.4%), and MS (malate synthase; 41.2%) and found to be upregulated in seedlings inoculated independently with P. aeruginosa and B. gladioli. The results provide insights into the role of micro-organisms in alleviating Cd-induced physiological damage by altering levels of different metabolites.
•Cd induced toxicity in S. lycopersicum expressed in terms of phenolic compounds, osmoprotectants and organic acids.•Supplementation of Pseudomonas aeruginosa and Burkholderia gladioli alleviated Cd toxicity in S. lycopersicum seedlings.•Phenolic compounds, organic acids and osmoprotectatants were enhanced upon microbial inoculations in Cd stressed seedlings.•The genes encoding enzymes for phenol and organic acid metabolism were upregulated as observed by qRT-PCR.•The present study commends the use of PGPR against heavy metal tolerance in plants.
This study investigates GABAB protein expression and mRNA levels in three types of specimens. Two types of specimens from patients with temporal lobe epilepsy (TLE), secondary to hippocampal ...sclerosis, sclerotic hippocampal samples (TLE-HS), and tissue from the structurally preserved non-spiking ipsilateral superior temporal gyrus (TLE-STG) removed from the same patient during epilepsy surgery; and third specimen is hippocampal tissue from individuals with no history of epilepsy (post-mortem controls, PMC).
mRNA expression of GABAB subunits was quantified in TLE-HS, TLE-STG and PMC specimens by qRT-PCR. Qualitative and quantitative Western blot (WB) and immunohistochemistry techniques were employed to quantify and localize GABAB proteins subunits.
qRT-PCR data demonstrated an overall decrease of both GABAB1 isoforms in TLE-HS compared to TLE-STG. These results were mirrored by the WB findings. GABAB2 mRNA and protein were significantly reduced in TLE-HS samples compared to TLE-STG; however they appeared to be upregulated in TLE-HS compared to the PMC samples. Immunohistochemistry (IHC) showed that GABAB proteins were widely distributed in PMC and TLE-HS hippocampal sections with regional differences in the intensity of the signal. The higher expression of mature GABAB protein in TLE-HS than PMC is in agreement with previous studies. However, these findings could be due to post-mortem changes in PMC specimens. The TLE-STG samples examined here represent a better ‘control’ tissue compared to TLE-HS samples characterised by lower than expected GABAB expression. This interpretation provides a better explanation for previous functional studies suggesting reduced inhibition in TLE-HS tissue due to attenuated GABAB currents.
This article is part of the “Special Issue Dedicated to Norman G. Bowery”.
•This study investigates GABAB in three types of human specimens: two from patients with TLE-HS, TLE-STG and PMC.•This study investigates GABAB with three different quantitative techniques: RT-PCR, Western blot, and immunohistochemistry in human specimens.•The higher expression of mature GABAB protein in TLE-HS than PMC is in agreement with previous studies.•On the other hand, this study shows a statistically significant lower expression of GABAB2 in TLE-HS samples than in non-epileptogenic.•Downregulated GABAB2 in TLE-HS could represent one reason for impaired GABAergic inhibition in epileptogenic hippocampus.
Pilocytic astrocytoma (PAs) represents a significant portion of childhood primary brain tumors, with distinct histological and radiological features. The prevalence of KIAA1549::BRAF fusion in PAs ...has been well-established, this study aims to assess the prevalence of KIAA1549::BRAF fusions and explore their associations with tumor characteristics, radiological findings, and patient outcomes in PAs.
Histologically confirmed cases of PAs from a 5-year period were included in the study. Demographic, histopathological, and radiological data were collected, and immunohistochemistry was performed to characterize tumor markers. FISH and qRT-PCR assays were employed to detect KIAA1549::BRAF fusions. Statistical analyses were conducted to examine associations between fusion status and various other parameters.
Histological analysis revealed no significant differences in tumor features based on fusion status. However, younger age groups showed higher fusion prevalence. Radiologically, fusion-positive cases were distributed across different tumor subtypes SE, CWE and NCWE. Survival analysis did not demonstrate a significant impact of fusion status on overall survival, however most cases with recurrence and death harboured KIAA1549::BRAF fusion. Of 200 PAs, KIAA1549::BRAF fusions were detected in 64 % and 74 % of cases via qRT-PCR and FISH, respectively. Concordance between the two platforms was substantial (86 %).
KIAA1549::BRAF fusions are prevalent in PAs and can be reliably detected using both FISH and qRT-PCR assays. Cost considerations suggest qRT-PCR as a more economical option for fusion detection in routine clinical practice.
•The study examined KIAA1549-BRAF fusions in 200 cases and association with tumor features, radiology and patient outcomes•No significant tumor feature differences observed based on fusion status; higher prevalence in younger age groups.•Radiological features of NCWE and SE found dominantly associated with fusion-positive cases.•qRT-PCR and FISH detected fusions in 64% and 74% of cases,with 86% concordance and their pros and cons were discussed.
ZmNF-YA3 plays important roles in the regulation of photoperiod-dependent flowering time through interacting with ZmCO-like and ZMFPF1, and in drought-stress tolerance through JA and ABA signalling ...pathways in maize.
