•RT-PCR demonstrated that egr-miR-71 and egr-let-7 were specifically amplified in all the plasma samples from the infected individuals with hydatid cyst.•The results of this study showed that two ...hydatid cyst derived miRNAs, egr-miR-71 and egr-miR-let-7, were significantly down-regulated after three and six months’ post-surgery and remove the cyst.
Currently, cystic echinococcosis (CE) follow-up is a serious concern among surgeons. MicroRNAs (miRNAs) are small, endogenous, non-coding RNAs which are present in human body fluids in a highly stable form. Recently, it is observed that Echinococcus granulosus expresses a large number of miRNAs in its developmental stages. The current study aimed at evaluating the capacity of parasitic miRNAs to serve as plasma biomarkers for hydatid cysts before and after CE surgery. Hydatidosis patients were identified using radiological and histopathological examinations. Following RNA extraction and cDNA synthesis, the expression levels of parasite-derived miRNAs including egr-miR-71 and egr-let-7 were quantitatively evaluated using real-time polymerase chain reaction (RT-PCR) in 30 hydatid cyst-infected individuals before surgery and an equal number of healthy controls. Then, three- and six-month follow-ups were performed after cystectomy. To analyze parasite-derived miRNAs, the relative fold change between uninfected and infected samples was determined and normalized to hsa-miR-16–5p as the housekeeping internal control. RT-PCR demonstrated that egr-miR-71 and egr-let-7 were specifically amplified in all the plasma samples from the infected individuals with hydatid cyst; yet they were significantly down-regulated at three and six months' post-surgery (P < 0.05). The egr-miR-71 had a higher level of expression in larval stage compared with egr-let-7. The results of the current study indicated that hydatid cyst-derived miRNAs including egr-miR-71 and egr-let-7 can be detected in human plasma. Considering the changes in the expression levels of these miRNAs after three and six months, it seems that these miRNAs, especially egr-miR-71, could serve as novel promising biomarkers for the early diagnosis and monitoring of hydatidosis.
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•Long-term ENF decreased the abundance of soil diazotrophs.•Long-term ENF decreased species richness and diversity of soil diazotrophs.•Soil pH and EC values were the main factors affecting soil ...diazotrophs.•The maize rhizosphere could enrich soil diazotrophs.•The diazotrophic bacterial variations under ENF might inhibit early maize growth.
Excessive nitrogen (N) fertilization (ENF) and low utilization efficiency of fertilizer-derived N in high-input, high-yield cropping systems are serious ecological and economic problems in China. To examine the effects of long-term ENF on soil diazotrophs in relation to maize growth, we performed a 5-year field trial whereby 0–300 kg N ha−1 were added to black soil under a maize monocropping system in Northeast China. At the end of the 5-year field trial, the abundance and community structure of diazotrophs in the bulk soil and rhizosphere were investigated at the maize jointing stage by real-time quantitative polymerase chain reaction (qRT-PCR) and high-throughput sequencing. The results showed that: 1) ENF (N300) limited maize growth in the early stages and did not increase grain yield compared with moderate N supplementation (N180); 2) ENF significantly reduced nifH copy number, species richness, and Shannon index values of soil diazotrophs by 53.7%–79.7%, 37.2%–47.6%, and 20.0%–31.6%, respectively, reduced the relative abundance of Burkholderia, and increased the relative abundance of Sphingobium; 3) ENF decreased soil pH and increased electrical conductivity, which were the main factors affecting soil diazotrophic community structure as determined by Mantel test; and 4) diazotrophs were 1.3–3.0 times more abundant in the rhizosphere than in bulk soil, but no differences in α-diversity and community composition of diazotrophs were observed between rhizosphere and bulk soil. Thus, long-term ENF has negative impacts on the soil diazotrophic community, inhibiting maize growth in the early stage. Split N fertilization or slow/controlled-released N fertilizer should be used to avoid the negative effects of ENF on the soil diazotrophic community structure and early maize growth.