Abstract
Nuclear factor-Y (NF-Y) transcription factors are important regulators of several essential biological processes, including embryogenesis, drought resistance, meristem maintenance, and photoperiod-dependent flowering in Arabidopsis. However, the regulatory mechanisms of NF-Ys in maize (Zea mays) are not well understood yet. In this study, we identified an NF-Y transcription factor, ZmNF-YA3. Genome-wide analysis showed that ZmNF-YA3 bound to >6000 sites in the maize genome, 2259 of which are associated with genic sequences. ZmNF-YA3 was found to interact with CONSTANS-like (CO-like) and flowering promoting factor1 (FPF1) through yeast two-hybrid and bimolecular fluorescence complementation (BiFC) assays. Quantitative real-time reverse transcription-PCR (qRT-PCR) combined with yeast one-hybrid assay and EMSA suggested that NF-YA3 could promote early flowering by binding to the FLOWERING LOCUS T-like12 (FT-like12) promoter in maize. Morerover, we also showed that ZmNF-YA3 could improve drought and high-temperature tolerance through binding to the promoter regions of bHLH92, FAMA, and the jasmonic acid activator MYC4, respectively. These results contribute to a comprehensive understanding of the molecular mechanisms and regulatory networks of NF-Y transcription factors in regulating maize flowering time and stress response in maize.
Display omitted
•The expression of R2R3 MYB gene shed the light on its potential function.•The three approaches; qRT-PCR, semi-qRT-PCR and TEA analysis showed similar expression patterns.•The in ...vitro and in silico expression of R2R3 MYB provide molecular insights into fruit development.•Phylogeny analysis suggests a potential expression profiles in different important plants.
Assessment of gene expression is an essential and sensitive indicator of the biological activity and function of a certain gene. The expression profile of R2R3 MYB gene (Solyc02g067760) was investigated at ten development stages of tomato fruits (Lycopersicon esculentum) of Micro-Tom cultivar using three different approaches; quantitative RT-PCR, semi-quantitative RT-PCR and the promising Tomato Expression Atlas (TEA) bioinformatics tool. RNA was extracted from fruit tissues and used for synthesizing cDNA. Quantitative and semi-quantitative expression analyses of R2R3 MYB gene using specific primers consistently showed different expression patterns at the different development stages of tomato fruit. R2R3 MYB exhibited maximum expression at the anthesis stage, suggesting its potential involvement in the pollination and fruit set process. The gene expression was drastically reduced in all other studied stages after anthesis. In the same context, spatiotemporal expression of R2R3 MYB during the ten development stages of tomato fruit using TEA bioinformatics tool revealed a similar expression profile to that obtained by qRT-PCR and semi-qRT-PCR. Comparable results of the three approaches provide new insights into the assessment of tomato gene expression profiling in the presence of Actin gene (Solyc04g011500) as a normalizing expression reference. The obtained results confirm the potentially essential role of the R2R3 MYB in fruit set leading the future research into exploring techniques and stratify of overexpressing this gene in the relevant economic crops.
Objectives
This study was designed to investigate the role of circ_0078767/miR‐330‐3p/RASSF1A in non‐small‐cell lung cancer (NSCLC). Bioinformatic analysis was performed to screen for the ...differentially expressed genes in NSCLC tissues from adjacent lung tissues.
Materials and Methods
qRT‐PCR was used to detect the RNA expression of genes in cells and tissues, and Western blot was conducted to determine the protein levels of RASSF1A in tissues and cells. A miRanda algorithm was used to predict the targeted relationship among RNAs. A dual‐luciferase reporter gene assay was conducted to verify the targeted relationship. Flow cytometry was performed to investigate the effects of circ_0078767/miR‐330‐3p/RASSF1A on cell cycle progression and apoptosis. A CCK‐8 assay was conducted to explore the effects of circ_0078767/miR‐330‐3p/RASSF1A on cell proliferation. A transwell invasion assay was completed to study the effects of circ_0078767/miR‐330‐3p/RASSF1A on cell invasion. Lastly, an in vivo assay was conducted to investigate the effects of circ_0078767/miR‐330‐3p/RASSF1A on tumour development.
Results
Circ_0078767 and RASSF1A were downregulated, while miR‐330‐3p was upregulated in NSCLC tissues than that in adjacent tissues. miR‐330‐3p had a binding relationship with circ_0078767 and RASSF1A. The overexpression of circ_0078767 and RASSF1A or the underexpression of miR‐330‐3p significantly suppressed NSCLC cell viability, cell cycle progression and invasion while also significantly promoting cell apoptosis. Additionally, these modulations significantly suppressed in vivo tumour growth.
Conclusions
Circ_0078767 could suppress NSCLC progression by inhibiting miR‐330‐3p, which thereby increased RASSF1 levels.
The presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in wastewater samples has been documented in several countries. Wastewater-based epidemiology (WBE) is potentially ...effective for early warning of a COVID-19 outbreak. In this study, presence of SARS-CoV-2 RNA in wastewater samples was investigated and was compared with the number of the confirmed COVID-19 cases in the study area during COVID-19 outbreak in Japan. In total, 45 influent wastewater samples were collected from five wastewater treatment plants in Ishikawa and Toyama prefectures in Japan. During the study period, the numbers of confirmed COVID-19 cases in these prefectures increased from 0.3 and 0 to >20 per 100,000 people. SARS-CoV-2 ribonucleic acid (RNA) in the samples was detected using several PCR-based assays. Of the 45 samples, 21 were positive for SARS-CoV-2 according to at least one of the three quantitative RT-PCR assays. The detection frequency increased when the number of total confirmed SARS-CoV-2 cases in 100,000 people exceeded 10 in each prefecture; however, SARS-CoV-2 could also be detected at a low frequency even when the number was below 1.0. SARS-CoV-2 in wastewater could be detected in the early stage of the epidemic, even if the number of confirmed cases potentially underestimates the actual numbers of cases. This suggests that WBE approach can potentially act as an early warning of COVID-19 outbreaks in Japan.
Display omitted
•Presence of SARS-CoV-2 RNA in wastewater was studied in two prefectures in Japan.•At the start of the study, no cases of COVID-19 had been reported in the study area.•SARS-CoV-2 detection frequency increased along with the number of reported cases.•SARS-CoV-2 was detected even when the number of cases was <1.0 per 100,000 people.•The detection frequency remained high even after increase in the cases stopped.
PDF