Venous ulcers are the most common type of human chronic nonhealing wounds and are stalled in a constant and excessive inflammatory state. The molecular mechanisms underlying the chronic wound ...inflammation remain elusive. Moreover, little is known about the role of regulatory RNAs, such as microRNAs, in the pathogenesis of venous ulcers. We found that both microRNA (miR)-34a and miR-34c were upregulated in the wound-edge epidermal keratinocytes of venous ulcers compared with normal wounds or the skin. In keratinocytes, miR-34a and miR-34c promoted inflammatory chemokine and cytokine production. In wounds of wild-type mice, miR-34a–mimic treatment enhanced inflammation and delayed healing. To further explore how miR-34 functions, LGR4 was identified as a direct target mediating the proinflammatory function of miR-34a and miR-34c. Interestingly, impaired wound closure with enhanced inflammation was also observed in Lgr4 knockout mice. Mechanistically, the miR-34–LGR4 axis regulated GSK-3β–induced p65 serine 468 phosphorylation, changing the activity of the NF-κB signaling pathway. Collectively, the miR-34–LGR4 axis was shown to regulate keratinocyte inflammatory response, the deregulation of which may play a pathological role in venous ulcers.
Proline has been reported to play an important role in helping plants cope with several stresses, including salinity. This study investigates the relationship between proline accumulation and salt ...tolerance in an accession of Australian wild rice
Domin using morphological, physiological, and molecular assessments. Seedlings of
wild rice accession JC 2304 and two other cultivated rice
L. cultivars, Nipponbare (salt-sensitive), and Pokkali (salt-tolerant), were screened at 150 mM NaCl for 14 days. The results showed that
was able to rapidly accumulate free proline and lower osmotic potential at a very early stage of salt stress compared to cultivated rice. The qRT-PCR result revealed that
wild rice JC 2304 activated proline synthesis genes
and
and depressed the expression of proline degradation gene
as early as 1 h after exposure to salinity stress. Wild rice
and Pokkali maintained their relative water content and cell membrane integrity during exposure to salinity stress, while the salt-sensitive Nipponbare failed to do so. An analysis of the sodium and potassium contents suggested that
wild rice JC 2304 adapted to ionic stress caused by salinity by maintaining a low Na
content and low Na
/K
ratio in the shoots and roots. This demonstrates that
wild rice may use a rapid accumulation of free proline as a strategy to cope with salinity stress.
Objectives
The aim of this study was to investigate cementogenesis and alveolar bone induction during in vivo periodontal tissue regeneration upon implantation of hTGF‐β3 in furcation defects of ...Papio ursinus and to evaluate the feasibility of gene expression studies.
Materials and Methods
Class II furcation defects (day 0) were prepared in mandibular first and second molars of three P. ursinus and on day 30 implanted with and without 75 μg hTGF‐β3 in Matrigel®matrix. On day 0, 30 and 90, cementum and alveolar bone were harvested for gene expression analyses. Coral‐derived bioreactors with and without 250 μg hTGF‐β3 were implanted in the rectus abdominis to monitor tissue induction.
Results
hTGF‐β3 induced cementogenesis with TGF‐β3, Cementum Protein‐1 (Cemp1) and Osteocalcin (OC) up‐regulation, and down‐regulation of BMP‐2 and OP‐1. Matrigel®matrix specimens showed up‐regulation of BMP‐2, TGF‐β3, and OC, with down‐regulation of OP‐1 and Cemp1. hTGF‐β3 induced alveolar bone with down‐regulation of OP‐1, TGF‐β3, OC, and Cemp1. hTGF‐β3 bioreactors induced bone at the periphery only. BMP‐3, BMP‐4, TGF‐β1 and TGF‐β3 were up‐regulated in the adjacent muscle with TGF‐β2 down‐regulation.
Conclusions
Cementogenesis and osteogenesis by hTGF‐β3 entail the expression and up‐regulation of TGF‐β3 and OC with fine tuning and modulation of BMP‐2 and OP‐1.
This study aimed to examine the role of Artemisia annua in kidney functions in gentamicin-induced nephrotoxicity in mice.
In this study, 15 mice were used and divided into four groups. Each group has ...four mice; the first group is considered a control group with three mice due to receiving normal saline. Group II consists of an extract of Artemisia annua, group III consists of gentamicin, and Group IV consists of a combination of Artemisia annua and gentamicin. This process was continued for 15 days. All the mice were induced, and serum was extracted and used for biochemical parameters such as Creatinine, Urea, Uric acid, TNF-α, MDA, GSH, and Catalase (CAT) levels—additionally, histological and quantitative real-time PCR (qRT-PCR) analysis.
The results of this study confirmed biochemical values such as creatinine, Urea, and UA values showed a positive association (p<0.05), and showed a nominal association with histological analysis (p > 0.05). The Gentamicin group has a strong association with COX-2, NF-κB, and TGF-β genes (p < 0.05).
This study confirms gentamycin has a role in kidney functions with nephrotoxicity in mice and the protective effect of Artemisia annua.
•BES1 TFs regulate plant growth, development, and stress resistance, and play a pivotal role in the BR signal transduction pathway.•qRT-PCR analysis showed that the expression levels of VvBES1 genes ...differed in response to BR, MeJA, cold (-4 °C), NaCl, and PEG treatments.•VvBES1-3 overexpression enhances salt stress tolerance in transgenic Arabidopsis.•This study will contribute to further understanding the functions of BES1 TFs in the abiotic stress response.
BRI1-EMS-Suppressor 1 (BES1) regulates plant growth, development, and stress resistance, and plays a pivotal role in the brassinosteroid (BR) signal transduction pathway. In this study, a total of 12 BES1 genes were identified in the grape (Vitis vinifera) genome. Phylogenetic, structure, and motif sequence analyses of these genes provided insights into their evolutionary characteristics. Hormone-, stress-, and light-responsive and organ-specific cis-acting elements were identified in VvBES1 gene promoters. Microarray data analysis showed that VvBES1 family members exhibit diverse expression patterns in different organs. Quantitative real-time PCR (qRT-PCR) analysis showed that the expression levels of VvBES1 genes differed in response to BR, methyl jasmonate (MeJA), cold (4 °C), NaCl, and polyethylene glycol (PEG) treatments. The expression of VvBES1-3 was 29-fold higher under salt stress than control at 12 h. Moreover, VvBES1-3-overexpessing Arabidopsis thaliana plants showed lower malondialdehyde content, higher proline content, enhanced antioxidant enzyme (catalase, superoxide dismutase, peroxidase) activities, and higher salt-responsive gene expression levels than wild-type plants under salt stress, indicating that VvBES1-3 overexpression enhances salt stress tolerance in transgenic Arabidopsis. These results will contribute to further understanding the functions of BES1 transcription factors in the abiotic stress response.
► Astaxanthin accumulation by H. pluvialis was induced effectively by SA. ► All eight carotenogenic genes were up-regulated by SA treatments. ► Carotenogenic genes exhibited different mRNA expression ...profiles when exposed to SA. ► Surface of alga cells changed drastically along with astaxanthin accumulation.
The green alga Haematococcus pluvialis can produce large amounts of pink carotenoid astaxanthin which is a high value ketocarotenoid. In our study, transcriptional expression patterns of eight carotenoid genes in H. pluvialis in response to SA were measured using qRT-PCR. Results indicated that both 25 and 50mg/L salicylic acid (SA) could increase astaxanthin productivity and enhance transcriptional expression of eight carotenoid genes in H. pluvialis. But these genes exhibited different expression profiles. Moreover, SA25 (25mg/L SA) induction had a greater effect on the transcriptional expression of ipi-1, psy, pds, crtR-B and lyc (more than 6-fold up-regulation) than on ipi-2, bkt and crtO, but SA50 (50mg/L SA) treatment had a greater impact on the transcriptional expression of ipi-1, ipi-2, pds, crtR-B and lyc than on psy, bkt and crtO. Furthermore, astaxanthin biosynthesis under SA was up-regulated mainly by ipi-1, ipi-2, psy, crtR-B, bkt and crtO at transcriptional level, lyc at post-transcriptional level and pds at both levels. Summarily, these results suggest that SA constitute molecular signals in the network of astaxanthin biosynthesis. Induction of astaxanthin accumulation by SA without any other stimuli presents an attractive application potential in astaxanthin production with H. pluvialis.
Heavy metal contamination of water body has become a serious threat to aquatic life forms specially to fish. Hexavalent chromium (Cr VI) is one of the most potent heavy metal toxicant. It is present ...in aquatic environment at concentrations beyond permissible limit. Considering the fact that toxic effects are function of the exposure concentration, studies involving toxicological risk assessment should be done at environmentally relevant concentration. Therefore we studied the toxic effects of Cr VI to zebrafish at an environmentally relevant concentration (2 mg L−1). We monitored the genotoxic potential of Cr VI in erythrocytes through a simple reliable microscopic assay and found an increase in frequency of micronucleated erythrocytes along with erythrocytes with blebbed, lobed and notched nuclei. In addition, Cr VI induced neurotoxicity, being a least reported event was also investigated. Histological alterations in brain, elevated GSH and MDA content and increased catalase activity indicated oxidative stress-mediated damage. This was further confirmed through expressional alteration of Ucp2. Upregulation of Nrf2, Nqo1 and Ho1 clearly indicated the involvement of Nrf2-ARE system in stress response against Cr VI induced neurotoxicity. The transcriptional induction of apoptotic genes such as Bax, Caspase 9 and Caspase 3 along with downregulation of Bcl2 indicated that the cytoprotective system failed to counter the induced stress. Interestingly, there was upregulation of AChE gene, which could be correlated with the upregulated apoptotic genes. This study provides an insight on the neurotoxic stress of Cr VI on the zebrafish yet at an environmentally relevant concentration. Moreover the induction of nuclear anomalies in the erythrocytes can serve as extremely sensitive endpoints of toxicological stress indicators of aquatic contaminants like Cr VI.
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•Environmentally relevant concentration of Cr VI causes anomalies in zebrafish RBC.•2 mg L−1 Cr VI exposure caused neurotoxicity in zebrafish.•Involvement of Nrf2-ARE response system in Cr VI induced neurotoxicity.•Failure to cope up with stress results in upregulation of apoptotic genes.
The development and tissue-dependent color formation of the horticultural plant results in various color pattern flowers. Anthocyanins and carotenoids contribute to the red and yellow colors, ...respectively. In this study, quantitative real-time polymerase chain reaction (qRT-PCR) is used to analyze the expression profiles of anthocyanin and carotenoids biosynthesis genes in
(Rchb.f.) Rchb.f. Appropriate reference gene selection and validation are required before normalization of gene expression in qRT-PCR analysis. Thus, we firstly selected 12 candidate reference genes from transcriptome data, and used geNorm and Normfinder to evaluate their expression stability in lip (divided into abaxial and adaxial), petal, and sepal of the bud and flower of
. Our results show that the two most stable reference genes in different tissues of
bud and flower are
and
, the most unstable reference gene is
. The expression profiles of the
and
genes were similar to FPKM value profiles after normalization to the two most stable reference genes,
and
, with the upregulated
and
expression in flower stage, indicating that the ABP and CBP were activated across the stages of flower development. However, when the most unstable reference gene,
, was used to normalize the qRT-PCR data, the expression profiles of
and
differed from FPKM value profiles, indicating the necessity of selecting stable reference genes. Moreover,
and
expression was highest in the abaxial lip and adaxial lip, respectively, indicating that the ABP and CBP were activated in abaxial and adaxial lip, respectively, resulting in a presence of red or yellow segments in abaxial and adaxial lip. This study is the first to provide reference genes in
, and also provide useful information for studies that aim to understand the molecular mechanisms of flower color formation in
.
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